bims-meluca Biomed News
on Metabolism of non-small cell lung carcinoma
Issue of 2021‒12‒05
125 papers selected by
Cristina Muñoz Pinedo
L’Institut d’Investigació Biomèdica de Bellvitge
  1. Loss of DSTYK activates Wnt/β-catenin signaling and glycolysis in lung adenocarcinoma.
  2. The bioinformatics analysis of RIOX2 gene in lung adenocarcinoma and squamous cell carcinoma.
  3. Prognostic characterization of immune molecular subtypes in non-small cell lung cancer to immunotherapy.
  4. [Lung and gut microbiota and their interaction with the carcinogenesis and development of lung cancer: a review].
  5. Metabolomic Signature of Patients With Narcolepsy.
  6. Integrated DIA proteomics and lipidomics analysis on non-small cell lung cancer patients with TCM syndromes.
  7. Combining Metabolomics and Interpretable Machine Learning to Reveal Plasma Metabolic Profiling and Biological Correlates of Alcohol-Dependent Inpatients: What About Tryptophan Metabolism Regulation?
  8. Metabolic Reprogramming in the Tumor Microenvironment With Immunocytes and Immune Checkpoints.
  9. Prussian Blue/Calcium Peroxide Nanocomposites-Mediated Tumor Cell Iron Mineralization for Treatment of Experimental Lung Adenocarcinoma.
  10. Prognostic Implication of Direct Cardiac Invasion from Lung Cancer in Non-Operatively Treated Patients Based on Lung Computed Tomography Imaging.
  11. Genetic and epigenetic regulation of the NRF2-KEAP1 pathway in human lung cancer.
  12. Diversity and heterogeneity of immune states in non-small cell lung cancer and small cell lung cancer.
  13. Identification of the ferroptosis-related long non-coding RNAs signature to improve the prognosis prediction and immunotherapy response in patients with NSCLC.
  14. Derivation of a Novel CIHI in Patients with Lung Adenocarcinoma for Estimating Tumor Microenvironment and Clinical Prognosis.
  15. Subtyping non-small cell lung cancer by histology-guided spatial metabolomics.
  16. Survival Outcomes of Local Compared With Systemic First Treatment of Non-Small Cell Lung Cancer Brain Metastases.
  17. The impact of pathology grossing protocol measures to improve pathologic nodal staging in lung cancer.
  18. The association between gut microbiome affecting concomitant medication and the effectiveness of immunotherapy in patients with stage IV NSCLC.
  19. Amino acid deprivation induces AKT activation by inducing GCN2/ATF4/REDD1 axis.
  20. Functional assessment of somatic STK11 variants identified in primary human non-small cell lung cancers.
  21. The Prognostic Nutritional Index (PNI) is an independent predictor of overall survival in older patients with follicular lymphoma.
  22. Immune Checkpoint Inhibitors With or Without Bone-Targeted Therapy in NSCLC Patients With Bone Metastases and Prognostic Significance of Neutrophil-to-Lymphocyte Ratio.
  23. Decreased efficacy of the ketamine and scopolamine-induced sustained antidepressant-like effects in rats receiving metformin.
  24. Overall Survival and Safety With Pemetrexed/Platinum ± Anti-VEGF Followed by Pemetrexed ± Anti-VEGF Maintenance in Advanced Nonsquamous Non-Small-Cell Lung Cancer: A Pooled Analysis of 4 Randomized Studies.
  25. 1-deoxysphinganine initiates adaptive responses to serine and glycine starvation in cancer cells via proteolysis of sphingosine kinase.
  26. Downregulation of miR-29c promotes muscle wasting by modulating the activity of leukemia inhibitory factor in lung cancer cachexia.
  27. Testis developmental related gene 1 promotes non-small-cell lung cancer through the microRNA-214-5p/Krüppel-like factor 5 axis.
  28. Seven autophagy-related lncRNAs are associated with the tumor immune microenvironment in predicting survival risk of nonsmall cell lung cancer.
  29. Circular RNA circVAPA contributes to non-small-cell lung cancer progression via miR-342-3p-dependent regulation of ZEB2.
  30. Targeting HSPA1A in ARID2-deficient lung adenocarcinoma.
  31. The effects of naringenin and naringin on the glucose uptake and AMPK phosphorylation in high glucose treated HepG2 cells.
  32. Beyond epidermal growth factor receptor: MET amplification as a general resistance driver to targeted therapy in oncogene-driven non-small-cell lung cancer.
  33. Correlation of serum electrolytes with serial miRNA in advanced stage non-small cell lung cancer (NSCLC) in Indonesia.
  34. Enhancing the Therapeutic Efficacy of KRASG12C Inhibitors in Lung Adenocarcinoma Cell Models by Cotargeting the MAPK Pathway or HSP90.
  35. LncRNA ADAMTS9-AS2 is a Prognostic Biomarker and Correlated with Immune Infiltrates in Lung Adenocarcinoma.
  36. VLX1570 regulates the proliferation and apoptosis of human lung cancer cells through modulating ER stress and the AKT pathway.
  37. Effects of CT images at different reconstruction energies on radiotherapy planning of patients diagnosed with nonsmall cell lung cancer.
  38. Methods of nutritional assessment and functional capacity in the identification of unfavorable clinical outcomes in hospitalized patients with cancer: a systematic review.
  39. Inhibitory effects of icotinib combined with antiangiogenic drugs in human non-small cell lung cancer xenograft models are better than single target drugs.
  40. Metformin Suppresses Monocyte Immunometabolic Activation by SARS-CoV-2 Spike Protein Subunit 1.
  41. CircRPPH1 promotes cell proliferation, migration and invasion of non-small cell lung cancer (NSCLC) via the PI3K/AKT and JAK2/STAT3 signaling axes.
  42. The prevalence of CT-defined low skeletal muscle mass in patients with metastatic cancer: a cross-sectional multicenter French study (the SCAN study).
  43. Countering the advert effects of lung cancer on the anticancer potential of dendritic cell populations reinstates sensitivity to anti-PD-1 therapy.
  44. Pharmacokinetics, Mass Balance, and Metabolism of the Novel URAT1 Inhibitor [14C]HR011303 in Humans: Metabolism is Mediated Predominantly by UDP-glucuronosyltransferase.
  45. Deubiquitinase USP5 promotes non-small cell lung cancer cell proliferation by stabilizing cyclin D1.
  46. Gene expression signatures as candidate biomarkers of response to PD-1 blockade in non-small cell lung cancers.
  47. Distinct properties of pure- and mixed-type high-grade fetal lung adenocarcinomas by genetic profiling and transcription factor expression.
  48. Efficacy and safety of MIL60 compared with bevacizumab in advanced or recurrent non-squamous non-small cell lung cancer: a phase 3 randomized, double-blind study.
  49. PESV represses non-small cell lung cancer cell malignancy through circ_0016760 under hypoxia.
  50. Targeting the metabolism of cancer stem cells by energy disruptor molecules.
  51. Gut microbiome signatures distinguish type 2 diabetes mellitus from non-alcoholic fatty liver disease.
  52. Plasma SNORD83A as a potential biomarker for early diagnosis of non-small-cell lung cancer.
  53. Aberrant Alteration of Circulating Lymphocyte Subsets in Small Cell Lung Cancer Patients Treated with Radiotherapy.
  54. Tumour-infiltrating bystander CD8+ T cells activated by IL-15 contribute to tumour control in non-small cell lung cancer.
  55. Prevalence and prognostic significance of malnutrition in diabetic patients with coronary artery disease: a cohort study.
  56. Keratinocyte differentiation antigen-specific T cells in immune checkpoint inhibitor-treated NSCLC patients are associated with improved survival.
  57. BLM helicase inhibition synergizes with PARP inhibition to improve the radiosensitivity of olaparib resistant non-small cell lung cancer cells by inhibiting homologous recombination repair.
  58. CircRNA circ_0008037 facilitates tumor growth and the Warburg effect via upregulating NUCKS1 by binding to miR-433-3p in non-small cell lung cancer.
  59. A new ALK inhibitor overcomes resistance to first- and second-generation inhibitors in NSCLC.
  60. Prognostic significance of postoperative longitudinal change of serum carcinoembryonic antigen level in patients with stage I lung adenocarcinoma completely resected by single-port video-assisted thoracic surgery: a retrospective study.
  61. Metabolomic signatures after bariatric surgery - a systematic review.
  62. Metabolomic analysis and oxidative stress response reveals the toxicity in Escherichia coli induced by organophosphate flame retardants tris(2-chloroethyl) phosphate and triphenyl phosphate.
  63. Baseline tumour size is an independent prognostic factor for overall survival in PD-L1 ≥ 50% non-small cell lung cancer patients treated with first-line pembrolizumab.
  64. MALAT-1 Expression Correlates with Prognosis in Non-Small-Cell Lung Carcinoma: A Systematic Review and Meta-analysis.
  65. Impact of muscle mass on blood glucose level.
  66. Progesterone Receptor Membrane Component 1 and Its Accomplice: Emerging Therapeutic Targets in Lung Cancer.
  67. LncRNA PKMYT1AR promotes cancer stem cell maintenance in non-small cell lung cancer via activating Wnt signaling pathway.
  68. Molecular Characterization of the Clinical and Tumor Immune Microenvironment Signature of 5-methylcytosine-Related Regulators in non-small Cell Lung Cancer.
  69. Association of PTPRT mutations with immune checkpoint inhibitors response and outcome in melanoma and non-small cell lung cancer.
  70. Oxidative stress parameters and keap 1 variants in T2DM: Association with T2DM, diabetic neuropathy, diabetic retinopathy, and obesity.
  71. Intradialytic nutrition and quality of life in Chilean older patients in hemodialysis with protein-energy wasting.
  72. Menstrual factors, reproductive history, and risk of lung cancer: a multi-center population-based cohort study in Chinese females.
  73. Failure to EGFR-TKI-based therapy and tumoural progression are promoted by MEOX2/GLI1-mediated epigenetic regulation of EGFR in the human lung cancer.
  74. WSF-CT-11, a Sesquiterpene Derivative, Activates AMP-Activated Protein Kinase with Anti-diabetic Effects in 3T3-L1 Adipocytes.
  75. Comparison of prophylactic percutaneous endoscopic gastrostomy with reactive enteral nutrition in patients with head and neck cancer undergoing radiotherapy or chemoradiotherapy: A systematic review.
  76. Targeting cancer metabolism in the era of precision oncology.
  77. Development and validation of a Modified Patient-Generated Subjective Global Assessment as a nutritional assessment tool in cancer patients.
  78. Construction and validation of a metabolic gene-associated prognostic model for cervical carcinoma and the role on tumor microenvironment and immunity.
  79. Ghrelin is a signal to facilitate the utilization of fatty acids and save glucose by the liver, skeletal muscle, and adipose tissues in chicks.
  80. Altered Oxidative Status as a Cost of Reproduction in a Seabird with High Reproductive Costs.
  81. Lung Cancer Screening.
  82. Metabolic Profile in Plasma AND CSF of LEVODOPA-induced Dyskinesia in Parkinson's Disease: Focus on Neuroinflammation.
  83. Cysteine 253 of UCP1 regulates energy expenditure and sex-dependent adipose tissue inflammation.
  84. Comparison of Beinaglutide Versus Metformin for Weight Loss in Overweight and Obese Non-diabetic Patients.
  85. Anti-obesity effect of Auricularia heimuer fruiting body alcohol extraction on obese mice and crucial metabolite pathway analysis by liquid chromatography-tandem mass spectrometry.
  86. Post-Progression Survival Highly Influences Overall Survival in Driver Gene Mutation/Translocation Negative or Unknown Type of Non-Small Cell Lung Cancer.
  87. Are there differences by sex in lung cancer characteristics at diagnosis? -a nationwide study.
  88. Therapeutic targeting of SPIB/SPI1-facilitated interplay of cancer cells and neutrophils inhibits aerobic glycolysis and cancer progression.
  89. Analysis at the single-cell level indicates an important role of heterogeneous global DNA methylation status on the progression of lung adenocarcinoma.
  90. Impact of Tumor Suppressor Gene Co-Mutations on Differential Response to EGFR TKI Therapy in EGFR L858R and Exon 19 Deletion Lung Cancer.
  91. Clinical validation of Guardant360 CDx as a blood-based companion diagnostic for sotorasib.
  92. Immunotherapy for cancer: effects of iron oxide nanoparticles on polarization of tumor-associated macrophages.
  93. Fluvastatin Reduces Glucose Tolerance in Healthy Young Individuals Independently of Cold Induced BAT Activity.
  94. Quality of life, HPV-status and phase angle predict survival in head and neck cancer patients under (chemo)radiotherapy undergoing nutritional intervention: Results from the prospective randomized HEADNUT-trial.
  95. Identification and Quantification of MIDD0301 metabolites.
  96. CT pectoralis muscle area is associated with DXA lean mass and correlates with emphysema progression in a tobacco-exposed cohort.
  97. Oncogenic dependency on STAT3 serine phosphorylation in KRAS mutant lung cancer.
  98. Uncovering the Metabolic Mechanism of Salidroside Alleviating Microglial Hypoxia Inflammation Based on Microfluidic Chip-Mass Spectrometry.
  99. Metformin anticipates peak of lactate during high-intensity interval training but no changes performance or neuromuscular response in amateur swimmers.
  100. Use of oral glutamine in radiation-induced adverse effects in patients with thoracic and upper aerodigestive malignancies: Results of a prospective observational study.
  101. Mutant-IDH inhibits Interferon-TET2 signaling to promote immunoevasion and tumor maintenance in cholangiocarcinoma.
  102. Baicalein resensitizes tamoxifen-resistant breast cancer cells by reducing aerobic glycolysis and reversing mitochondrial dysfunction via inhibition of hypoxia-inducible factor-1α.
  103. Computed tomography-guided cryoablation for adrenal metastases secondary to lung cancer.
  104. FeV1 and BMI influence King's Sarcoidosis Questionnaire score in sarcoidosis patients.
  105. NEDD4L-induced ubiquitination mediating UBE2T degradation inhibits progression of lung adenocarcinoma via PI3K-AKT signaling.
  106. SLC25A1 promotes tumor growth and survival by reprogramming energy metabolism in colorectal cancer.
  107. Probiotic supplementation improves clinical outcomes and quality of life indicators in patients with plaque psoriasis: A randomized double-blind clinical trial.
  108. Fermentation supernatant of Staphylococcus aureus drives catabolism in chondrocytes via NF-κB signaling mediated increase of cholesterol metabolism.
  109. Nutritional and prognostic significance of abdominal wall thickness measured during percutaneous endoscopic gastrostomy in older individuals with dysphagia.
  110. L'impatto dell'esposizione occupazionale ad amianto sul tumore del polmone in Italia.
  111. Roles of hydroxyacyl-CoA dehydrogenase trifunctional multienzyme complex subunit alpha, a lipid metabolism enzyme, in Wilms tumor patients.
  112. Human Rhinovirus Infection of the Respiratory Tract Affects Sphingolipid Synthesis.
  113. Lower Prevalence of Osteoporosis in Patients with COPD Taking Anti-Inflammatory Compounds for the Treatment of Diabetes: Results from COSYCONET.
  114. Profound neutropenia related to consolidation durvalumab following concurrent radiochemotherapy in III stage non-small-cell lung cancer patient.
  115. [Advances in research on the relationship between changes in cellular metabolism and lung diseases].
  116. Lung cancer for the non-respiratory physician.
  117. Synergistic alterations in the multilevel chromatin structure anchor dysregulated genes in small cell lung cancer.
  118. LncRNA SFTA1P mediates positive feedback regulation of the Hippo-YAP/TAZ signaling pathway in non-small cell lung cancer.
  119. [Research Progress in Glucose Metabolism of Chondrocytes].
  120. Pharmacokinetic and pharmacogenomic analysis of low-dose afatinib treatment in elderly patients with EGFR mutation-positive non-small cell lung cancer.
  121. Preoperative Prognostic Nutritional Index is a significant predictive factor for posthepatectomy bile leakage.
  122. Comprehensive analysis of PD-L1 in non-small cell lung cancer with emphasis on survival benefit, impact of driver mutation and histological types, and archival tissue.
  123. Evaluation of Clinical Performance of TiNi-Based Implants Used in Chest Wall Repair after Resection for Malignant Tumors.
  124. Simultaneous Evaluations of pH and Enzyme Activity with a CEST MRI Contrast Agent.
  125. Afatinib mediates autophagic degradation of ORAI1, STIM1, and SERCA2, which inhibits proliferation of non-small cell lung cancer cells.
  1. Cell Death Dis. 2021 Dec 01. 12(12): 1122
    Loss of DSTYK activates Wnt/β-catenin signaling and glycolysis in lung adenocarcinoma.
    Chenxi Zhong, Ming Chen, Yu Chen, Feng Yao, Wentao Fang.
      Aberrant activation of Wnt/β-catenin signaling and dysregulation of metabolism have been frequently observed in lung cancer. However, the molecular mechanism by which Wnt/β-catenin signaling is regulated and the link between Wnt/β-catenin signaling and cancer metabolism are not fully understood. In this study, we showed that the loss of dual serine/threonine tyrosine protein kinase (DSTYK) led to the activation of Wnt/β-catenin signaling and upregulation of its target gene, lactate dehydrogenase (LDHA), and thus the elevation of lactate. DSTYK phosphorylated the N-terminal domain of β-catenin and inhibited Wnt/β-catenin signaling, which led to the inhibition of cell growth, colony formation and tumorigenesis in a lung adenocarcinoma mouse model. DSTYK was downregulated in lung cancer tissues, and its expression was positively correlated with the survival of patients with lung adenocarcinoma. Taken together, these results demonstrate that the loss of DSTYK activates Wnt/β-catenin/LDHA signaling to promote the tumorigenesis of lung cancer and that DSTYK may be a therapeutic target.
    DOI:  https://doi.org/10.1038/s41419-021-04385-1
  2. PLoS One. 2021 ;16(12): e0259447
    The bioinformatics analysis of RIOX2 gene in lung adenocarcinoma and squamous cell carcinoma.
    Bingqing Sun, Hongwen Zhao.
      Lung cancer is characterized by high morbidity and mortality rates, and it has become an important public health issue worldwide. The occurrence and development of tumors is a multi-gene and multi-stage complex process. As an oncogene, ribosomal oxygenase 2 (RIOX2) has been associated with a variety of cancers. In this article, we analyzed the correlation between RIOX2 expression and methylation in lung cancer based on the databases including the cancer genome atlas (TCGA) (https://portal.gdc.cancer.gov/) and the gene expression omnibus (GEO) (https://www.ncbi.nlm.nih.gov/geo/). It was found that RIOX2 is highly expressed in lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC) tissues, whose expression is negatively correlated with its methylation level. In this regard, methylation at cg09716038, cg14773523, cg14941179, and cg22299097 had a significant negative correlation with RIOX2 expression in LUAD, whereas in LUSC, methylation at cg09716038, cg14773523, cg14941179, cg22299097, cg05451573, cg10779801, and cg23629183 is negatively correlated with RIOX2 expression. According to the analysis based on the databases, RIOX2 gene could not be considered as the independent prognostic biomarker in lung adenocarcinoma or squamous cell lung cancer. However, the molecular mechanism of RIOX2 gene in the development of lung cancer may be helpful in improving lung cancer therapy.
    DOI:  https://doi.org/10.1371/journal.pone.0259447
  3. BMC Pulm Med. 2021 Nov 29. 21(1): 389
    Prognostic characterization of immune molecular subtypes in non-small cell lung cancer to immunotherapy.
    Chenlu Li, Jingjing Pan, Jing Luo, Xupeng Chen.
      BACKGROUND: Non-small cell lung cancer (NSCLC) was usually associated with poor prognosis and invalid therapeutical response to immunotherapy due to biological heterogeneity. It is urgent to screen reliable biomarkers, especially immunotherapy-associated biomarkers, that can predict outcomes of these patients.METHODS: Gene expression profiles of 1026 NSCLC patients were collected from The Cancer Genome Atlas (TCGA) datasets with their corresponding clinical and somatic mutation data. Based on immune infiltration scores, molecular clustering classification was performed to identify immune subtypes in NSCLC. After the functional enrichment analysis of subtypes, hub genes were further screened using univariate Cox, Lasso, and multivariate Cox regression analysis, and the risk score was defined to construct the prognostic model. Other microarray data and corresponding clinical information of 603 NSCLC patients from the GEO datasets were applied to conduct random forest models for the prognosis of NSCLC with 100 runs of cross-validation. Finally, external datasets with immunotherapy and chemotherapy were further applied to explore the significance of risk-scores in clinical immunotherapy response for NSCLC patients.
    RESULTS: Compared with Subtype-B, the Subtype-A, associated with better outcomes, was characterized by significantly higher stromal and immune scores, T lymphocytes infiltration scores and up-regulation of immunotherapy markers. In addition, we found and validated an eleven -gene signatures for better application of distinguishing high- and low-risk NSCLC patients and predict patients' prognosis and therapeutical response to immunotherapy. Furthermore, combined with other clinical characteristics based on multivariate Cox regression analysis, we successfully constructed and validated a nomogram to effectively predict the survival rate of NSCLC patients. External immunotherapy and chemotherapy cohorts validated the patients with higher risk-scores exhibited significant therapeutic response and clinical benefits.
    CONCLUSION: These results demonstrated the immunological and prognostic heterogeneity within NSCLC and provided a new clinical application in predicting the prognosis and benefits of immunotherapy for the disease.
    Keywords:  Immune molecular subtype; Immunotherapy; Non-small cell lung cancer; Prognosis; Risk stratification
    DOI:  https://doi.org/10.1186/s12890-021-01765-3
  4. Sheng Wu Gong Cheng Xue Bao. 2021 Nov 25. 37(11): 3789-3800
    [Lung and gut microbiota and their interaction with the carcinogenesis and development of lung cancer: a review].
    Yaokun Zhang, Youming Zhang, Hongli Si.
      Lung microbiota and gut microbiota are closely related to lung cancer. Studies have shown that the dysbiosis, i.e., the significantly altered composition and structure of gut and lung microbiota, usually occurs in patients with lung cancer. With the introduction of "Gut-Lung Axis", an increasing attention has been paid to the close relationship between the lung and gut microbiota in human body. A deeper insight into this relationship would facilitate understanding the mechanisms behind the carcinogenesis and development of lung cancer. This article summarizes the composition of lung and gut microbiota in patients with lung cancer and the possible interaction mechanisms, highlighting the importance of the immune system in the Gut-Lung Axis. The effects of lung and gut microbiota on the clinical treatment of lung cancer were summarized, based on which the authors propose that the lung and gut microbiota can be used as novel targets for early diagnosis and treatment of lung cancer.
    Keywords:  gut microbiota; gut-lung axis; immune & therapy; lung cancer; lung microbiota
    DOI:  https://doi.org/10.13345/j.cjb.210081
  5. Neurology. 2021 Nov 29. pii: 10.1212/WNL.0000000000013128. [Epub ahead of print]
    Metabolomic Signature of Patients With Narcolepsy.
    Dauvilliers Yves, Lucie Barateau, Benita Middleton, Daan Van Der Veen, Debra J Skene.
      BACKGROUND AND OBJECTIVE: Narcolepsy type 1 (NT1) is an orphan brain disorder caused by the irreversible destruction of orexin neurons. Metabolic disturbances are common in patients with NT1 who have a body mass index (BMI) 10-20% higher than the general population, with one third being obese (BMI>30 kg/m2). Besides the destruction of orexin neurons in NT1, the metabolic alterations in obese and non-obese patients with narcolepsy type 1 remain unknown. The aim of the study was to identify possible differences in plasma metabolic profiles between patients with NT1 and controls as a function of their BMI status.METHODS: We used a targeted liquid chromatography-mass spectrometry metabolomics approach to measure 141 circulating, low molecular weight metabolites in drug-free fasted plasma samples from 117 NT1 patients (including 41 obese subjects) compared with 116 BMI-matched controls (including 57 obese subjects).
    RESULTS: Common metabolites driving the difference between NT1 and controls, irrespective of BMI, were identified, namely sarcosine, glutamate, nonaylcarnitine (C9), 5 long chain lysophosphatidylcholine acyls, one sphingolipid, 12 phosphatidylcholine diacyls and 11 phosphatidylcholine acyl-akyls, all showing increased concentrations in NT1. Metabolite concentrations significantly affected by NT1 (n = 42) and BMI category (n = 40) showed little overlap (n = 5). Quantitative enrichment analysis revealed common metabolic pathways that were implicated in the NT1/control differences, in both normal BMI and obese comparisons, namely glycine and serine, arachidonic acid, and tryptophan metabolisms. The metabolites driving these differences were glutamate, sarcosine and ornithine (glycine and serine metabolism), glutamate and PC aa C34:4 (arachidonic acid metabolism) and glutamate, serotonin and tryptophan (tryptophan metabolism). Linear metabolite-endophenotype regression analyses highlight that as part of the NT1 metabolic phenotype, most of the relationships between the sleep parameters (i.e. slow wave sleep duration, sleep latency and periodic leg movement) and metabolite concentrations seen in the controls were lost.
    DISCUSSION: These results represented the most comprehensive metabolic profiling of patients with NT1 as a function of BMI and propose some metabolic diagnostic biomarkers for NT1, namely glutamate, sarcosine, serotonin, tryptophan, nonaylcarnitine and some phosphatidylcholines. The metabolic pathways identified offer, if confirmed, possible targets for treatment of obesity in NT1.
    CLASSIFICATION OF EVIDENCE: This study provides Class II evidence that a distinct metabolic profile can differentiate patients with Narcolepsy Type 1 from patients without the disorder.
    DOI:  https://doi.org/10.1212/WNL.0000000000013128
  6. Chin Med. 2021 Nov 27. 16(1): 126
    Integrated DIA proteomics and lipidomics analysis on non-small cell lung cancer patients with TCM syndromes.
    Song Cang, Ran Liu, Wei Jin, Qi Tang, Wanjun Li, Kunqian Mu, Pengfei Jin, Kaishun Bi, Qing Li.
      BACKGROUND: Lung cancer remains the leading cause of mortality from malignant tumors, non-small cell lung cancer (NSCLC) accounts for the majority of lung cancer cases, and individualized diagnosis and treatment is an effective trend. The individual characteristics of different traditional Chinese medicine (TCM) syndromes of NSCLC patients may be revealed by highly specific molecular profiles.METHODS: In this study, 10 NSCLC patients with Qi deficiency and Yin deficiency (QDYD) syndrome and 10 patients with Qi deficiency of lung-spleen (QDLS) syndrome in TNM stage III-IV as well as 10 healthy volunteers were enrolled. Aiming at the varied syndromes of NSCLC patients with "Yin deficiency" as the main difference, a proteomics research based on data-independent acquisition (DIA) was developed. Of the dysregulated proteins in NSCLC patients, lipid metabolism was significantly enriched. Thereafter, nontargeted lipidomics research based on UPLC-Q-TOF/MS was performed in 16 patients, with 8 individuals randomly selected from each syndrome group. Furthermore, the considerably different characteristics between the syndromes and pathological mechanisms of NSCLC were screened by statistical and biological integrations of proteomics and lipidomics and the differential metabolic pathways of the two similar syndromes were further explored. Besides, lipids biomarkers were verified by a clinically used anticancer Chinese medicine, and the level of key differential proteins in the two syndromes was also validated using ELISA.
    RESULTS: The results showed that glycerophospholipid metabolism, sphingolipid metabolism, glycolipid metabolism, and primary bile acid biosynthesis were altered in NSCLC patients and that glycerophospholipid metabolism was significantly changed between the two syndromes in lipidomics analysis. Among the proteins and lipids, ALDOC and lysophosphatidylcholine (LPCs) were revealed to have a strong relationship by statistical and biological integration analysis, and could effectively distinguish QDLS and QDYD syndromes. Notably, the patients with different syndromes had the most typical metabolic patterns in glycerophospholipid metabolism and glycolysis, reflecting the differences in the syndromes dominated by "Yin deficiency".
    CONCLUSIONS: ALDOC and LPCs could be employed for the differentiation of NSCLC patients with QDLS and QDYD syndromes, and "Yin deficiency" might be associated with glycerophospholipid metabolism and glycolysis pathway. The results provided a theoretical basis for "Syndrome differentiation" in TCM diagnosis. Moreover, the developed integrated strategy could also provide a reference for individualized diagnosis and treatment of other diseases.
    Keywords:  Lipidomics; Non-small cell lung cancer; Proteomics; Syndrome differentiation; Traditional Chinese medicine syndromes
    DOI:  https://doi.org/10.1186/s13020-021-00535-x
  7. Front Mol Biosci. 2021 ;8 760669
    Combining Metabolomics and Interpretable Machine Learning to Reveal Plasma Metabolic Profiling and Biological Correlates of Alcohol-Dependent Inpatients: What About Tryptophan Metabolism Regulation?
    Xiuqing Zhu, Jiaxin Huang, Shanqing Huang, Yuguan Wen, Xiaochang Lan, Xipei Wang, Chuanli Lu, Zhanzhang Wang, Ni Fan, Dewei Shang.
      Alcohol dependence (AD) is a condition of alcohol use disorder in which the drinkers frequently develop emotional symptoms associated with a continuous alcohol intake. AD characterized by metabolic disturbances can be quantitatively analyzed by metabolomics to identify the alterations in metabolic pathways. This study aimed to: i) compare the plasma metabolic profiling between healthy and AD-diagnosed individuals to reveal the altered metabolic profiles in AD, and ii) identify potential biological correlates of alcohol-dependent inpatients based on metabolomics and interpretable machine learning. Plasma samples were obtained from healthy (n = 42) and AD-diagnosed individuals (n = 43). The plasma metabolic differences between them were investigated using liquid chromatography-tandem mass spectrometry (AB SCIEX® QTRAP 4500 system) in different electrospray ionization modes with scheduled multiple reaction monitoring scans. In total, 59 and 52 compounds were semi-quantitatively measured in positive and negative ionization modes, respectively. In addition, 39 metabolites were identified as important variables to contribute to the classifications using an orthogonal partial least squares-discriminant analysis (OPLS-DA) (VIP > 1) and also significantly different between healthy and AD-diagnosed individuals using univariate analysis (p-value < 0.05 and false discovery rate < 0.05). Among the identified metabolites, indole-3-carboxylic acid, quinolinic acid, hydroxy-tryptophan, and serotonin were involved in the tryptophan metabolism along the indole, kynurenine, and serotonin pathways. Metabolic pathway analysis revealed significant changes or imbalances in alanine, aspartate, glutamate metabolism, which was possibly the main altered pathway related to AD. Tryptophan metabolism interactively influenced other metabolic pathways, such as nicotinate and nicotinamide metabolism. Furthermore, among the OPLS-DA-identified metabolites, normetanephrine and ascorbic acid were demonstrated as suitable biological correlates of AD inpatients from our model using an interpretable, supervised decision tree classifier algorithm. These findings indicate that the discriminatory metabolic profiles between healthy and AD-diagnosed individuals may benefit researchers in illustrating the underlying molecular mechanisms of AD. This study also highlights the approach of combining metabolomics and interpretable machine learning as a valuable tool to uncover potential biological correlates. Future studies should focus on the global analysis of the possible roles of these differential metabolites and disordered metabolic pathways in the pathophysiology of AD.
    Keywords:  alcohol dependence; biological correlate; machine learning; metabolic pathway; metabolic profiling; metabolomics; orthogonal partial least squares-discriminant analysis; tryptophan metabolism
    DOI:  https://doi.org/10.3389/fmolb.2021.760669
  8. Front Oncol. 2021 ;11 759015
    Metabolic Reprogramming in the Tumor Microenvironment With Immunocytes and Immune Checkpoints.
    Yaolin Xu, Lijie He, Qiang Fu, Junzhe Hu.
      Immune checkpoint inhibitors (ICIs), Ipilimumab, Nivolumab, Pembrolizumab and Atezolizumab, have been applied in anti-tumor therapy and demonstrated exciting performance compared to conventional treatments. However, the unsatisfactory response rates, high recurrence and adaptive resistance limit their benefits. Metabolic reprogramming appears to be one of the crucial barriers to immunotherapy. The deprivation of required nutrients and altered metabolites not only promote tumor progression but also confer dysfunction on immune cells in the tumor microenvironment (TME). Glycolysis plays a central role in metabolic reprogramming and immunoregulation in the TME, and many therapies targeting glycolysis have been developed, and their combinations with ICIs are in preclinical and clinical trials. Additional attention has been paid to the role of amino acids, lipids, nucleotides and mitochondrial biogenesis in metabolic reprogramming and clinical anti-tumor therapy. This review attempts to describe reprogramming metabolisms within tumor cells and immune cells, from the aspects of glycolysis, amino acid metabolism, lipid metabolism, nucleotide metabolism and mitochondrial biogenesis and their impact on immunity in the TME, as well as the significance of targeting metabolism in anti-tumor therapy, especially in combination with ICIs. In particular, we highlight the expression mechanism of programmed cell death (ligand) 1 [PD-(L)1] in tumor cells and immune cells under reprogramming metabolism, and discuss in detail the potential of targeting key metabolic pathways to break resistance and improve the efficacy of ICIs based on results from current preclinical and clinical trials. Besides, we draw out biomarkers of potential predictive value in ICIs treatment from a metabolic perspective.
    Keywords:  PD-1; amino acid metabolism; glycolysis; immune checkpoints; lipid metabolism; mitochondrial biogenesis; nucleotide metabolism; the tumor microenvironment
    DOI:  https://doi.org/10.3389/fonc.2021.759015
  9. ACS Nano. 2021 Dec 01.
    Prussian Blue/Calcium Peroxide Nanocomposites-Mediated Tumor Cell Iron Mineralization for Treatment of Experimental Lung Adenocarcinoma.
    Kaixin Zhang, Jicheng Wu, Xiaoxiong Zhao, Jiale Qin, Yi Xue, Weizeng Zheng, Lixiu Wang, Haoran Wang, Hong Shen, Tianye Niu, Yan Luo, Ruikang Tang, Ben Wang.
      Current lung cancer diagnosis methods encounter delayed visual confirmation of tumor foci and low-resolution metrics in imaging findings, which delays the early treatment of tumors. Here, we developed a potent lung cancer imaging and treatment strategy centered around a nanotransformational concept of tumor iron mineralization in situ, which employs Prussian blue/calcium peroxide nanocomposites as a precursor. The resultant iron mineralization in tumor cells greatly facilitates the early and differential diagnosis of lung carcinoma from benign nodules via medical imaging, meanwhile introducing oxidative stress to activate the cellular apoptosis and ferroptosis pathways, resulting in inhibition of the malignant behavior of tumor cells. Tumor-microenvironment-triggered iron mineralization enables integration of the detection and prevention of tumor metastasis at its early stages with no assistance of toxic drugs, which offers a potential solution for the precise management of lung cancer with ideal outcomes.
    Keywords:  antitumor therapy; biomineralization; differential diagnosis; iron mineralization; lung cancer
    DOI:  https://doi.org/10.1021/acsnano.1c07308
  10. Heart Lung Circ. 2021 Nov 25. pii: S1443-9506(21)01315-9. [Epub ahead of print]
    Prognostic Implication of Direct Cardiac Invasion from Lung Cancer in Non-Operatively Treated Patients Based on Lung Computed Tomography Imaging.
    Ying Zhong, Ce Li, Yuehuan Sheng, Jiaqi Wang, Guan Wang.
      BACKGROUND: Lung cancer with direct cardiac invasion (LCCI+) exerts a significant influence on the survival of patients. There is a paucity of comparative research into the prognosis of advanced lung cancer with and without direct cardiac invasion.METHOD: In this study, 50 LCCI+ patients and 50 sex-, age-, and TNM stage-matched patients without direct cardiac invasion (LCCI-) were retrospectively analysed. LCCI+ was defined as lung cancer directly invading the heart by penetrating mediastinum or extending into the atrium via the pulmonary vein. The study endpoint was all-cause death. In this study, the survival time was defined as the time from the first detection of direct cardiac invasion to the end of the event.
    RESULTS: During a median follow-up period of 31 months, all-cause death occurred in 44 patients (88.0%) in the LCCI+ group and in 36 patients (72.0%) in the LCCI- group; the overall survival (OS) time among patients in the LCCI+ group was significantly lower compared with those in the LCCI- group (5.0 [interquartile range (IQR), 2.0-12.0] vs 13.8 [IQR, 4.0-18.4] months; p<0.001); the OS rate in the LCCI+ group was significantly lower compared with patients in the LCCI- group (log-rank, p=0.0002). Multivariate Cox regression analysis showed that direct cardiac invasion was an independent predictor of survival in patients with advanced lung cancer (hazard ratio, 2.255; 95% confidence interval, 1.443-3.524). Further analysis indicated that in patients with small cell lung cancer, the survival rate between the LCCI+ group and LCCI- group was insignificant (log-rank, p=0.075; survival time: 4.0 [IQR, 2.0-11.5] vs 11.5 [IQR, 5.0-18.3] months); in patients with non-small cell lung cancer (NSCLC), the survival rate in the LCCI+ group was lower than that of the LCCI- group (log-rank, p=0.01; survival time: 6.0 [IQR, 3.0-13.3] vs 16.3 [IQR, 10.4-27.2] months).
    CONCLUSIONS: Direct cardiac invasion from lung cancer was an independent prognostic factor for survival time in patients with lung cancer. Patients with direct cardiac invasion by NSCLC have a poorer clinical outcome than those without direct cardiac invasion. A careful preoperative evaluation is mandatory and appropriate management of cardiac involvement should be considered in the treatment of NSCLC.
    Keywords:  Advanced lung cancer; Cardiac metastases; Cardio-oncology; Prognosis
    DOI:  https://doi.org/10.1016/j.hlc.2021.10.018
  11. Br J Cancer. 2021 Nov 29.
    Genetic and epigenetic regulation of the NRF2-KEAP1 pathway in human lung cancer.
    Nuria Camiña, Trevor M Penning.
      Electrophilic and oxidative stress is caused when homeostatic mechanisms are disrupted. A major defense mechanism involves the activation of the nuclear factor erythroid 2-related factor 2 (NRF2) transcription factor encoded by the NFE2L2 gene, which can accelerate the detoxification of electrophilic carcinogens and prevent cancer and on the other hand in certain exposure contexts may exacerbate the carcinogenic process. NRF2-target genes activated under these conditions can be used as biomarkers of stress signalling, while activation of NRF2 can also reveal the epigenetic mechanisms that modulate NFE2L2 expression. Epigenetic mechanisms that regulate NFE2L2 and the gene for its adaptor protein KEAP1 include DNA methylation, histone modifications and microRNA. Understanding the activation of the NRF2-KEAP1 signalling pathway in human lung cancer, its epigenetic regulation and its role in oncogenesis is the subject of this review.
    DOI:  https://doi.org/10.1038/s41416-021-01642-0
  12. PLoS One. 2021 ;16(12): e0260988
    Diversity and heterogeneity of immune states in non-small cell lung cancer and small cell lung cancer.
    Shawn J Rice, Chandra P Belani.
      Blood-based biomarkers including systemic inflammation (SI) indicators or circulating factors (cytokines, chemokines, or growth factors) are associated with a poor prognosis for lung cancer patients. Collectively these biomarkers can predict the immune state of a patient. We wanted to define and compare the immune states of small cell and non-small cell lung cancer patients, in the hopes that the information gained could lead to overall improvements in patient care and outcomes. Specimens and data from 235 patients was utilized, 49 surgically resected non-small cell lung cancer (NSCLC) patients with no evidence of disease (DF), 135 advanced non-small cell lung cancer (NSCLC), 51 small cell lung cancer (SCLC). SI markers neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte (PLR), systemic inflammation index (SII), and systemic inflammation response index (SIRI) were determined from blood counts. Forty-seven plasma cytokines were measured using a multiplex bead-based assay. Progression-free survival (PFS) and overall survival (OS) were assessed using Kaplan-Meier and Cox Proportional Hazards models. NSCLC patients had significantly high levels of SI markers than SCLC and DF patients, while NLR, PLR and SII were also higher in SCLC than DF patients. SI optimized marker values to differentiate SI value were; 6.04 (NLR), 320 (PLR), 1615 (SII), and 7.3 (SIRI). Elevated levels NLR (p<0.001), PLR (p<0.001), and SII (p = 0.018) were associated with a worse PFS and OS in NSCLC, while none of the markers were associated with PFS in SCLC patients. NSCLC patients with a poor outcome displayed heterogeneous immune states relative to systemic inflammation and circulating IL-6 markers. These groups could be distinguished based on the cytokines IL-8, TNFα, and IL-27. We identified heterogeneity of immune states in SCLC and NSCLC patients and in NSCLC patients with the poorest prognosis. This heterogeneity could be exploited to improve outcomes for these patients.
    DOI:  https://doi.org/10.1371/journal.pone.0260988
  13. BMC Med Genomics. 2021 Dec 03. 14(1): 286
    Identification of the ferroptosis-related long non-coding RNAs signature to improve the prognosis prediction and immunotherapy response in patients with NSCLC.
    Meng Li, Yanpeng Zhang, Meng Fan, Hui Ren, Mingwei Chen, Puyu Shi.
      BACKGROUND: Non-small cell lung cancer (NSCLC) is the most prevalent type of lung carcinoma with an unfavorable prognosis. Ferroptosis is involved in the development of multiple cancers. Whereas, the prognostic value of ferroptosis-related lncRNAs in NSCLC remains uncertain.METHODS: Gene expression profiles and clinical information of NSCLC were retrieved from the TCGA database. Ferroptosis-related genes (FRGs) were explored in the FerrDb database and previous studies, ferroptosis-related lncRNAs (FRGs-lncRNAs) were identified by the correlation analysis and the LncTarD database. The differentially expressed FRGs-lncRNAs were screened and FRGs-lncRNAs associated with the prognosis were explored by univariate Cox regression analysis and Kaplan-Meier survival analysis. Then, an FRGs-lncRNAs signature was constructed and verified by the Lasso-penalized Cox analysis. Finally, the potential correlation between risk score, immune checkpoint genes, and chemotherapeutic sensitivity was further investigated.
    RESULTS: 129 lncRNAs with a potential regulatory relationship with 59 differentially expressed FRGs were found in NSCLC, of which 10 were related to the prognosis of NSCLC (P < 0.05). 9 prognostic-related FRGs-lncRNAs were used to construct the prognostic model and stratify NSCLC patients into high- and low-risk groups. A worse outcome was found in patients with high risk (P < 0.05). Moreover, a good predictive capacity of this signature in predicting NSCLC prognosis was confirmed. Additionally, 45 immune checkpoint genes and 4 chemotherapeutics drugs for NSCLC were identified to be correlated with the risk score.
    CONCLUSION: A novel FRGs-lncRNAs signature was successfully constructed, which may contribute to improving the management strategies of NSCLC.
    Keywords:  Ferroptosis; Long non-coding RNAs; Non-small cell lung cancer; Prognostic; Signature
    DOI:  https://doi.org/10.1186/s12920-021-01133-4
  14. Dis Markers. 2021 ;2021 4495489
    Derivation of a Novel CIHI in Patients with Lung Adenocarcinoma for Estimating Tumor Microenvironment and Clinical Prognosis.
    Liang Zhou, Guangyan Xu, Li Wang, Jianyong Zhang, Weimin Li.
      An interaction between hypoxia and immunity has been confirmed in tumor tissue. However, there is no combined biomarker for diagnosis on this basis. Therefore, we developed a scoring formula based on markers of hypoxia and immunity. Firstly, the hypoxia-immune formula of lung adenocarcinoma (LUAD) was derived using LASSO-Cox regression in three cohorts from public database, and the corresponding score was calculated for each patient. The formula is as follows: combined hypoxia and immune index (CIHI) = LDHA expression × 0.2252 + GAPDH expression × 0.0727 + ANGPTL4 expression × 0.0724 + VEGFC expression × 0.1911 + DKK1 expression × 0.1355 + ADM expression × 0.0588 + BTK expression × -0.1659. Meanwhile, patients were divided into groups according to high and low CIHI, and expression profiles of hypoxia markers and immune markers were analyzed in different groups. CIHI was used to confirm that patients with high CIHI represented a state of hypoxiahigh-immunitylow, which had worse overall survival. We also discussed the evaluation value in the immune microenvironment and clinical application of CIHI. In conclusion, this study developed and validated a hypoxia-immune formula that can guide hypoxia modifier treatment and immunotherapy in LUAD.
    DOI:  https://doi.org/10.1155/2021/4495489
  15. J Cancer Res Clin Oncol. 2021 Nov 28.
    Subtyping non-small cell lung cancer by histology-guided spatial metabolomics.
    Judith Martha Neumann, Hinrich Freitag, Jasmin Saskia Hartmann, Karsten Niehaus, Michail Galanis, Martin Griesshammer, Udo Kellner, Hanna Bednarz.
      PURPOSE: Most cancer-related deaths worldwide are associated with lung cancer. Subtyping of non-small cell lung cancer (NSCLC) into adenocarcinoma (AC) and squamous cell carcinoma (SqCC) is of importance, as therapy regimes differ. However, conventional staining and immunohistochemistry have their limitations. Therefore, a spatial metabolomics approach was aimed to detect differences between subtypes and to discriminate tumor and stroma regions in tissues.METHODS: Fresh-frozen NSCLC tissues (n = 35) were analyzed by matrix-assisted laser desorption/ionization-mass spectrometry imaging (MALDI-MSI) of small molecules (< m/z 1000). Measured samples were subsequently stained and histopathologically examined. A differentiation of subtypes and a discrimination of tumor and stroma regions was performed by receiver operating characteristic analysis and machine learning algorithms.
    RESULTS: Histology-guided spatial metabolomics revealed differences between AC and SqCC and between NSCLC tumor and tumor microenvironment. A diagnostic ability of 0.95 was achieved for the discrimination of AC and SqCC. Metabolomic contrast to the tumor microenvironment was revealed with an area under the curve of 0.96 due to differences in phospholipid profile. Furthermore, the detection of NSCLC with rarely arising mutations of the isocitrate dehydrogenase (IDH) gene was demonstrated through 45 times enhanced oncometabolite levels.
    CONCLUSION: MALDI-MSI of small molecules can contribute to NSCLC subtyping. Measurements can be performed intraoperatively on a single tissue section to support currently available approaches. Moreover, the technique can be beneficial in screening of IDH-mutants for the characterization of these seldom cases promoting the development of treatment strategies.
    Keywords:  Adenocarcinoma; Isocitrate dehydrogenase; Mass spectrometry imaging; Metabolomics; Non-small cell lung cancer; Squamous cell carcinoma
    DOI:  https://doi.org/10.1007/s00432-021-03834-w
  16. Front Oncol. 2021 ;11 706409
    Survival Outcomes of Local Compared With Systemic First Treatment of Non-Small Cell Lung Cancer Brain Metastases.
    Hong-Mei Liu, Chun-Liu Meng, Lu-Jun Zhao.
      Objective: This retrospective study evaluated the survival advantage of local treatment targeted to brain metastases, relative to systemic therapy, as the first option for brain metastases of non-small cell lung cancer (NSCLC).Methods: First reviewed were 291 cases of NSCLC brain metastases from two centers. All patients were at least 18 years old, with histologically confirmed NSCLC, and required and underwent both local (radiotherapy or brain surgery) and systemic treatment (chemotherapy and tyrosine kinase inhibitor [TKI] medication). Demographics, clinical characteristics, and treatment-related variables were collected.
    Results: The final population comprised 160 patients. Overall, the multivariate analysis suggested that the following were associated with better survival: >3 cycles of chemotherapy; stereotactic radiosurgery; and TKI medication (all, P = 0.000). Local treatment that began within 1 week of the diagnosis of brain metastases was associated with poorer survival (P = 0.006). Among the 111 patients with symptomatic brain metastases, the multivariate analysis indicated that better survival was associated with >3 cycles of chemotherapy (P = 0.000), radiation dose >40 Gy (P = 0.001), stereotactic radiosurgery (P = 0.000), and TKI medication (P = 0.000), while local treatment that began within 1 week after the diagnosis of brain metastases was associated with poorer survival (P = 0.015).
    Conclusions: For patients with NSCLC brain metastases, regardless of the presence of clinical symptoms associated with brain metastases, systemic treatment before local may be better for survival. Even when used to relieve clinical symptoms, local treatment should be within a setting of sufficient systemic treatment.
    Keywords:  brain metastases; initiation of local treatment; local treatment; non-small cell lung cancer; systemic treatment
    DOI:  https://doi.org/10.3389/fonc.2021.706409
  17. Cancer Treat Res Commun. 2021 Nov 10. pii: S2468-2942(21)00184-2. [Epub ahead of print]29 100488
    The impact of pathology grossing protocol measures to improve pathologic nodal staging in lung cancer.
    Robert Ta, Gracijela Bozovic, Jennifer L Wilson, Alexander A Bankier, Paul A VanderLaan.
      OBJECTIVE: Accurate assessment of lymph node (LN) status is essential for proper staging of resected lung cancer specimens. Here, we assessed pathology-centric interventions to increase the number of peribronchial LNs identified and evaluated in anatomic lung cancer resection specimens as part of a quality improvement initiative.MATERIALS AND METHODS: All non-small cell lung cancer (NSCLC) anatomic resection specimens from 2017 to 2020 were evaluated, comprising two years pre-intervention and one year post-intervention. We instituted 3 measures to increase peribronchial LN yield: 1) educational grossing sessions for pathology assistants and residents, 2) directions to submit additional peribronchial tissue if no LNs were identified grossly, and 3) a hard-stop prior to sign-out by the attending pathologist if no peribronchial LNs were identified.
    RESULTS: Of the total 227 resection specimens for NSCLC, 107/151 (70.9%) of specimens prior to the intervention had peribronchial LNs identified, whereas after the intervention significantly more (66/76, 86.8%, p < 0.01) specimens had peribronchial LNs identified. In addition, the mean number of peribronchial LNs identified significantly increased from 2.7 ± 3.3 pre-intervention to 4.3 ± 4.0 post-intervention (p < 0.001). Further analysis revealed a strong correlation between peribronchial LN metastases with both overall tumor size and invasive component size (for adenocarcinomas), correlation coefficient 0.974, p < 0.0001.
    CONCLUSION: Establishing focused grossing measures by pathology led to a significant increase in the number of peribronchial LNs identified and assessed during histopathologic evaluation of anatomic lung cancer resection specimens. Larger tumors are more likely to have occult peribronchial LN metastases, which may warrant more aggressive peribronchial LN assessment for larger tumors.
    Keywords:  Lymph node staging; Non-small cell lung cancer; Pathology grossing; Quality improvement
    DOI:  https://doi.org/10.1016/j.ctarc.2021.100488
  18. Sci Rep. 2021 Dec 02. 11(1): 23331
    The association between gut microbiome affecting concomitant medication and the effectiveness of immunotherapy in patients with stage IV NSCLC.
    M V Verschueren, C M Cramer- van der Welle, M Tonn, F M N H Schramel, B J M Peters, E M W van de Garde.
      Several observational studies suggested that gut microbiome-affecting-medication impairs the effectiveness of immunotherapy in patients with metastatic non-small-cell lung cancer (NSCLC). We postulated that if the effectiveness of immunotherapy is affected by drug-related changes of the microbiome, a stronger association between the use of co-medication and overall survival (OS) will be observed in patients treated with immunotherapy as compared to patients treated with chemotherapy. In a retrospective matched cohort study, immunotherapy patients were matched (1:1) to patients treated with chemotherapy in the pre immunotherapy era. The association between the use of antibiotics, opioids, proton pump inhibitors, metformin and other antidiabetics on OS was assessed with multivariable cox-regression analyses. Interaction tests were applied to investigate whether the association differs between patients treated with immuno- or chemotherapy. A total of 442 patients were studied. The use of antibiotics was associated with worse OS (adjusted Hazard Ratio (aHR) 1.39, p = 0.02) independent of the type of therapy (chemotherapy or immunotherapy). The use of opioids was also associated with worse OS (aHR 1.33, p = 0.01). The other drugs studied showed no association with OS. Interaction term testing showed no effect modification by immuno- or chemotherapy for the association of antibiotics and opioids with OS. The use of antibiotics and opioids is similarly associated with worse outcomes in both chemotherapy and immunotherapy treated NSCLC patients. This suggests that the association is likely to be a consequence of confounding rather than disturbing the composition of the microbiome.
    DOI:  https://doi.org/10.1038/s41598-021-02598-0
  19. Cell Death Dis. 2021 Dec 03. 12(12): 1127
    Amino acid deprivation induces AKT activation by inducing GCN2/ATF4/REDD1 axis.
    Hyeon-Ok Jin, Sung-Eun Hong, Ji-Young Kim, Se-Kyeong Jang, In-Chul Park.
      Amino acid availability is sensed by various signaling molecules, including general control nonderepressible 2 (GCN2) and mechanistic target of rapamycin complex 1 (mTORC1). However, it is unclear how these sensors are associated with cancer cell survival under low amino acid availability. In the present study, we investigated AKT activation in non-small cell lung cancer (NSCLC) cells deprived of each one of 20 amino acids. Among the 20 amino acids, deprivation of glutamine, arginine, methionine, and lysine induced AKT activation. AKT activation was induced by GCN2/ATF4/REDD1 axis-mediated mTORC2 activation under amino acid deprivation. In CRISPR-Cas9-mediated REDD1-knockout cells, AKT activation was not induced by amino acid deprivation, indicating that REDD1 plays a major role in AKT activation under amino acid deprivation. Knockout of REDD1 sensitized cells cultured under glutamine deprivation conditions to radiotherapy. Taken together, GCN2/ATF4/REDD1 axis induced by amino acid deprivation promotes cell survival signal, which might be a potential target for cancer therapy.
    DOI:  https://doi.org/10.1038/s41419-021-04417-w
  20. Carcinogenesis. 2021 Nov 19. pii: bgab104. [Epub ahead of print]
    Functional assessment of somatic STK11 variants identified in primary human non-small cell lung cancers.
    Liam L Donnelly, Tyler C Hogan, Sean M Lenahan, Gopika Nandagopal, Jenna G Eaton, Meagan A Lebeau, Cai L McCann, Hailey M Sarausky, Kenneth J Hampel, Jordan D Armstrong, Margaret P Cameron, Nikoletta Sidiropoulos, Paula Deming, David J Seward.
      Serine/Threonine Kinase 11 (STK11) encodes an important tumor suppressor that is frequently mutated in lung adenocarcinoma. Clinical studies have shown that mutations in STK11 resulting in loss of function correlate with resistance to anti-PD-1 monoclonal antibody therapy in KRAS-driven non-small cell lung cancer (NSCLC), but the molecular mechanisms responsible remain unclear. Despite this uncertainty, STK11 functional status is emerging as a reliable biomarker for predicting non-response to anti-PD-1 therapy in NSCLC patients. The clinical utility of this biomarker ultimately depends upon accurate classification of STK11 variants. For nonsense variants occurring early in the STK11 coding region, this assessment is straightforward. However, rigorously demonstrating the functional impact of missense variants remains an unmet challenge. Here we present data characterizing four STK11 splice-site variants by analyzing tumor mRNA, and 28 STK11 missense variants using an in vitro kinase assay combined with a cell-based p53-dependent luciferase reporter assay. The variants we report were identified in primary human NSCLC biopsies in collaboration with the University of Vermont Genomic Medicine group. Additionally, we compare our experimental results with data from 22 in silico predictive algorithms. Our work highlights the power, utility and necessity of functional variant assessment and will aid STK11 variant curation, provide a platform to assess novel STK11 variants and help guide anti-PD-1 therapy utilization in KRAS-driven NSCLCs.
    DOI:  https://doi.org/10.1093/carcin/bgab104
  21. Leuk Lymphoma. 2021 Nov 29. 1-8
    The Prognostic Nutritional Index (PNI) is an independent predictor of overall survival in older patients with follicular lymphoma.
    Pablo Mozas, Andrea Rivero, Alfredo Rivas-Delgado, Ferran Nadeu, Eva Giné, Julio Delgado, Neus Villamor, Elías Campo, Patricia Pérez-Galán, Laura Magnano, Armando López-Guillermo.
      The Prognostic Nutritional Index (PNI), a parameter combining serum albumin concentration and absolute lymphocyte count, is considered a measure of the nutritional and inflammatory status and the host's anti-tumor response. We analyzed the clinical characteristics and outcomes according to the PNI of 351 grades 1-3 A FL patients. Forty-one patients (12%) had a PNI ≤45, who were older and showed adverse baseline features. A low PNI was associated with a shorter PFS (only for patients >60 years), and OS (for all patients, 10-year OS, 52% versus 74%, p = 0.0001). The prognostic impact of the PNI on OS was confirmed in a multivariate model for patients >60 years (HR = 3, p = 0.006). In conclusion, the PNI is a readily accessible piece of information that can identify a small subset of FL patients with shorter survival, and it could be an aid to improve the nutritional status of patients prior to treatment initiation.
    Keywords:  Follicular lymphoma; Prognostic Nutritional Index; chemoimmunotherapy; response; survival
    DOI:  https://doi.org/10.1080/10428194.2021.2010064
  22. Front Immunol. 2021 ;12 697298
    Immune Checkpoint Inhibitors With or Without Bone-Targeted Therapy in NSCLC Patients With Bone Metastases and Prognostic Significance of Neutrophil-to-Lymphocyte Ratio.
    Alberto Bongiovanni, Flavia Foca, Jessica Menis, Stefania Luigia Stucci, Fabrizio Artioli, Valentina Guadalupi, Maria Rosachiara Forcignanò, Manuela Fantini, Federica Recine, Laura Mercatali, Chiara Spadazzi, Marco Angelo Burgio, Valentina Fausti, Anna Miserocchi, Toni Ibrahim.
      Introduction: Bone metastases (BMs) are a negative prognostic factor in patients with non-small cell lung cancer (NSCLC). Although immune-checkpoint inhibitors (ICIs) have dramatically changed the therapeutic landscape of NSCLC, little information is available on BMs from NSCLC treated with ICIs alone or in association with bone-targeted therapy (BTT) such as zoledronate or denosumab.Methods: From 2014 to 2020, 111 of the 142 patients with BMs secondary to NSCLC extrapolated from the prospective multicenter Italian BM Database were eligible for analysis. Information on blood count, comorbidities, and toxicity was retrospectively collected. The neutrophil-to-lymphocyte ratio (NLR) pre- and post-treatment was calculated. Survival was analyzed using the Kaplan-Meier method, with statistical significance of survival differences assessed using the log-rank test.
    Results: Median age was 66 (range, 42-84) years. Performance status (PS) Eastern Cooperative Oncology Group (ECOG) was 0-1 in 79/111 patients. The majority of patients (89.2%) had adenocarcinoma histology. At a median follow-up of 47.4 months, median progression-free (mPFS) and overall survival (mOS) was 4.9 (95%CI, 2.8-10.0) and 11.9 (95%CI, 8.2-14.4) months, respectively. Forty-six (43.4%) patients with BM NSCLC underwent first- or further-line therapy with ICIs: 28 (60.8%) received nivolumab, 9 (19.6%) pembrolizumab, and 9 (19.6%) atezolizumab. Of the 46 patients treated with ICIs, 30 (65.2%) underwent BTT: 24 (80.0%) with zoledronate and 6 (20.0%) with denosumab. The ICI-alone group had an mOS of 15.8 months [95%CI, 8.2-not evaluable (NE)] vs. 21.8 months (95%CI, 14.5-not evaluable) for the ICI plus BTT group and 7.5 (95%CI, 6.1-10.9) months for the group receiving other treatments (p < 0.001). NLR ≤5 had a positive impact on OS.
    Conclusion: BTT appears to have a synergistic effect when used in combination with ICIs, improving patient survival.
    Keywords:  NSCLC; bone metastases; denosumab; immune checkpoint inhibitors; lung cancer; zoledronate
    DOI:  https://doi.org/10.3389/fimmu.2021.697298
  23. Pharmacol Rep. 2021 Nov 30.
    Decreased efficacy of the ketamine and scopolamine-induced sustained antidepressant-like effects in rats receiving metformin.
    Han-Wen Chuang, I-Hua Wei, Chun-Te Li, Chih-Chia Huang.
      BACKGROUND: Metformin is the most widely used drug for treating type 2 diabetes mellitus (DM), which frequently co-occurs with depressive disorders. Thus, patients with depression are likely to receive metformin. Metformin activates AMP-activated kinase (AMPK), which inhibits mechanistic target of rapamycin complex 1 (mTORC1) signaling. mTORC1 activation is essential for the antidepressant effects of ketamine and scopolamine. Thus, we hypothesized that metformin may attenuate ketamine- or scopolamine-induced antidepressant efficacies by blocking their mTORC1 activation.METHODS: We assessed the acute and sustained antidepressant-like actions of ketamine and scopolamine in male Sprague-Dawley rats subjected to the forced swim test with or without metformin pretreatment. The expressions of AMPK, mTORC1, and brain-derived neurotrophic factor (BDNF) in their prefrontal cortex were assessed.
    RESULTS: Metformin (50 mg/kg) attenuated the sustained, but not acute, antidepressant-like effects of ketamine (10 mg/kg) and scopolamine (25 μg/kg). Although metformin reduced mTORC1 downstream activated P70S6K, it did not significantly alter mTORser2448 activation and even increased BDNF expression. Notably, ketamine, scopolamine, and metformin all exerted significant antidepressant-like actions, as evidenced by increased AMPK phosphorylation and BDNF expression.
    CONCLUSIONS: Metformin-induced attenuation of sustained antidepressant-like effects are not directly dependent on AMPK-deactivated mTORC1. Our results indicate the complexity of interactions between AMPK, BDNF, and mTORC1. Further research, including mechanistic studies, is warranted to comprehensively evaluate the application of metformin in patients receiving mTORC1-based antidepressants.
    Keywords:  AMPK; BDNF; Ketamine; Metformin; Scopolamine; mTOR
    DOI:  https://doi.org/10.1007/s43440-021-00342-z
  24. Clin Lung Cancer. 2021 Oct 25. pii: S1525-7304(21)00274-6. [Epub ahead of print]
    Overall Survival and Safety With Pemetrexed/Platinum ± Anti-VEGF Followed by Pemetrexed ± Anti-VEGF Maintenance in Advanced Nonsquamous Non-Small-Cell Lung Cancer: A Pooled Analysis of 4 Randomized Studies.
    Edward B Garon, Patrick Peterson, Maria Teresa Rizzo, Jong Seok Kim.
      BACKGROUND: Before immune checkpoint blockade therapy, chemotherapy with pemetrexed maintenance was the standard of care for patients with advanced nonsquamous non-small-cell lung cancer (NSQ-NSCLC) and remains such where immunotherapy is not applicable. This pooled analysis aimed to characterize overall survival (OS) and safety of pemetrexed ± anti-VEGF maintenance, by treatment duration.PATIENTS AND METHODS: Data from 4 randomized clinical trials (PARAMOUNT, PRONOUNCE, PointBreak, JVBL) of patients with NSQ-NSCLC receiving pemetrexed ± anti-VEGF maintenance therapy were pooled as 2 groups (Group A: pemetrexed-only maintenance, n = 486; and Group B: pemetrexed + anti-VEGF maintenance, n = 329). OS and treatment-emergent adverse events (TEAEs) were analyzed in both groups by treatment duration.
    RESULTS: Baseline characteristics were well balanced between both groups. Median OS did not significantly differ between Group A (16.1 months) and Group B (18.4 months; hazard ratio: 1.17, P= .1417). A correlation between median OS and treatment duration was numerically stronger in Group A (r = 0.72) versus B (r = 0.62). Across treatment groups, TEAEs were largely grade 1 to 2 and, with few exceptions, did not increase with increased treatment duration.
    CONCLUSION: There was no significant OS difference between pemetrexed-only and pemetrexed ± anti-VEGF maintenance in patients with NSQ-NSCLC. Patients receiving pemetrexed + anti-VEGF experienced a slightly less favorable safety profile with more reported TEAEs compared to pemetrexed monotherapy. Pemetrexed ± anti-VEGF maintenance therapy may be considered in NSQ-NSCLC, based on an individualized patient approach, particularly where immunotherapy is not clinically indicated.
    Keywords:  Clinical trial outcomes; Maintenance therapy; NSCLC; Safety; Survival
    DOI:  https://doi.org/10.1016/j.cllc.2021.10.010
  25. J Lipid Res. 2021 Nov 24. pii: S0022-2275(21)00137-1. [Epub ahead of print] 100154
    1-deoxysphinganine initiates adaptive responses to serine and glycine starvation in cancer cells via proteolysis of sphingosine kinase.
    Jean-Philip Truman, Christian F Ruiz, Emily Montal, Monica Garcia-Barros, Izolda Mileva, Ashley J Snider, Yusuf A Hannun, Lina M Obeid, Cungui Mao.
      Cancer cells can become dependent on exogenous serine, depletion of which results in slower growth and activation of a number of adaptive metabolic changes. We previously demonstrated that serine and glycine (SG) deprivation causes loss of sphingosine kinase 1 (SK1) in cancer cells, thereby increasing levels of its lipid substrate, sphingosine (Sph), which mediates several adaptive biological responses. However, the signaling molecules that regulate levels of SK1 and Sph in response to SG deprivation have yet to be defined. Here, we identify 1-deoxysphinganine (dSA), a non-canonical sphingoid base generated in the absence of serine from the alternative condensation of alanine and palmitoyl CoA by serine palmitoyl transferase (SPT), as a proximal mediator of SG deprivation in SK1 loss and Sph level elevation in SG deprivation in cancer cells. SG starvation markedly increased dSA levels in vitro and in vivo, and in turn induced SK1 degradation through a SPT-dependent mechanism, resulting in an increase in SPH levels. Addition of exogenous dSA caused a moderate increase in intracellular reactive oxygen species (ROS), which in turn decreased pyruvate kinase PKM2 activity while increasing phosphoglycerate dehydrogenase (PHGDH) levels, and thereby promoted serine synthesis. We further showed that increased dSA induces the adaptive cellular and metabolic functions in the response of cells to decreased availability of serine likely by increasing Sph levels. Thus, we conclude that dSA functions as an initial sensor of serine loss, SK1 functions as its direct target, and Sph functions as a downstream effector of cellular and metabolic adaptations. These studies define a previously unrecognized 'physiological' non-toxic function for dSA.
    Keywords:  Sphingosine kinase; hereditary sensory and autonomic neuropathy (HSAN); mass spectrometry; phosphoglycerate dehydrogenase (PHGDH); pyruvate kinase (PKM2); reactive oxygen species (ROS); serine biosynthesis; serine palmitoyl transferase (SPT); sphingosine; ubiquitination
    DOI:  https://doi.org/10.1016/j.jlr.2021.100154
  26. Cancer Cell Int. 2021 Nov 27. 21(1): 627
    Downregulation of miR-29c promotes muscle wasting by modulating the activity of leukemia inhibitory factor in lung cancer cachexia.
    Kairu Xie, Hairong Xiong, Wen Xiao, Zhiyong Xiong, Wenjun Hu, Jiaxin Ye, Ning Xu, Jian Shi, Changfei Yuan, Zhixian Chen, Daojia Miao, Xiaoping Zhang, Hongmei Yang.
      BACKGROUND: Cancer cachexia is a wasting disorder characterized by significant weight loss, and is attributed to skeletal muscle weakness. In the process of cancer development, microRNAs act as oncogenes or tumor suppressors. Moreover, they are implicated in muscle development and wasting. This study sought to explore the mechanisms and correlation between miR-29c and muscle wasting in lung cancer cachexia.METHODS: Data for expression analysis were retrieved from the Cancer Genome Atlas (TCGA) database. qRT-PCR analyses were performed to explore the expression levels of miR-29c and Leukemia Inhibitory Factor (LIF). Lewis lung carcinoma (LLC) cell line was used to establish a cachexia model to explore the functions of miR-29c and LIF in lung cancer cachexia. Furthermore, in vitro (in C2C12 myotubes) and in vivo (in LLC tumor-bearing mice) experiments were performed to explore the mechanisms of miR-29c and LIF in lung cachexia.
    RESULTS: Analysis of the lung cancer cachexia model showed that miR-29c was down-regulated, and its expression was negatively correlated with muscle catabolic activity. Overexpression of miR-29c mitigated the cachectic phenotype. Mechanistic studies showed that LIF was a direct target gene of miR-29c, and LIF was upregulated in vitro and in vivo. Analysis showed that LIF promoted muscle wasting through the JAK/STAT and MAP-kinase pathways.
    CONCLUSIONS: The findings indicated that miR-29c was negatively correlated with the cachectic phenotype, and the miR-29c-LIF axis is a potential therapeutic target for cancer cachexia.
    Keywords:  Cachexia; Leukemia inhibitory factor; Muscular atrophy; miR-29c
    DOI:  https://doi.org/10.1186/s12935-021-02332-w
  27. Bioengineered. 2021 Dec 02.
    Testis developmental related gene 1 promotes non-small-cell lung cancer through the microRNA-214-5p/Krüppel-like factor 5 axis.
    Xudong Lu, Nian Zhao, Guangjun Duan, Zhiyong Deng, Yimin Lu.
      FLOW SHEET ABSTRACT: [Formula: see text].
    Keywords:  KLF5; NSCLC; TDRG1; miR-214-5p
    DOI:  https://doi.org/10.1080/21655979.2021.2012406
  28. Brief Funct Genomics. 2021 Nov 25. pii: elab043. [Epub ahead of print]
    Seven autophagy-related lncRNAs are associated with the tumor immune microenvironment in predicting survival risk of nonsmall cell lung cancer.
    Huihui Jiang, Aiqun Xu, Min Li, Rui Han, Enze Wang, Dong Wu, Guanghe Fei, Sijing Zhou, Ran Wang.
      BACKGROUND: Nonsmall cell lung cancer (NSCLC) ranks first among global cancer-related deaths. Despite the emergence of various immunological and targeted therapies, immune tolerance remains a barrier to treatment.METHODS: It has been found that this obstacle can be overcome by targeting autophagy-related genes (ATGs). ATGs were screened by coexpression analysis and the genes related to the prognosis of lung cancer were screened using Kaplan-Meier (K-M) survival analysis, univariate Cox regression and multivariate Cox regression. The prognostic risk model of ATGs was constructed and verified using K-M survival analysis and receiver operating characteristic (ROC) curve analysis.
    RESULTS: The prognostic risk model of ATGs was constructed. Gene set enrichment analysis (GSEA) showed that the function and pathway of ATG enrichment were closely related to immune cell function. CIBERSORT, LM22 matrix and Pearson correlation analysis showed that risk signals were significantly correlated with immune cell infiltration and immune checkpoint genes.
    CONCLUSIONS: We identified and independently verified the ATG (AL691432.2, MMP2-AS1, AC124067.2, CRNDE, ABALON, AL161431.1, NKILA) in NSCLC patients and found that immune regulation in the tumor microenvironment is closely related to this gene.
    Keywords:  LncRNA; autophagy; immune microenvironment; nonsmall cell lung cancer
    DOI:  https://doi.org/10.1093/bfgp/elab043
  29. World J Surg Oncol. 2021 Nov 29. 19(1): 335
    Circular RNA circVAPA contributes to non-small-cell lung cancer progression via miR-342-3p-dependent regulation of ZEB2.
    Xiaoyang Liu, Yang Cheng, Yan Wang, Yinhong Zhang.
      BACKGROUND: Accumulating evidence demonstrated that circular RNAs (circRNAs) play pivotal regulatory roles in the pathology of cancers. Disclosing the roles and molecular mechanisms of circRNAs in tumorigenesis and development is essential to identify novel diagnostic and therapeutic targets. In this study, we explored the role of circVAPA in non-small-cell lung cancer (NSCLC) progression and its associated mechanism.METHODS: The expression level of RNA was analyzed by real-time quantitative polymerase chain reaction (RT-qPCR). Cell proliferation was assessed by MTT assay and colony-forming assay. Cell apoptosis was analyzed by flow cytometry. Cell migration and invasion were assessed by transwell assays. Dual-luciferase reporter, RNA pull-down, and RNA immunoprecipitation (RIP) assays were used to test the intermolecular interactions. The role of circVAPA was assessed in vivo. And xenograft tumor tissues were analyzed by immunohistochemistry (IHC) staining.
    RESULTS: CircVAPA expression was upregulated in NSCLC tissues and cell lines, and a high level of circVAPA was associated with a poor prognosis of NSCLC patients. CircVAPA silencing suppressed the proliferation, migration, and invasion and induced the apoptosis of NSCLC cells. CircVAPA served as a molecular sponge for microRNA-342-3p (miR-342-3p). miR-342-3p interference largely reversed circVAPA knockdown-mediated anti-tumor effects in NSCLC cells. Zinc finger E-box-binding homeobox 2 (ZEB2) was a target of miR-342-3p, and miR-342-3p overexpression suppressed the malignant behaviors of NSCLC cells largely by downregulating ZEB2. CircVAPA silence repressed xenograft tumor growth in vivo, and IHC assay confirmed that circVAPA silence restrained the proliferation and metastasis but induced the apoptosis of NSCLC cells in vivo.
    CONCLUSION: CircVAPA contributes to the progression of NSCLC by binding to miR-342-3p to upregulate ZEB2. CircVAPA/miR-342-3p/ZEB2 axis might be a novel potential target for NSCLC treatment.
    Keywords:  NSCLC; ZEB2; circVAPA; miR-342-3p
    DOI:  https://doi.org/10.1186/s12957-021-02447-4
  30. Natl Sci Rev. 2021 Oct;8(10): nwab014
    Targeting HSPA1A in ARID2-deficient lung adenocarcinoma.
    Xue Wang, Yuetong Wang, Zhaoyuan Fang, Hua Wang, Jian Zhang, Longfu Zhang, Hsinyi Huang, Zhonglin Jiang, Yujuan Jin, Xiangkun Han, Shenda Hou, Bin Zhou, Feilong Meng, Luonan Chen, Kwok-Kin Wong, Jinfeng Liu, Zhiqi Zhang, Xin Zhang, Haiquan Chen, Yihua Sun, Liang Hu, Hongbin Ji.
      Somatic mutations of the chromatin remodeling gene ARID2 are observed in ∼7% of human lung adenocarcinomas (LUADs). However, the role of ARID2 in the pathogenesis of LUADs remains largely unknown. Here we find that ARID2 expression is decreased during the malignant progression of both human and mice LUADs. Using two KrasG12D -based genetically engineered murine models, we demonstrate that ARID2 knockout significantly promotes lung cancer malignant progression and shortens overall survival. Consistently, ARID2 knockdown significantly promotes cell proliferation in human and mice lung cancer cells. Through integrative analyses of ChIP-Seq and RNA-Seq data, we find that Hspa1a is up-regulated by Arid2 loss. Knockdown of Hspa1a specifically inhibits malignant progression of Arid2-deficient but not Arid2-wt lung cancers in both cell lines as well as animal models. Treatment with an HSPA1A inhibitor could significantly inhibit the malignant progression of lung cancer with ARID2 deficiency. Together, our findings establish ARID2 as an important tumor suppressor in LUADs with novel mechanistic insights, and further identify HSPA1A as a potential therapeutic target in ARID2-deficient LUADs.
    Keywords:  ARID2; HSPA1A; lung adenocarcinoma; tumor suppressor
    DOI:  https://doi.org/10.1093/nsr/nwab014
  31. J Vet Sci. 2021 Nov;22(6): e92
    The effects of naringenin and naringin on the glucose uptake and AMPK phosphorylation in high glucose treated HepG2 cells.
    Lakshi A Dayarathne, Sachithra S Ranaweera, Premkumar Natraj, Priyanka Rajan, Young Jae Lee, Chang-Hoon Han.
      BACKGROUND: Naringin and its aglycone naringenin are citrus-derived flavonoids with several pharmacological effects. On the other hand, the mechanism for the anti-diabetic effects of naringenin and naringin are controversial and remain to be clarified further.OBJECTIVE: This study examined the relationship between glucose uptake and AMP-activated protein kinase (AMPK) phosphorylation by naringenin and naringin in high glucose-treated HepG2 cells.
    METHODS: Glucose uptake was measured using the 2-NBDG fluorescent D-glucose analog. The phosphorylation levels of AMPK and GSK3β (Glycogen synthase kinase 3 beta) were observed by Western blotting. Molecular docking analysis was performed to evaluate the binding affinity of naringenin and naringin to the γ-subunit of AMPK.
    RESULTS: The treatment with naringenin and naringin stimulated glucose uptake regardless of insulin stimulation in high glucose-treated HepG2 cells. Both flavonoids increased glucose uptake by promoting the phosphorylation of AMPK at Thr172 and increased the phosphorylation of GSK3β. Molecular docking analysis showed that both naringenin and naringin bind to the γ-subunit of AMPK with high binding affinities. In particular, naringin showed higher binding affinity than the true modulator, AMP with all three CBS domains (CBS1, 3, and 4) in the γ-subunit of AMPK. Therefore, both naringenin and naringin could be positive modulators of AMPK activation, which enhance glucose uptake regardless of insulin stimulation in high glucose-treated HepG2 cells.
    CONCLUSIONS: The increased phosphorylation of AMPK at Thr172 by naringenin and naringin might enhance glucose uptake regardless of insulin stimulation in high glucose treated HepG2 cells.
    Keywords:  AMPK phosphorylation; Naringenin; glucose uptake; molecular docking; naringin
    DOI:  https://doi.org/10.4142/jvs.2021.22.e92
  32. ESMO Open. 2021 Nov 24. pii: S2059-7029(21)00281-7. [Epub ahead of print]6(6): 100319
    Beyond epidermal growth factor receptor: MET amplification as a general resistance driver to targeted therapy in oncogene-driven non-small-cell lung cancer.
    N Coleman, L Hong, J Zhang, J Heymach, D Hong, X Le.
      The rapidly changing treatment paradigm for patients with metastatic oncogene-driven lung cancer continues to evolve, and consequently our understanding of the landscape of resistance must also advance. MET amplification is an established and frequent driver of resistance in EGFR-mutant non-small-cell lung cancer (NSCLC). Recently, the combination of MET proto-oncogene (MET) and epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) has shown promise in overcoming this molecularly defined resistance in clinical trials, and this combination strategy is being pursued in ongoing trials. Emerging data also demonstrate MET amplification as a resistance driver to TKI-treated ALK-, RET-, and ROS-1-fusion NSCLC, consistently at the range of 15%, while the resistance profiling data are maturing for other molecular targets. In this review, we discuss MET amplification as a driver of acquired resistance in well-defined molecular subsets of NSCLC, explore the biology behind this mechanism of resistance, and summarize the recently published clinical data, including the proposed combination strategies in the clinic achieving success in overcoming acquired MET amplification-dependent resistance.
    Keywords:  EGFR; MET; NSCLC; amplification; resistance; targeted therapy
    DOI:  https://doi.org/10.1016/j.esmoop.2021.100319
  33. BMC Res Notes. 2021 Nov 27. 14(1): 437
    Correlation of serum electrolytes with serial miRNA in advanced stage non-small cell lung cancer (NSCLC) in Indonesia.
    Arif Riswahyudi Hanafi, Achmad Mulawarman Jayusman, Priscillia Imelda, Serafim Alfasunu, Ahmad Hamim Sadewa, Dibyo Pramono, Didik Setyo Heriyanto, Sofia Mubarika Haryana, Siti Boedina Kresno.
      OBJECTIVE: This study aims to evaluate the correlation between electrolytes and serial miRNAs from our previous study. We want to prove that there is the molecular basis that underlying electrolytes disturbances as the predictive indicator to the outcome in NSCLC patients.RESULTS: There were positive correlation between potassium level with miR-34 (p  = 0.008, r  = 0.366), miR-148 (p  = 0.004, r  = 0.394) and miR-155 (p  = 0.031, r  = 0.300).
    Keywords:  Advanced stage NSCLC; Potassium; miRNA
    DOI:  https://doi.org/10.1186/s13104-021-05852-w
  34. J Oncol. 2021 ;2021 2721466
    Enhancing the Therapeutic Efficacy of KRASG12C Inhibitors in Lung Adenocarcinoma Cell Models by Cotargeting the MAPK Pathway or HSP90.
    Ying Liu, Lei Wu, Hong Lu, En Wu, Jun Ni, Xiaorong Zhou.
      Background: KRASG12C inhibitors have shown promising efficacy in early clinical trials, but drug resistance compromises their long-term benefits. Therefore, it is critical to understand the mechanisms of drug resistance and to design appropriate combinatory treatments to improve efficacy.Methods: To understand the comprehensive mechanisms of drug resistance, we treated lung cancer cells with KRASG12C inhibitors for different periods and performed transcriptional profiling and signaling analysis to identify critical factors and pathways that drive drug tolerance and resistance. We also evaluated several drug combinations in vitro and in vivo to identify potentially effective therapeutics.
    Results: We found that the feedback activation of multiple receptor tyrosine kinases (RTKs) may have cooperatively induced intrinsic and adaptive resistance to KRASG12C inhibitors. Notably, continuous KRAS inhibition induced a multidrug-resistant phenotype, implying that upfront combinatory treatment might be required to treat this group of patients. We also demonstrated that concurrently targeting multiple nodes in the RTK/RAS/RAF/MEK/ERK axis improved the efficacy of KRASG12C inhibitors, mainly by suppressing the reactivation of the mitogen-activated protein kinase (MAPK) pathway. Moreover, the combined use of HSP90 and KRASG12C inhibitors effectively induced tumor regression in lung adenocarcinoma models in vitro and in vivo.
    Conclusion: Together, our findings revealed mechanisms underlying KRASG12C inhibitors resistance and provided novel candidate combinatory strategies to improve their anticancer activity.
    DOI:  https://doi.org/10.1155/2021/2721466
  35. Int J Gen Med. 2021 ;14 8541-8555
    LncRNA ADAMTS9-AS2 is a Prognostic Biomarker and Correlated with Immune Infiltrates in Lung Adenocarcinoma.
    Zhichao Lin, Wenhai Huang, Yongsheng Yi, Dongbing Li, Zehua Xie, Zumei Li, Min Ye.
      Background: The role of long noncoding RNA (LncRNA) ADAMTS9 antisense RNA 2 (ADAMTS9-AS2) is unclear in lung adenocarcinoma (LUAD). The aim of this study was to explore the relationship between ADAMTS9-AS2 and LUAD, based on The Cancer Genome Atlas (TCGA) database and bioinformatics analysis.Methods: Various statistical methods, Kaplan-Meier method, Cox regression analysis, GSEA, and immune infiltration analysis were used to evaluate the relationship between clinical features and ADAMTS9-AS2 expression, prognostic factors, and the significant involvement of ADAMTS9-AS2 in function.
    Results: In LUAD patients, low expression of ADAMTS9-AS2 was associated with N stage (P=0.011), gender (P=0.002), number of packs smoked (P=0.024) and smoker (P<0.001). Low ADAMTS9-AS2 expression predicted a poorer overall survival (OS) (HR: 0.68; 95% CI: 0.51-0.91; P=0.01). And ADAMTS9-AS2 expression (HR: 0.626; 95% CI: 0.397-0.986; P=0.043) was independently correlated with OS in LUAD patients. Unwinding of DNA, extrinsic pathway, polo-like kinase-mediated events, cori cycle, MCM pathway, proteasome pathway, lagging strand synthesis and PCNA-dependent long patch base excision repair were differentially enriched in ADAMTS9-AS2 high expression phenotype. ADAMTS9-AS2 expression was correlated with certain immune infiltrating cells.
    Conclusion: In LUAD patients, ADAMTS9-AS2 expression was significantly associated with poor survival and immune infiltration. ADAMTS9-AS2 may be a promising biomarker of prognosis and response to immunotherapy for LUAD.
    Keywords:  ADAMTS9-AS2 antisense RNA 2; biomarker; immune infiltration; lung adenocarcinoma; prognosis
    DOI:  https://doi.org/10.2147/IJGM.S340683
  36. J Cell Mol Med. 2021 Dec 01.
    VLX1570 regulates the proliferation and apoptosis of human lung cancer cells through modulating ER stress and the AKT pathway.
    Juan Wang, Tongde Du, Ya Lu, Yan Lv, Yuxin Du, Jianzhong Wu, Rong Ma, Chenxin Xu, Jifeng Feng.
      The ubiquitin-proteasome system (UPS) possesses unique functions in tumorigenesis and has great potential for targeting tumours. The effectiveness of inhibitors targeting UPS in solid tumours remains to be studied. We screened a library of inhibitors targeting the ubiquitination system and found the highly potent, low-concentration inhibitor molecule VLX1570 that specifically killed lung cancer cells. VLX1570 is an inhibitor of deubiquitinating enzyme activity and has also shown potential for preclinical cancer treatment. We found that VLX1570 significantly inhibited lung cancer cells proliferation and colony formation. VLX1570 induced a G2/M cell cycle arrest by downregulating CDK1 and CyclinB1. Moreover, VLX1570 significantly promoted the mitochondrial-associated apoptosis. Mechanistically speaking, VLX1570 activated the PERK/IRE1/ATF6 pathway and induced ER stress in lung cancer cell lines. The inhibition of ER stress by tauroursodeoxycholic acid sodium or 4-phenylbutyric acid enhanced VLX1570-induced apoptosis. In addition, VLX1570 treatment led to the inactivation of Akt signalling and inhibited the proliferation of lung cancer cells by downregulating the Akt pathway. Moreover, combined treatment with VLX1570 and Afatinib or Gefitinib induced synergistic anti-lung cancer activity. Our present study demonstrated a novel therapy using VLX1570, which inhibited the proliferation and increased apoptosis in human lung cancer.
    Keywords:  AKT pathway; ER stress; VLX1570; apoptosis; lung cancer; proliferation
    DOI:  https://doi.org/10.1111/jcmm.17053
  37. J Cancer Res Ther. 2021 Nov;17(5): 1157-1164
    Effects of CT images at different reconstruction energies on radiotherapy planning of patients diagnosed with nonsmall cell lung cancer.
    Tian Xiufang, Liu Kun, Wang Jing, Li Cuihua, Zhang Jiandong, Yong Hou.
      Objective and Aims: We conducted this study to explore the influence of spectral computer tomography (CT) images at different reconstruction energies on the radiotherapy plan of patients with nonsmall-cell lung cancer (NSCLC).Subject and Methods: Here, 38 NSCLC patients were selected to undergo energy spectral scanning. All energy spectral images obtained were then transferred to the Discover™ CT postprocessing workstation to generate 40k eV, 60 keV, 80keV, 100keV, 120keV, and different 140keV single-energy images. Subsequently, the images were imported to the Eclipse planning system, after which an oncologist contoured the target area and organs at risk (OARs) on these single-energy images described above. Furthermore, a physicist then designed radiotherapy plans to conduct statistical analysis on the tissue CT value and target volume of each single-energy image, to compare the dosimetry of different plans about the OARs and the target area.
    Results: The CT values of gross tumor volumes (GTV), heart, lung, and spinal cord samples subjected to different energy CT images were statistically different (P < 0.05). Among them, the CT value of each tissue obtained in the 40 keV group was the largest and decreased with the increase in energy. As shown, no statistically significant differences were observed in the homogeneity index and conformity index, including the maximum, minimum, and average doses of GTV delineated on the CT images of different energies (P > 0.05), as well as the OARs.
    Conclusions: When CT images of different energies obtained from the energy spectral CT scans were used in the design of radiotherapy planning, no significant differences were observed in the target area outlines and in the doses caused by energy factors. However, the differences in tissue CT values had statistical significance.
    Keywords:  Dosimetry; energy spectrum computed tomography; nonsmall cell lung cancer; radiotherapy
    DOI:  https://doi.org/10.4103/jcrt.jcrt_1030_21
  38. Nutr Rev. 2021 Nov 28. pii: nuab090. [Epub ahead of print]
    Methods of nutritional assessment and functional capacity in the identification of unfavorable clinical outcomes in hospitalized patients with cancer: a systematic review.
    Mariana S Crestani, Thaiciane Grassi, Thais Steemburgo.
      CONTEXT: Malnutrition has a negative impact on patients with cancer. Identifying risk, nutritional status, and functional capacity can contribute to adequate and early nutritional therapy, which can reduce unfavorable clinical outcomes.OBJECTIVE: To evaluate and summarize the main instruments of nutritional assessment and functional capacity and associate their results with clinical outcomes in hospitalized patients with cancer.
    DATA SOURCES: A systematic search was performed in the PubMed/MEDLINE, Embase, SciELO, and LILACS databases. Studies in which researchers evaluated and compared screening, nutritional assessment, and functional capacity instruments and their associations with clinical outcomes were included.
    DATA EXTRACTION: The data were extracted by 2 independent reviewers.
    RESULTS: A total of 29 studies met the inclusion criteria (n = 20 441 individuals). The Nutritional Risk Screening-2002 (NRS-2002) and Patient-Generated Subjective Global Assessment (PG-SGA) were the most common tools used for nutritional assessment. High nutritional risk according to the NRS-2202 and worse nutritional status according to the PG-SGA and Subjective Global Assessment were positively associated with a longer hospital stay and mortality. Low functional capacity, according to handgrip strength, was associated with longer hospital stay and nutrition impact symptoms.
    CONCLUSIONS: Tools such as the NRS-2002, PG-SGA, Subjective Global Assessment, and handgrip strength assessment are efficacious for assessing unfavorable clinical outcomes in hospitalized patients with cancer.
    Keywords:  cancer; functional capacity; nutritional assessment; nutritional screening; unfavorable clinical outcomes
    DOI:  https://doi.org/10.1093/nutrit/nuab090
  39. Thorac Cancer. 2021 Dec 02.
    Inhibitory effects of icotinib combined with antiangiogenic drugs in human non-small cell lung cancer xenograft models are better than single target drugs.
    Peng Jiang, Yan Zhang, Jiadong Cui, Xiuxiu Wang, Yu Li.
      BACKGROUND: This study aimed to evaluate the inhibitory effects and potential mechanisms of icotinib combined with antiangiogenic drugs on lung adenocarcinoma in vivo.METHODS: A total of 72 mouse xenograft models established with human lung adenocarcinoma cells (HCC827) were randomly divided into six groups, including control, icotinib (Ic), bevacizumab (Bev), recombinant human endostatin (En), Ic + Bev and Ic + En groups. Mouse weights and tumor volumes were measured regularly. Half of the nude mice in each group were sacrificed after 16 days of drug treatment. The remaining animals were observed for another 16 days without drug supply. Immunohistochemical staining was performed to detect microvessel density in tumor heart, liver, brain specimens from the nude mice and Ki67 expression. Differential expression of vascular endothelial growth factor (VEGFA) in tumor tissue specimens was determined by ELISA and Western blot.
    RESULTS: The results showed that the combined drugs inhibited tumor growth more substantially compared with single drugs, without increasing the toxic effects. The antiangiogenesis effect of the combination was better than that of single drug treatment. In addition, both types of targeted drugs and combination medication not only significantly reduced microvessel density in the tumor tissue itself, but also had a certain impact on decreasing microvessel density in the liver. The combination decreased VEGFA and Ki-67 amounts significantly more than icotinib or endostatin as a monotherapy.
    CONCLUSIONS: Icotinib combined with bevacizumab or rh-endostatin has a stronger inhibitory effect on tumor growth than single-target drug in vivo, with no additional side effects.
    Keywords:  bevacizumab; icotinib; microvessel density; recombinant human endostatin (rh-endostatin); vascular endothelial growth factor A (VEGFA)
    DOI:  https://doi.org/10.1111/1759-7714.14261
  40. Front Immunol. 2021 ;12 733921
    Metformin Suppresses Monocyte Immunometabolic Activation by SARS-CoV-2 Spike Protein Subunit 1.
    Theodore J Cory, Russell S Emmons, Johnathan R Yarbro, Kierstin L Davis, Brandt D Pence.
      A hallmark of COVID-19 is a hyperinflammatory state associated with severity. Monocytes undergo metabolic reprogramming and produce inflammatory cytokines when stimulated with SARS-CoV-2. We hypothesized that binding by the viral spike protein mediates this effect, and that drugs which regulate immunometabolism could inhibit the inflammatory response. Monocytes stimulated with recombinant SARS-CoV-2 spike protein subunit 1 showed a dose-dependent increase in glycolytic metabolism associated with production of pro-inflammatory cytokines. This response was dependent on hypoxia-inducible factor-1α, as chetomin inhibited glycolysis and cytokine production. Inhibition of glycolytic metabolism by 2-deoxyglucose (2-DG) or glucose deprivation also inhibited the glycolytic response, and 2-DG strongly suppressed cytokine production. Glucose-deprived monocytes rescued cytokine production by upregulating oxidative phosphorylation, an effect which was not present in 2-DG-treated monocytes due to the known effect of 2-DG on suppressing mitochondrial metabolism. Finally, pre-treatment of monocytes with metformin strongly suppressed spike protein-mediated cytokine production and metabolic reprogramming. Likewise, metformin pre-treatment blocked cytokine induction by SARS-CoV-2 strain WA1/2020 in direct infection experiments. In summary, the SARS-CoV-2 spike protein induces a pro-inflammatory immunometabolic response in monocytes that can be suppressed by metformin, and metformin likewise suppresses inflammatory responses to live SARS-CoV-2. This has potential implications for the treatment of hyperinflammation during COVID-19.
    Keywords:  COVID-19; SARS-CoV-2; immunometabolism; inflammation; monocyte
    DOI:  https://doi.org/10.3389/fimmu.2021.733921
  41. J Biochem. 2021 Nov 25. pii: mvab129. [Epub ahead of print]
    CircRPPH1 promotes cell proliferation, migration and invasion of non-small cell lung cancer (NSCLC) via the PI3K/AKT and JAK2/STAT3 signaling axes.
    Jian-Wen Xiong, Si-Bei Song, Lin-Min Xiong, Chuan-Hui Duan, Qian Song, Dong-Liang Yu, Xiao-Qiang Zhang.
      Non-small cell lung cancer (NSCLC) has markedly increased morbidity and mortality rate worldwide. Circular RNAs (circRNAs) were shown to regulate NSCLC progression. But, the underlying pathways of the circRPPH1-mediated regulation of NSCLC still need further exploration. We evaluated circRPPH1 levels in NSCLC tissues and cell lines via qRT-RCR. Moreover, using ectopic plasmid incorporation and siRNA assays, we analyzed the circRPPH1-mediated regulation of cell proliferation (CP), migration (CM), and invasion (CI) in NSCLC cell lines (H1975 and A549 cells), using CCK-8, colony forming, scratch wound, and transwell assays, respectively. CircRPPH1 levels were remarkably high in the NSCLC tissues and cell lines. The transfection experiments showed that circRPPH1 overexpression was able to promote CP, CM and CI of NSCLC cells, while CP, CM and CI were significantly restrained by the knockdown of circRPPH1. We also displayed that circRPPH1 knockdown suppressed the cell progression via inactivating the PI3K/AKT and JAK2/STAT3 signaling axes. Subsequently, in vivo experiment in nude mice was demonstrated that the inhibition of circRPPH1 could reduce the tumor growth of NSCLC. circRPPH1 may accelerate the growth and metastasis of NSCLC, in culture conditions and in animal models, by stimulating the PI3K/AKT and JAK2/STAT3 signaling axes, thus promoting the development of NSCLC.
    Keywords:  JAK2/STAT3; NSCLC; PI3K/AKT; circRPPH1
    DOI:  https://doi.org/10.1093/jb/mvab129
  42. Support Care Cancer. 2021 Dec 03.
    The prevalence of CT-defined low skeletal muscle mass in patients with metastatic cancer: a cross-sectional multicenter French study (the SCAN study).
    Bruno Raynard, Frederic Pigneur, Mario Di Palma, Elise Deluche, François Goldwasser.
      BACKGROUND: Cachexia, characterized by involuntary muscle mass loss, negatively impacts survival outcomes, treatment tolerability, and functionality in cancer patients. However, there is a limited appreciation of the true prevalence of low muscle mass due to inconsistent diagnostic methods and limited oncologist awareness.METHODS: Twenty-nine French healthcare establishments participated in this cross-sectional study, recruiting patients with those metastatic cancers most frequently encountered in routine practice (colon, breast, kidney, lung, prostate). The primary outcome was low skeletal muscle mass prevalence, as diagnosed by estimating the skeletal mass index (SMI) in the middle of the third-lumbar vertebrae (L3) level via computed tomography (CT). Other objectives included an evaluation of nutritional management, physical activity, and toxicities related to ongoing treatment.
    RESULTS: Seven hundred sixty-six patients (49.9% males) were enrolled with a mean age of 65.0 years. Low muscle mass prevalence was 69.1%. Only one-third of patients with low skeletal muscle mass were receiving nutritional counselling and only 28.4% were under nutritional management (oral supplements, enteral or parenteral nutrition). Physicians highly underdiagnosed those patients identified with low skeletal muscle mass, as defined by the primary objective, by 74.3% and 44.9% in obese and non-obese patients, respectively. Multivariate analyses revealed a lower risk of low skeletal muscle mass for females (OR: 0.22, P < 0.01) and those without brain metastasis (OR: 0.34, P < 0.01). Low skeletal muscle mass patients were more likely to have delayed treatment administration due to toxicity (11.9% versus 6.8%, P = 0.04).
    CONCLUSIONS: There is a critical need to raise awareness of low skeletal muscle mass diagnosis among oncologists, and for improvements in nutritional management and physical therapies of cancer patients to curb potential cachexia. This calls for cross-disciplinary collaborations among oncologists, nutritionists, physiotherapists, and radiologists.
    Keywords:  Cachexia; Low muscle mass; Nutritional support; Skeletal mass index (SMI)
    DOI:  https://doi.org/10.1007/s00520-021-06603-0
  43. PLoS One. 2021 ;16(11): e0260636
    Countering the advert effects of lung cancer on the anticancer potential of dendritic cell populations reinstates sensitivity to anti-PD-1 therapy.
    Julyanne Brassard, Meredith Elizabeth Gill, Emilie Bernatchez, Véronique Desjardins, Joanny Roy, Philippe Joubert, David Marsolais, Marie-Renée Blanchet.
      Lung cancer is the leading cause of cancer-related deaths. While the recent use of immune checkpoint inhibitors significantly improves patient outcomes, responsiveness remains restricted to a small proportion of patients. Conventional dendritic cells (DCs) play a major role in anticancer immunity. In mice, two subpopulations of DCs are found in the lung: DC2s (CD11b+Sirpα+) and DC1s (CD103+XCR1+), the latest specializing in the promotion of anticancer immune responses. However, the impact of lung cancer on DC populations and the consequent influence on the anticancer immune response remain poorly understood. To address this, DC populations were studied in murine models of Lewis Lung Carcinoma (LLC) and melanoma-induced lung metastasis (B16F10). We report that direct exposure to live or dead cancer cells impacts the capacity of DCs to differentiate into CD103+ DC1s, leading to profound alterations in CD103+ DC1 proportions in the lung. In addition, we observed the accumulation of CD103loCD11b+ DCs, which express DC2 markers IRF4 and Sirpα, high levels of T-cell inhibitory molecules PD-L1/2 and the regulatory molecule CD200. Finally, DC1s were injected in combination with an immune checkpoint inhibitor (anti-PD-1) in the B16F10 model of resistance to the anti-PD-1 immune checkpoint therapy; the co-injection restored sensitivity to immunotherapy. Thus, we demonstrate that lung tumor development leads to the accumulation of CD103loCD11b+ DCs with a regulatory potential combined with a reduced proportion of highly-specialized antitumor CD103+ DC1s, which could promote cancer growth. Additionally, promoting an anticancer DC signature could be an interesting therapeutic avenue to increase the efficacy of existing immune checkpoint inhibitors.
    DOI:  https://doi.org/10.1371/journal.pone.0260636
  44. Drug Metab Dispos. 2021 Dec 03. pii: DMD-AR-2021-000581. [Epub ahead of print]
    Pharmacokinetics, Mass Balance, and Metabolism of the Novel URAT1 Inhibitor [14C]HR011303 in Humans: Metabolism is Mediated Predominantly by UDP-glucuronosyltransferase.
    Yuandong Zheng, Hua Zhang, Mengling Liu, Guangze Li, Sheng Ma, Zhe Zhang, Hongda Lin, Yan Zhan, Zhendong Chen, Dafang Zhong, Liyan Miao, Xingxing Diao.
      HR011303, a promising selective URAT1 inhibitor, is currently being studied in a phase Ⅲ clinical trial in China for the treatment of hyperuricemia and gout. In the current study, the pharmacokinetics, mass balance, and metabolism of HR011303 were examined in six healthy Chinese male subjects who received a single oral dose of 10 mg of [14C]HR011303 (80 µCi). The results showed that HR011303 was rapidly absorbed with a median T max = 1.50 h post-dose, and the arithmetic mean t 1/2 of total radioactivity was approximately 24.2 h in plasma. The mean blood-to-plasma radioactivity concentration ratio was 0.66, suggesting the preferential distribution of drug-related components in plasma. At 216 h post-dose, the mean cumulative excreted radioactivity was 91.75% of the dose, including 81.50% in urine and 10.26% in feces. Six metabolites were identified, and the parent drug HR011303 was the most abundant component in plasma and feces, but a minor component in urine. Glucuronidation of the carboxylic acid moiety of HR011303 was the primary metabolic pathway in humans, amounting to 69.63% of the dose (M5, 51.57% of the dose; M5/2, 18.06% of the dose) in the urine; however, it was not detected in plasma. UGT2B7 was responsible for the formation of M5. Overall, after a single oral dose of 10 mg of [14C]HR011303 (80 µCi), HR011303 and its main metabolites were eliminated via renal excretion. The major metabolic pathway was carboxylic acid glucuronidation, which was catalyzed predominantly by UGT2B7. Significance Statement This study determined the absorption and disposition of HR011303, a selective URAT1 inhibitor currently in development for the treatment of hyperuricemia and gout. This work helps to characterize the major metabolic pathways of new URAT inhibitors and identify the absorption and clearance mechanism.
    Keywords:  Mass spectrometry (MS); Radiation; metabolite identification; pharmacokinetic
    DOI:  https://doi.org/10.1124/dmd.121.000581
  45. Transl Lung Cancer Res. 2021 Oct;10(10): 3995-4011
    Deubiquitinase USP5 promotes non-small cell lung cancer cell proliferation by stabilizing cyclin D1.
    Zhiwei Zhang, Zihan Cui, Zhuolin Xie, Chang Li, Chun Xu, Xia Guo, Jie Yu, Tengfei Chen, Francesco Facchinetti, Hanibal Bohnenberger, Tracy L Leong, Yufeng Xie, Xinliang Mao, Jun Zhao.
      Background: Cyclin D1 (CCND1) is overexpressed in non-small cell lung cancer (NSCLC) and contributes to its tumorigenesis and progression. Accumulating evidence shows that ubiquitin-specific protease 5 (USP5), an important member of the USP family, acts as a tumor promoter by deubiquitinating and stabilizing oncoproteins. However, neither the mechanism for dysregulated turnover of CCND1 protein nor the association of CCND1 with USP5 in NSCLC is well understood.Methods: The association of USP5 with CCND1 in human NSCLC cells and clinical tissues was determined by immunoprecipitation/immunoblotting, immunohistochemistry (IHC), and The Cancer Genome Atlas database analyses. The effect of USP5 knockdown or overexpression on NSCLC cell proliferation in vitro was assessed by Cell Counting Kit-8, flow cytometry-based cell cycle, and colony formation assays. The effect of the USP5 inhibitor EOAI3402143 (G9) on NSCLC proliferation in vitro was analyzed by CCK-8 assay. The effect of G9 on NSCLC xenograft tumor growth was also examined in vivo, using athymic BALB/c nude mice.
    Results: USP5 physically bound to CCND1 and decreased its polyubiquitination level, thereby stabilizing CCND1 protein. This USP5-CCND1 axis promoted NSCLC cell proliferation and colony formation. Further, knockdown of USP5 led to CCND1 degradation and cell cycle arrest in NSCLC cells. Importantly, this tumor-suppressive effect elicited by USP5 knockdown in NSCLC cells was validated in vitro and in vivo through chemical inhibition of USP5 activity using G9. Consistently, G9 downregulated the protein levels of CCND1 in NSCLC cells and xenograft tumor tissues. Also, the expression level of USP5 was positively associated with the protein level of CCND1 in human clinical NSCLC tissues.
    Conclusions: This study has provided the first evidence that CCND1 is a novel substrate of USP5. The USP5-CCND1 axis could be a potential target for the treatment of NSCLC.
    Keywords:  Non-small cell lung cancer (NSCLC); cyclin D1 (CCND1); proliferation; ubiquitin-proteasome pathway; ubiquitin-specific protease 5 (USP5)
    DOI:  https://doi.org/10.21037/tlcr-21-767
  46. PLoS One. 2021 ;16(11): e0260500
    Gene expression signatures as candidate biomarkers of response to PD-1 blockade in non-small cell lung cancers.
    Tomoiki Aiba, Chieko Hattori, Jun Sugisaka, Hisashi Shimizu, Hirotaka Ono, Yutaka Domeki, Ryohei Saito, Sachiko Kawana, Yosuke Kawashima, Keisuke Terayama, Yukihiro Toi, Atsushi Nakamura, Shinsuke Yamanda, Yuichiro Kimura, Yutaka Suzuki, Atsushi Niida, Shunichi Sugawara.
      Although anti-PD-1/PD-L1 monotherapy has achieved clinical success in non-small cell lung cancer (NSCLC), definitive predictive biomarkers remain to be elucidated. In this study, we performed whole-transcriptome sequencing of pretreatment tumor tissue samples and pretreatment and on-treatment whole blood samples (WB) samples obtained from a clinically annotated cohort of NSCLC patients (n = 40) treated with nivolumab (anti-PD-1) monotherapy. Using a single-sample gene set enrichment scoring method, we found that the tumors of responders with lung adenocarcinoma (LUAD, n = 20) are inherently immunogenic to promote antitumor immunity, whereas those with lung squamous cell carcinoma (LUSC, n = 18) have a less immunosuppressive tumor microenvironment. These findings suggested that nivolumab may function as a molecular targeted agent in LUAD and as an immunomodulating agent in LUSC. In addition, our study explains why the reliability of PD-L1 expression on tumor cells as a predictive biomarker for the response to nivolumab monotherapy is quite different between LUAD and LUSC.
    DOI:  https://doi.org/10.1371/journal.pone.0260500
  47. Virchows Arch. 2021 Nov 30.
    Distinct properties of pure- and mixed-type high-grade fetal lung adenocarcinomas by genetic profiling and transcription factor expression.
    Satsuki Kishikawa, Takuo Hayashi, Tsuyoshi Saito, Kazuya Takamochi, Keita Sasa, Yoshiyuki Suehara, Fumiyuki Takahashi, Noriko Sasahara, Shinji Kohsaka, Kenji Suzuki, Takashi Yao.
      The clinicopathological differences among high-grade fetal lung adenocarcinomas completely comprising tumor cells that resemble fetal lung epithelium (pure type) and those with fetal lung-like components admixed with conventional adenocarcinoma cells (mixed type) remain undetermined. Here, we examined the clinicopathological, immunohistochemical, and molecular features of 11 lung adenocarcinomas with fetal lung-like morphology among 3895 consecutive cases of primary lung cancer based on the expression pattern of transcription factors. According to the current WHO classification, two cases (0.05%) were categorized as low-grade fetal adenocarcinoma, two cases (0.05%) were pure-type high-grade fetal adenocarcinoma, five cases (0.1%) were mixed-type high-grade fetal adenocarcinoma, and the remaining two cases (0.05%) were lung adenocarcinoma with high-grade fetal features (fetal lung-like morphology occupied less than 50%). CTNNB1 mutations were exclusively identified in low-grade fetal adenocarcinomas. In contrast, mixed-type high-grade fetal adenocarcinoma or lung adenocarcinoma with high-grade fetal features frequently harbored mitogenic drivers including EGFR mutations. Furthermore, almost all tumor cells expressed CDX2 and HNF4α in both cases of pure-type high-grade fetal lung adenocarcinoma, but lacked TTF-1 positivity. In contrast, TTF-1 was frequently expressed in mixed-type high-grade fetal lung adenocarcinoma and in lung adenocarcinoma with high-grade fetal features. Our data suggest similar prevalence of low-grade fetal lung adenocarcinoma and pure-type high-grade fetal lung adenocarcinoma, and indicate that pure- and mixed-type high-grade fetal lung adenocarcinomas are distinct, with the former akin to low-grade fetal adenocarcinoma with respect to purely embryonic morphology and absence of common lung adenocarcinoma mitogenic drivers, and the latter being genetically and transcriptionally related to conventional lung adenocarcinoma.
    Keywords:  CDX2; Fetal lung adenocarcinoma; High-grade fetal lung adenocarcinoma; TTF-1
    DOI:  https://doi.org/10.1007/s00428-021-03247-7
  48. EClinicalMedicine. 2021 Dec;42 101187
    Efficacy and safety of MIL60 compared with bevacizumab in advanced or recurrent non-squamous non-small cell lung cancer: a phase 3 randomized, double-blind study.
    Rui Wan, Xiaorong Dong, Qun Chen, Yan Yu, Shujun Yang, Xiaochun Zhang, Guojun Zhang, Yueyin Pan, Sanyuan Sun, Chengzhi Zhou, Wei Hong, Hui Zhao, Lei Yang, Linian Huang, Rong Wu, Aimin Zang, Rui Ma, Lin Wu, Dongqing Lv, Xiuhua Fu, Jianguo Han, Wenxin Li, Jianchun Duan, Kai Wang, Ou Jiang, Yinglan Chen, Zhongliang Guo, Hongjun Gao, Juyi Wen, Shubin Wang, Enfeng Zhao, Gaofeng Li, Lu Yue, Li Liang, Aiping Zeng, Xiaoshan Wang, Yuxi Zhu, Hongming Pan, Zhaoxia Dai, Weineng Feng, Guofang Zhao, Chuan Lin, Chong Li, Na Li, Yangyi Bao, Yinyin Li, Yanjun Su, Min Zhao, Haohui Fang, Yulong Zhu, Yu Zhang, Lieming Ding, Yang Wang, Xiaobin Yuan, Jie Wang.
      Background: We compared the efficacy, safety, and immunogenicity of MIL60 with reference bevacizumab as first-line treatment in patients with advanced or recurrent non-squamous non-small cell lung cancer (NSCLC) in this phase 3, randomized, double-blind study.Methods: Patients with untreated advanced or recurrent NSCLC were randomized (1:1 ratio) to receive either MIL60 or bevacizumab in combination with paclitaxel/carboplatin. Patients with non-progressive disease continued maintenance single-agent MIL60 until disease progression, or intolerable toxicity. The primary endpoint was the 12-week objective response rates (ORR12) by independent review committee (IRC) using RECIST 1.1. Bioequivalence was established if the ORR ratio located between 0.75 and 1/0.75. The trial was registered with clinicaltrials.gov (NCT03196986).
    Findings: Between Aug 23, 2017, and May 8, 2019, 517 patients were randomly assigned to MIL60 group (n=257) and bevacizumab group (n=260). In the full analysis set (FAS) population including all randomized and evaluable patients who received at least one dose of MIL60 or bevacizumab, the ORR12 in MIL60 group and bevacizumab group were 48.6% and 43.1%, respectively. The ORR ratio of these two groups were 1.14 (90% CI 0.97-1.33), which fell within the pre-specified equivalence boundaries (0.75-1/0.75). The median DOR was 5.7 months (95% CI 4.5-6.2) for MIL60 and 5.6 months (95% CI 4.3-6.4) for bevacizumab. No significant difference was noted in median PFS (7.2 vs. 8.1 months; HR 1.01, 95% CI 0.78-1.30, p=0.9606) and OS (19.3 vs. 16.3 months; HR 0.81, 95% CI 0.64-1.02, p=0.0755). Safety and tolerability profiles were similar between the two groups. No patient detected positive for Anti-drug antibody (ADA).
    Interpretation: The efficacy, safety and immunogenicity of MIL60 were similar with bevacizumab, providing an alternative treatment option for advanced or recurrent non-squamous NSCLC.
    Funding: This study was sponsored by Betta Pharmaceutical Co., Ltd.
    Keywords:  MIL60; bevacizumab; biosimilar; equivalence; non-squamous NSCLC
    DOI:  https://doi.org/10.1016/j.eclinm.2021.101187
  49. Cancer Cell Int. 2021 Nov 27. 21(1): 628
    PESV represses non-small cell lung cancer cell malignancy through circ_0016760 under hypoxia.
    Hong Zhang, Haojian Zhang, Jiye Zhu, Huan Liu, Qin Zhou.
      BACKGROUND: Non-small cell lung cancer (NSCLC) accounts for more than 80% of lung cancers, which is the most common malignant tumor worldwide. Polypeptide extract from scorpion venom (PESV) has been reported to inhibit NSCLC process. The present study aims to reveal the roles of PESV in NSCLC progression under hypoxia and the inner mechanism.METHODS: The expression levels of circular RNA 0016760 (circ_0016760) and microRNA-29b (miR-29b) were detected by quantitative real-time polymerase chain reaction (qRT-PCR). Protein expression was determined by western blot and immunohistochemistry assays. Cell migration, invasion, proliferation and tube formation were investigated by transwell, cell colony formation, 3-(4,5-Dimethylthazol-2-yl)-2,5-diphenyltetrazolium bromide and tube formation assays. The impacts between PESV and circ_0016760 overexpression on tumor growth in vivo were investigated by in vivo tumor formation assay.
    RESULTS: Circ_0016760 expression was dramatically upregulated in NSCLC tissues and cells, compared with adjacent lung tissues and cells, respectively. PESV treatment downregulated circ_0016760 expression. Circ_0016760 silencing or PESV treatment repressed cell migration, invasion, proliferation and tube formation under hypoxia in NSCLC cells. Circ_0016760 overexpression restored the effects of PESV treatment on NSCLC process under hypoxia. Additionally, circ_0016760 acted as a sponge of miR-29b, and miR-29b bound to HIF1A. Meanwhile, miR-29b inhibitor impaired the influences of circ_0016760 knockdown on NSCLC process under hypoxia. Further, ectopic circ_0016760 expression restrained the effects of PESV exposure on tumor formation in vivo.
    CONCLUSION: Circ_0016760 overexpression counteracted PESV-induced repression of NSCLC cell malignancy and angiogenesis under hypoxia through miR-29b/HIF1A axis.
    Keywords:  HIF1A; NSCLC; PESV; circ_0016760; miR-29b
    DOI:  https://doi.org/10.1186/s12935-021-02336-6
  50. Crit Rev Oncol Hematol. 2021 Nov 24. pii: S1040-8428(21)00332-2. [Epub ahead of print]169 103545
    Targeting the metabolism of cancer stem cells by energy disruptor molecules.
    Tahere Dadgar, Nasim Ebrahimi, Amir Reza Gholipour, Maryam Akbari, Leila Khani, Amirhossein Ahmadi, Michael R Hamblin.
      Cancer stem cells (CSCs) have been identified in various tumor types. CSCs are believed to contribute to tumor metastasis and resistance to conventional therapy. So targeting these cells could be an effective strategy to eliminate tumors and a promising new type of cancer treatment. Alterations in metabolism play an essential role in CSC biology and their resistance to treatment. The metabolic properties pathways in CSCs are different from normal cells, and to some extent, are different from regular tumor cells. Interestingly, CSCs can use other nutrients for their metabolism and growth. The different metabolism causes increased sensitivity of CSCs to agents that disrupt cellular homeostasis. Compounds that interfere with the central metabolic pathways are known as energy disruptors and can reduce CSC survival. This review highlights the differences between regular cancer cells and CSC metabolism and discusses the action mechanisms of energy disruptors at the cellular and molecular levels.
    Keywords:  Cancer; Cancer stem cells; Energy disruptors; Metabolism
    DOI:  https://doi.org/10.1016/j.critrevonc.2021.103545
  51. Comput Struct Biotechnol J. 2021 ;19 5920-5930
    Gut microbiome signatures distinguish type 2 diabetes mellitus from non-alcoholic fatty liver disease.
    Jiyeon Si, Giljae Lee, Hyun Ju You, Sae Kyung Joo, Dong Hyeon Lee, Bon Jeong Ku, Seoyeon Park, Won Kim, GwangPyo Ko.
      Non-alcoholic fatty liver disease (NAFLD) is closely associated with type 2 diabetes mellitus (T2D), and these two metabolic diseases demonstrate bidirectional influences. The identification of microbiome profiles that are specific to liver injury or impaired glucose metabolism may assist understanding of the role of the gut microbiota in the relationship between NAFLD and T2D. Here, we studied a biopsy-proven Asian NAFLD cohort (n = 329; 187 participants with NAFLD, 101 with NAFLD and T2D, and 41 with neither) and identified Enterobacter, Romboutsia, and Clostridium sensu stricto as the principal taxa associated with the severity of NAFLD and T2D, whereas Ruminococcus and Megamonas were specific to NAFLD. In particular, the taxa that were associated with both severe liver pathology and T2D were also significantly associated with markers of diabetes, such as fasting blood glucose and Hb1Ac. Enterotype analysis demonstrated that participants with NAFLD had a significantly higher proportion of Bacteroides and a lower proportion of Ruminococcus than a Korean healthy twin cohort (n = 756). However, T2D could not be clearly distinguished from NAFLD. Analysis of an independent T2D cohort (n = 185) permitted us to validate the T2D-specific bacterial signature identified in the NAFLD cohort. Functional inference analysis revealed that endotoxin biosynthesis pathways were significantly enriched in participants with NAFLD and T2D, compared with those with NAFLD alone. These findings may assist with the development of effective therapeutic approaches for metabolic diseases that are associated with specific bacterial signatures.
    Keywords:  ALT, alanine aminotransferase; BMI, body mass index; Biomarker; Enterotype; FBS, fasting blood sugar; FDR, false discovery rate; FLI, fatty liver index; Gut microbiome; HbA1c, glycosylated hemoglobin; LDL, low-density lipoprotein; LPS, lipopolysaccharide; MaAsLin2, microbiome multivariable association with linear models 2; NAFL, non-alcoholic fatty liver; NAFLD, non-alcoholic fatty liver disease; NASH, non-alcoholic steatohepatitis; NASH-CRN, non-alcoholic steatohepatitis clinical research network; Non-alcoholic fatty liver disease (NAFLD); PICRUSt2, phylogenetic investigation of communities by reconstruction of unobserved states 2; T2D, type 2 diabetes mellitus; Type 2 diabetes mellitus
    DOI:  https://doi.org/10.1016/j.csbj.2021.10.032
  52. Future Oncol. 2021 Nov 29.
    Plasma SNORD83A as a potential biomarker for early diagnosis of non-small-cell lung cancer.
    Kangyu Wang, Xingguo Song, Xinyi Li, Zhijun Zhang, Li Xie, Xianrang Song.
      Aim: This study aimed to access the efficacy of plasma small nucleolar RNAs in early diagnosis of non-small-cell lung cancer (NSCLC). Methods: SNORD83A was selected based on databases and further verified in 48 paired formalin-fixed, paraffin-embedded tissues, as well as in plasma from 150 NSCLC patients and 150 healthy donors. The diagnostic efficiency of plasma SNORD83A, as well as in combination with carcinoembryonic antigen, was determined by receiver operating characteristic analysis. Results: SNORD83A was significantly increased not only in tissues but also in plasma from NSCLC patients compared with those from healthy donors. Plasma SNORD83A was able to act as a diagnostic biomarker for NSCLC. The diagnostic efficiency of carcinoembryonic antigen was also significantly elevated for early-stage NSCLC when combined with SNORD83A. Conclusion: SNORD83A can serve as a diagnostic biomarker for NSCLC.
    Keywords:  SNORD83A; biomarker; diagnosis; non-small-cell lung cancer; snoRNAs
    DOI:  https://doi.org/10.2217/fon-2021-1278
  53. Technol Cancer Res Treat. 2021 Jan-Dec;20:20 15330338211039948
    Aberrant Alteration of Circulating Lymphocyte Subsets in Small Cell Lung Cancer Patients Treated with Radiotherapy.
    Hui Li, Hong Yu, Shaowei Lan, Dandan Zhao, Yan Liu, Ying Cheng.
      Purpose: The role of different circulating lymphocyte subsets, as well as their correlation with clinical characteristics of small cell lung cancer patients have not yet been fully understood. This study aims to evaluate the influence of the fluctuating absolute numbers of lymphocyte subpopulations in peripheral blood of patients with small cell lung cancer. Methods: The absolute counts and percentages of lymphocyte subsets in peripheral blood of 329 patients with small cell lung cancer were retrospectively analyzed. The numbers of CD3+, CD3+CD4+, and CD3+CD8+ T lymphocytes, CD3-CD19+ B lymphocytes, and CD3-CD16+CD56+ NK cells were evaluated by flow cytometry. Their relationship with the patients' clinical characteristics were statistically evaluated. Results: The CD4/CD8 values derived from the absolute number and percentage of CD3+CD4+ cells divided by CD3+CD8+ cells were identical (1.86  ±  0.99). There was no association between any of the lymphocyte subsets levels and age/sex of the 329 patients with small cell lung cancer. The patients with advanced stage had a reduction in CD3+ and CD3+CD4+ T cell counts and a decreased CD4/CD8 ratio. The levels of CD3+CD4+ T cells, CD3-CD19+ B cells, CD3-CD16+CD56+ NK cells, and CD4/CD8 ratio were associated with advanced tumor-node-metastasis stage. Patients who had undergone radiotherapy were characterized by lymphopenia with lower numbers of CD3+, CD3+CD4+, CD3+CD8+ T lymphocyte, B lymphocyte, NK cell, and CD4/CD8 ratio. The evaluation of individual CD4/CD8 ratio should be combined with other clinical parameters. Conclusions: Patients with small cell lung cancer have altered lymphocyte homeostasis. Lymphopenia was a long-lasting feature of the enrolled patients who were treated with radiotherapy. The available lymphocyte subsets levels might be used to manage the clinical treatment scheme.
    Keywords:  immune status; lymphocyte subsets; lymphopenia; radiotherapy; small cell lung cancer
    DOI:  https://doi.org/10.1177/15330338211039948
  54. Thorax. 2021 Dec 01. pii: thoraxjnl-2021-217001. [Epub ahead of print]
    Tumour-infiltrating bystander CD8+ T cells activated by IL-15 contribute to tumour control in non-small cell lung cancer.
    Galam Leem, Minwoo Jeon, Kun Woo Kim, Seongju Jeong, Seong Jin Choi, Yong Joon Lee, Eui-Soon Kim, Jae-Ik Lee, Seung Yeon Ha, Su-Hyung Park, Hyo Sup Shim, Jin Gu Lee, Shin Myung Kang, Eui-Cheol Shin.
      BACKGROUND: Tumour-unrelated, virus-specific bystander CD8+ T cells were recently shown to be abundant among tumour-infiltrating lymphocytes (TILs). However, their roles in tumour immunity have not been elucidated yet.METHODS: We studied the characteristics of bystander CD8+ TILs from non-small cell lung cancer (NSCLC) tissues (N=66) and their activation by interleukin (IL)-15 to repurpose them for tumour immunotherapy.
    RESULTS: We show that bystander CD8+ TILs specific to various viruses are present in human NSCLC tissues. We stimulated CD8+ TILs ex vivo using IL-15 without cognate antigens and found that IL-15 treatment upregulated NKG2D expression on CD8+ TILs, resulting in NKG2D-dependent production of interferon (IFN)-γ (p=0.0006). Finally, we tested whether IL-15 treatment can control tumour growth in a murine NSCLC model with or without a history of murine cytomegalovirus (MCMV) infection. IL-15 treatment reduced the number of tumour nodules in the lung only in mice with MCMV infection (p=0.0037). We confirmed that MCMV-specific bystander CD8+ TILs produced interferon (IFN)-γ after IL-15 treatment, and that IL-15 treatment in MCMV-infected mice upregulated tumour necrosis factor-α and IFN-γ responsive genes in tumour microenvironment.
    CONCLUSION: Thus, the study demonstrates that bystander CD8+ TILs can be repurposed by IL-15 for tumour immunotherapy.
    Keywords:  lung cancer; non-small cell lung cancer
    DOI:  https://doi.org/10.1136/thoraxjnl-2021-217001
  55. Nutr Metab (Lond). 2021 Nov 27. 18(1): 102
    Prevalence and prognostic significance of malnutrition in diabetic patients with coronary artery disease: a cohort study.
    Wen Wei, Lingyu Zhang, Guode Li, Zhidong Huang, Jin Liu, Zhihuang Wu, Yuanying Wu, Jinrong Lin, Yunhan Zhang, Yaren Yu, Haozhang Huang, Qiang Li, Bo Wang, Yong Liu, Mei Tu, Hong Chen, Shiqun Chen.
      BACKGROUND: Malnutrition is associated with poor prognosis in cardiovascular disease patients or in diabetic patients. However, the relationship between malnutrition and clinical outcomes in diabetic patients with coronary artery disease (CAD) is not well known. The aim of this study is to report the prevalence and prognostic consequences of malnutrition in diabetic patients with CAD.METHODS: In this retrospective observational study, the Controlling Nutritional Status (CONUT) score applied to 12,898 consecutive diabetic patients with CAD. The association between malnutrition and long-term all-cause mortality was examined using Cox proportional hazards regression analysis.
    RESULTS: According to CONUT score, 60.5% patients suffered from malnutrition; 46.4%, 13.2%, and 0.9% patients had mild, moderate, and severe malnutrition, respectively. During a median follow-up of 4.88 (2.83-7.51) years, 1973 (15.3%) patients died. After adjustment for confounders, malnutrition was associated with significantly increased risk for long-term all-cause mortality (adjusted hazard ratio for mild malnutrition and moderate to severe malnutrition, respectively: 1.38 [95% confidence interval (CI) 1.07-1.77]; P value = 0.012 and 1.63 [95% CI 1.18-2.24]; P value = 0.003). A similar association was observed around subgroups.
    CONCLUSIONS: Malnutrition is common in diabetic patients with CAD and is strongly associated with increased mortality. It is necessary to adequately assess the nutritional status and take the effective nutritional guidance to improve the prognosis of diabetic patients with CAD.
    Keywords:  Coronary artery disease; Diabetic; Malnutrition; Prevalence; Prognosis
    DOI:  https://doi.org/10.1186/s12986-021-00626-4
  56. Oncoimmunology. 2021 ;10(1): 2006893
    Keratinocyte differentiation antigen-specific T cells in immune checkpoint inhibitor-treated NSCLC patients are associated with improved survival.
    Fiamma Berner, Rebekka Niederer, Jolien J Luimstra, Oltin Tiberiu Pop, Ann-Kristin Jochum, Mette-Triin Purde, Omar Hasan Ali, David Bomze, Jens Bauer, Lena Katharina Freudenmann, Ana Marcu, Eva-Maria Wolfschmitt, Sebastian Haen, Thorben Gross, Marissa Lisa Dubbelaar, Marie-Therese Abdou, Petra Baumgaertner, Christina Appenzeller, Caroline Cicin-Sain, Tobias Lenz, Daniel E Speiser, Burkhard Ludewig, Christoph Driessen, Markus Jörger, Martin Früh, Wolfram Jochum, Antonio Cozzio, Hans-Georg Rammensee, Juliane Walz, Jacques Neefjes, Lukas Flatz.
      Immune checkpoint inhibitors (ICIs) have improved the survival of patients with non-small cell lung cancer (NSCLC) by reinvigorating tumor-specific T cell responses. However, the specificity of such T cells and the human leukocyte antigen (HLA)-associated epitopes recognized, remain elusive. In this study, we identified NSCLC T cell epitopes of recently described NSCLC-associated antigens, termed keratinocyte differentiation antigens. Epitopes of these antigens were presented by HLA-A 03:01 and HLA-C 04:01 and were associated with responses to ICI therapy. Patients with CD8+ T cell responses to these epitopes had improved overall and progression-free survival. T cells specific for such epitopes could eliminate HLA class I-matched NSCLC cells ex vivo and were enriched in patient lung tumors. The identification of novel lung cancer HLA-associated epitopes that correlate with improved ICI-dependent treatment outcomes suggests that keratinocyte-specific proteins are important tumor-associated antigens in NSCLC. These findings improve our understanding of the mechanisms of ICI therapy and may help support the development of vaccination strategies to improve ICI-based treatment of these tumors.
    Keywords:  Immune checkpoint inhibitor therapy; NSCLC; autoimmune toxicity; tumor-associated antigen
    DOI:  https://doi.org/10.1080/2162402X.2021.2006893
  57. Cancer Biol Med. 2021 Dec 01. pii: j.issn.2095-3941.2021.0178. [Epub ahead of print]
    BLM helicase inhibition synergizes with PARP inhibition to improve the radiosensitivity of olaparib resistant non-small cell lung cancer cells by inhibiting homologous recombination repair.
    Yangyang Kong, Chang Xu, Xiaohui Sun, Hao Sun, Xiaotong Zhao, Ningning He, Kaihua Ji, Qin Wang, Liqing Du, Jinhan Wang, Manman Zhang, Yang Liu, Yan Wang, Qiang Liu.
      OBJECTIVE: We aimed to investigate the radiosensitizing efficacy of the poly-ADP-ribose polymerase (PARP) inhibitor, olaparib, and the Bloom syndrome protein (BLM) helicase inhibitor, ML216, in non-small cell lung cancer (NSCLC) cells.METHODS: Radiosensitization of NSCLC cells was assessed by colony formation and tumor growth assays. Mechanistically, the effects of ML216, olaparib, and radiation on cell and tumor proliferation, DNA damage, cell cycle, apoptosis, homologous recombination (HR) repair, and non-homologous end joining (NHEJ) repair activity were determined.
    RESULTS: Both olaparib and ML216 enhanced the radiosensitivities of olaparib-sensitive H460 and H1299 cells, which was seen as decreased surviving fractions and Rad51 foci, increased total DNA damage, and γH2AX and 53BP1 foci (P < 0.05). The expressions of HR repair proteins were remarkably decreased in olaparib-treated H460 and H1299 cells after irradiation (P < 0.05), while olaparib combined with ML216 exerted a synergistic radiosensitization effect on olaparib-resistant A549 cells. In addition to increases of double strand break (DSB) damage and decreases of Rad51 foci, olaparib combined with ML216 also increased pDNA-PKcs (S2056) foci, abrogated G2 cell cycle arrest, and induced apoptosis in A549 lung cancer after irradiation in vitro and in vivo (P < 0.05). Moreover, Western blot showed that olaparib combined with ML216 and irradiation inhibited HR repair, promoted NHEJ repair, and inactivated cell cycle checkpoint signals both in vitro and in vivo (P < 0.05).
    CONCLUSIONS: Taken together, these results showed the efficacy of PARP and BLM helicase inhibitors for radiosensitizing NSCLC cells, and supported the model that BLM inhibition sensitizes cells to PARP inhibitor-mediated radiosensitization, as well as providing the basis for the potential clinical development of this combination for tumors intrinsically resistant to PARP inhibitors and radiotherapy.
    Keywords:  BLM; NSCLC; PARP; homologous recombination repair; radiosensitization
    DOI:  https://doi.org/10.20892/j.issn.2095-3941.2021.0178
  58. Thorac Cancer. 2021 Dec 01.
    CircRNA circ_0008037 facilitates tumor growth and the Warburg effect via upregulating NUCKS1 by binding to miR-433-3p in non-small cell lung cancer.
    Jia Yu, Haining Zhang, Chunsheng Zhao, Guanghui Li, Yingying Zhang, Yang Sun.
      BACKGROUND: Circular RNAs (circRNAs) participate in the genesis and progression of tumors, including non-small cell lung cancer (NSCLC). At present, the role and regulatory mechanisms of circRNAs in NSCLC have not been fully elucidated. The aim of this study was to explore the role and regulatory mechanism of circRNA hsa_circ_0008037 (circ_0008037) in NSCLC.METHODS: Expression of circ_0008037 in NSCLC tissues and cells was detected by quantitative real-time polymerase chain reaction (RT-qPCR). Loss-of-function experiments were performed to analyze the influence of circ_0008037 knockdown on proliferation, migration, invasion, and the Warburg effect of NSCLC cells. Western blotting was utilized for protein analysis. The regulatory mechanism of circ_0008037 was surveyed by bioinformatics analysis, RNA pulldown assay, and dual-luciferase reporter assay. Xenograft assay was used to validate the oncogenicity of circ_0008037 in NSCLC in vivo.
    RESULTS: Circ_0008037 was upregulated in NSCLC tissues and cells. Circ_0008037 downregulation reduced tumor growth in vivo and repressed proliferation, migration, invasion, and decreased the Warburg effect of NSCLC cells in vitro. Mechanically, circ_0008037 regulated nuclear ubiquitous casein kinase and cyclin-dependent kinase substrate 1 (NUCKS1) expression via sponging miR-433-3p. Furthermore, MiR-433-3p inhibitor reversed the inhibiting influence of circ_0008037 silencing on proliferation, migration, invasion, and the Warburg effect of NSCLC cells. Also, NUCKS1 elevation overturned the repressive influence of miR-433-3p mimic on proliferation, migration, invasion, and the Warburg effect of NSCLC cells.
    CONCLUSION: Circ_0008037 accelerated tumor growth and elevated the Warburg effect via regulating NUCKS1 expression by adsorbing miR-433-3p, providing an underlying target for NSCLC treatment.
    Keywords:  NSCLC; NUCKS1; circ_0008037; miR-433-3p
    DOI:  https://doi.org/10.1111/1759-7714.14235
  59. EMBO Mol Med. 2021 Nov 30. e14296
    A new ALK inhibitor overcomes resistance to first- and second-generation inhibitors in NSCLC.
    Yue Lu, Zhenzhen Fan, Su-Jie Zhu, Xiaoxing Huang, Zhongji Zhuang, Yunzhan Li, Zhou Deng, Lei Gao, Xuehui Hong, Ting Zhang, Li Li, Xihuan Sun, Wei Huang, Jingfang Zhang, Yan Liu, Baoding Zhang, Jie Jiang, Fu Gui, Zheng Wang, Qiyuan Li, Siyang Song, Xin Huang, Qiao Wu, Lanfen Chen, Dawang Zhou, Jianming Zhang, Cai-Hong Yun, Liang Chen, Xianming Deng.
      More than 60% of nonsmall cell lung cancer (NSCLC) patients show a positive response to the first ALK inhibitor, crizotinib, which has been used as the standard treatment for newly diagnosed patients with ALK rearrangement. However, most patients inevitably develop crizotinib resistance due to acquired secondary mutations in the ALK kinase domain, such as the gatekeeper mutation L1196M and the most refractory mutation, G1202R. Here, we develop XMU-MP-5 as a new-generation ALK inhibitor to overcome crizotinib resistance mutations, including L1196M and G1202R. XMU-MP-5 blocks ALK signaling pathways and inhibits the proliferation of cells harboring either wild-type or mutant EML4-ALK in vitro and suppresses tumor growth in xenograft mouse models in vivo. Structural analysis provides insights into the mode of action of XMU-MP-5. In addition, XMU-MP-5 induces significant regression of lung tumors in two genetically engineered mouse (GEM) models, further demonstrating its pharmacological efficacy and potential for clinical application. These preclinical data support XMU-MP-5 as a novel selective ALK inhibitor with high potency and selectivity. XMU-MP-5 holds great promise as a new therapeutic against clinically relevant secondary ALK mutations.
    Keywords:  ALK inhibitor; EML4-ALK; acquired resistance mutations; crizotinib; nonsmall cell lung cancer
    DOI:  https://doi.org/10.15252/emmm.202114296
  60. Transl Lung Cancer Res. 2021 Oct;10(10): 3983-3994
    Prognostic significance of postoperative longitudinal change of serum carcinoembryonic antigen level in patients with stage I lung adenocarcinoma completely resected by single-port video-assisted thoracic surgery: a retrospective study.
    Hao Chen, Yan Jiang, Keyi Jia, Kaixuan Zhang, Natsumi Matsuura, Jin Yong Jeong, Bo Su, Xiao Zhou.
      Background: Patients with stage I lung adenocarcinoma (LUAD) have varying postoperative prognosis. This study aimed to investigate the prognostic significance of postoperative longitudinal change of serum carcinoembryonic antigen (CEA) level in patients with stage I LUAD.Methods: The study cohort comprised 241 patients with stage I LUAD completely resected with single-port video-assisted thoracic surgery (VATS). The patients were categorized into 4 groups according to the postoperative longitudinal change of serum CEA levels measured in the third and sixth months after surgery: the NN group (continuously normal), HN group (increase first and then decrease), NH group (decrease first and then increase), and HH group (continuously high). Recurrence-free survival (RFS) was analyzed by the Kaplan-Meier method and compared by log-rank test. A nomogram was developed to predict recurrence in the stage I LUAD patients.
    Results: In univariate analysis, differentiation (P<0.001), visceral pleural invasion (VPI) (P=0.025), tumor diameter (P<0.001), tumor-node-metastasis (TNM) stage (P=0.008), preoperative CEA levels (≥10.0 vs. <10.0 ng/mL, P<0.001), and postoperative CEA grouping (NH/HH vs. NN/HN, P<0.001) were significant prognostic factors for stage I LUAD patients. Multivariate analysis showed that tumor diameter (P=0.009) and postoperative CEA grouping (P<0.001) were considered to be independent prognostic factors of postoperative recurrence of stage I LUAD. Tumor diameter (≥20 mm) and postoperative CEA (NH/HH vs. NN/HN) were associated with worse RFS. Receiver operating characteristic (ROC) curve analysis showed that postoperative CEA (NH/HH vs. NN/HN) have high sensitivity (64.7%) and specificity (83.2%) for early prediction of postoperative recurrence of stage I LUAD. The area under curve (AUC) value was 0.745. The nomogram based on multivariate Cox regression had a concordance index (value of 0.789). The calibration plot showed that the predicted probabilities closely matched the observed probabilities.
    Conclusions: Longitudinal change in serum CEA level after surgery was found to be an independent unfavorable prognostic factor in completely resected stage I LUAD patients. The NH group and HH group were significantly associated with worse RFS. A nomogram was established to predict the postoperative recurrence of patients with stage I LUAD.
    Keywords:  Lung adenocarcinoma (LUAD); carcinoembryonic antigen (CEA); longitudinal; minimal residual disease (MRD); postoperative
    DOI:  https://doi.org/10.21037/tlcr-21-833
  61. Rev Endocr Metab Disord. 2021 Dec 02.
    Metabolomic signatures after bariatric surgery - a systematic review.
    Matilde Vaz, Sofia S Pereira, Mariana P Monteiro.
      Metabolomics emerged as an important tool to gain insights on how the body responds to therapeutic interventions. Bariatric surgery is the most effective treatment for severe obesity and obesity-related co-morbidities. Our aim was to conduct a systematic review of the available data on metabolomics profiles that characterize patients submitted to different bariatric surgery procedures, which could be useful to predict clinical outcomes including weight loss and type 2 diabetes remission. For that, the Preferred Reporting Items for Systematic Reviews and Meta-Analyses - PRISMA guidelines were followed. Data from forty-seven original study reports addressing metabolomics profiles induced by bariatric surgery that met eligibility criteria were compiled and summarized. Amino acids, lipids, energy-related and gut microbiota-related were the metabolite classes most influenced by bariatric surgery. Among these, higher pre-operative levels of specific lipids including phospholipids, long-chain fatty acids and bile acids were associated with post-operative T2D remission. As conclusion, metabolite profiling could become a useful tool to predict long term response to different bariatric surgery procedures, allowing more personalized interventions and improved healthcare resources allocation.
    Keywords:  Bariatric surgery; Metabolomics; Type 2 diabetes remission; Weight loss
    DOI:  https://doi.org/10.1007/s11154-021-09695-5
  62. Chemosphere. 2021 Nov 30. pii: S0045-6535(21)03597-9. [Epub ahead of print] 133125
    Metabolomic analysis and oxidative stress response reveals the toxicity in Escherichia coli induced by organophosphate flame retardants tris(2-chloroethyl) phosphate and triphenyl phosphate.
    Xiaolong Yu, Xu Jin, Jin Tang, Nan Wang, Yuanyuan Yu, Rongrong Sun, Fucai Deng, Chudan Huang, Jianteng Sun, Lizhong Zhu.
      Organophosphate flame retardants (OPFRs) are emerging environmental pollutants that are increasingly being used in consumer commodities. The adverse effects on biota induced by tris(2-chloroethyl) phosphate (TCEP) and triphenyl phosphate (TPHP) have become a growing concern. Unfortunately, toxic mechanisms at the molecular level for OPFRs in organisms are still lacking. Herein, Escherichia coli (E.coli) was exposed to TCEP and TPHP for 24 and 48 h to reveal oxidative stress response and molecular toxicity mechanisms. The results indicated that promotion of ROS overload occurred at higher dosages groups. The levels of SOD and CAT were significantly elevated along with the increase of MDA attributed to lipid peroxidation. Additionally, apoptosis rates increased, accompanied by a decline in membrane potential and Na+/K+-ATPase and Ca2+/Mg2+-ATPase contents, signifying that E. coli cytotoxicity induced by TCEP and TPHP was mediated by oxidative stress. Based on metabolomic analysis, different metabolic pathways were disrupted, including glycolysis/gluconeogenesis, pentose phosphate metabolism, purine metabolism, glutathione metabolism, amino acid biosynthesis, butanoate metabolism, alanine and aspartate metabolism. Most differentially expressed metabolites were downregulated, indicating an inhibitory effect on metabolic functions and key metabolic pathways. These findings generated new insights into the potential environmental risks of OPFRs in aquatic organisms.
    Keywords:  Apoptosis; Differential metabolites; Metabolomics; Organophosphorus flame retardants; Oxidative stress response
    DOI:  https://doi.org/10.1016/j.chemosphere.2021.133125
  63. Cancer Immunol Immunother. 2021 Nov 28.
    Baseline tumour size is an independent prognostic factor for overall survival in PD-L1 ≥ 50% non-small cell lung cancer patients treated with first-line pembrolizumab.
    Mathilde Bureau, Thierry Chatellier, Tanguy Perennec, Thomas Goronflot, Charlotte Greilsamer, Anne-Laure Chene, Raafet Affi, Eric Frampas, Jaafar Bennouna, Elvire Pons-Tostivint.
      BACKGROUND: Advanced non-small cell lung cancer (NSCLC) with a PD-L1 tumour proportion score ≥ 50% can be treated with pembrolizumab alone. Our aim was to assess the impact of baseline tumour size (BTS) on overall survival (OS) in NSCLC patients treated with pembrolizumab versus chemotherapy.METHODS: This retrospective, multicentre study included all patients with untreated advanced NSCLC receiving either pembrolizumab (PD-L1 ≥ 50%) or platinum-based chemotherapy (any PD-L1). The primary endpoint was the impact of BTS (defined as the sum of the dimensions of baseline target lesions according to RECIST v1.1 criteria) on OS.
    RESULTS: Between 09-2016 and 06-2020, 188 patients were included, 96 in the pembrolizumab (P-group) and 92 in the chemotherapy group (CT-group). The median follow-up was 26.9 months (range 0.13-37.91) and 44.4 months (range 0.23-48.62), respectively, while the median BTS was similar, 85.5 mm (IQR 57.2-113.2) and 86.0 mm (IQR 53.0-108.5), respectively (p = 0.42). The median P-group OS was 18.2 months [95% CI 12.2-not reached (NR)] for BTS > 86 mm versus NR (95% CI 27.2-NR) for BTS ≤ 86 mm (p = 0.0026). A high BTS was associated with a shorter OS in univariate analyses (p = 0.009) as well as after adjustment on confounding factors (HR 2.16, [95% CI 1.01-4.65], p = 0.048). The CT-group OS was not statistically different between low and high BTS patients, in univariate and multivariate analyses (p = 0.411).
    CONCLUSIONS: After adjustment on major baseline clinical prognostic factors, BTS was an independent prognostic factor for OS in PD-L1 ≥ 50% advanced NSCLC patients treated first-line with pembrolizumab.
    Keywords:  Baseline tumour size; Immune checkpoint inhibitor; Non-small cell lung cancer; Pembrolizumab
    DOI:  https://doi.org/10.1007/s00262-021-03108-x
  64. Dis Markers. 2021 ;2021 5424623
    MALAT-1 Expression Correlates with Prognosis in Non-Small-Cell Lung Carcinoma: A Systematic Review and Meta-analysis.
    Ran Ran, Jian-Wei Jin, Wei-Ping Zhang.
      Background: Non-small-cell lung carcinoma (abbreviated as NSCLC) progresses rapidly and lacks appropriate biological markers. Recent studies have shown that long noncoding RNA metastasis-associated lung adenocarcinoma transcript 1 (MALAT-1) has potential application value for clinically diagnosing lung carcinoma. Thus, this study conducted a systematic review and meta-analysis for assessing if MALAT-1 has a relationship to NSCLC outcome.Methods: This study conducted the search of China National Knowledge Infrastructure, China Science and Technology Journal, SinoMed, EMBASE, Cochrane library, Web of Science, Wanfang database, and PubMed from inception to September, 1, 2021. The published article about MALAT-l expression for NSCLC patients was analyzed. We used combined hazard rates under the confidence interval of 95% for examining the relationship of MALAT-l and NSCLC.
    Results: In this meta-analysis, we found that 10 studies were included, and MALAT-1 expressions were distinctly related to an unfavorable overall survival (HR: 2.34 (1.65, 3.33); I2 = 76%). Considering the merger's clinical heterogeneity, for meta-analysis, we used the random-effects method.
    Conclusion: Overexpression of MALAT-1 showed correlations to the less effective outcome of NSCLC. MALAT-1 might be a new NSCLC prognosis marker.
    DOI:  https://doi.org/10.1155/2021/5424623
  65. J Basic Clin Physiol Pharmacol. 2021 Dec 03.
    Impact of muscle mass on blood glucose level.
    Murtada Taha, Yaser A AlNaam, Thekra Al Maqati, Lenah Almusallam, Gharam Altalib, Deema Alowfi, Nafish Haider.
      OBJECTIVES: At present, diabetes is one of the leading causes of mortality across the world. It was hypothesized that muscle mass could have a significant influence on blood glucose level and this corelation if established successfully could pave way for novel treatment modalities for type 2 diabetes mellitus (T2DM). In the present study, the association between muscle mass and blood glucose level was examined in a healthy population who was not having T2DM at baseline and was undergoing a regular course of exercise.METHODS: The clinical study was performed involving 53 healthy male populations between 10 and 60 years of age. The participants were sampled in accordance with the quantitative experimental study design, using nonprobability sampling techniques. The independent variable measured among the subjects included muscle mass and blood glucose level, using bioelectrical impedance and a simple glucometer respectively. Subgroup analysis amongst different substantial parameters including body mass index (BMI), myostatin inhibitor usage, and age factor that could affect the muscle mass and glucose level correlation were also studied simultaneously.
    RESULTS: The study findings demonstrated a negative correlation between muscle mass and glucose utilization levels. There was a significant difference in the mean muscle mass of the participants which was 36.2453, and the mean glucose utilization level which was 15.1493%. Pearson correlation between the muscle mass and percentage of glucose utilization of the participants indicated a significant difference (since p-value <0.05) between these two studied parameters.
    CONCLUSIONS: The study finding suggests an inverse association of the skeletal muscle mass with blood glucose level which encourages the implication of muscle-building exercises as the preventive measure for T2DM.
    Keywords:  Pearson correlation; bioelectrical impedance; blood glucose level; glucometer; muscle mass; type 2 diabetes mellitus (T2DM)
    DOI:  https://doi.org/10.1515/jbcpp-2021-0316
  66. Endocr Metab Immune Disord Drug Targets. 2021 Nov 30.
    Progesterone Receptor Membrane Component 1 and Its Accomplice: Emerging Therapeutic Targets in Lung Cancer.
    Solaipriya Solairaja, Satish Ramalingam, Nageswara Rao Dunna, Sivaramakrishnan Venkatabalasubramanian.
      Progesterone receptor membrane component 1 (PGRMC1) is a trans-membrane evolutionarily conserved protein with a cytochrome b5 like heme/steroid binding domain. PGRMC1 clinical levels are strongly suggested to correlate with poor patient survival and lung cancer prognosis. PGRMC1 has been reported to possess pleiotropic functions, such as participating in cellular and membrane trafficking, steroid hormone signaling, cholesterol metabolism and steroidogenesis, glycolysis and mitochondrial energy metabolism, heme transport and homeostasis, neuronal movement and synaptic function, autophagy, anti-apoptosis, stem cell survival and the list is still expanding. PGRMC1 mediates its pleiotropic functions through its ability to interact with multiple binding partners, such as epidermal growth factor receptor (EGFR), sterol regulatory element binding protein cleavage activating protein (SCAP), insulin induced gene-1 protein (Insig-1), heme binding proteins (hepcidin, ferrochelatase and cyp450 members), plasminogen activator inhibitor 1 RNA binding protein (PAIR-BP1). In this review, we provide a comprehensive overview of PGRMC1 and its associated pleiotropic functions that are indispensable for lung cancer promotion and progression, suggesting it as a prospective therapeutic target for intervention. Notably, we have compiled and reported various preclinical studies wherein prospective agonists and antagonists had been tested against PGRMC1 expressing cancer cell lines, suggesting it as a prospective therapeutic target for cancer intervention.
    Keywords:  PGRMC1; binding partners; cancer; chemoresistance; downstream mediators ; heme; tumorigenesis
    DOI:  https://doi.org/10.2174/1871530321666211130145542
  67. Mol Cancer. 2021 12 02. 20(1): 156
    LncRNA PKMYT1AR promotes cancer stem cell maintenance in non-small cell lung cancer via activating Wnt signaling pathway.
    Yaomei He, Xiulin Jiang, Lincan Duan, Qiuxia Xiong, Yixiao Yuan, Peishen Liu, Liping Jiang, Qiushuo Shen, Song Zhao, Cuiping Yang, Yongbin Chen.
      BACKGROUND: Non-small cell lung cancer (NSCLC) is the most common type of human lung cancers, which has diverse pathological features. Although many signaling pathways and therapeutic targets have been defined to play important roles in NSCLC, limiting efficacies have been achieved.METHODS: Bioinformatics methods were used to identify differential long non-coding RNA expression in NSCLC. Real-time RT-PCR experiments were used to examine the expression pattern of lncRNA PKMYT1AR, miR-485-5p. Both in vitro and in vivo functional assays were performed to investigate the functional role of PKMYT1AR/miR-485-5p/PKMYT1 axis on regulating cell proliferation, migration and tumor growth. Dual luciferase reporter assay, fluorescent in situ hybridization (FISH), immunoblot, co-immunoprecipitation experiments were used to verify the molecular mechanism.
    RESULT: Here, we identify a human-specific long non-coding RNA (lncRNA, ENST00000595422), termed PKMYT1AR (PKMYT1 associated lncRNA), that is induced in NSCLC by Yin Yang 1 (YY1) factor, especially in cancerous cell lines (H358, H1975, H1299, H1650, A549 and SPC-A1) compared to that in normal human bronchial epithelium cell line (BEAS-2B). We show that PKMYT1AR high expression correlates with worse clinical outcome, and knockdown of PKMYT1AR inhibits tumor cell proliferation, migration and xenograft tumor formation abilities. Bioinformatic analysis and a luciferase assay demonstrate that PKMYT1AR directly interacts with miR-485-5p to attenuate the inhibitory role on its downstream oncogenic factor PKMYT1 (the protein kinase, membrane-associated tyrosine/threonine 1) in NSCLC. Furthermore, we uncover that miR-485-5p is downregulated in both cancerous cell lines and peripheral blood serum isolated from NSCLC patients compared to reciprocal control groups. Consistently, forced expression of miR-485-5p inhibits the proliferation and migration abilities of tumor cells. Moreover, we provide evidence showing that PKMYT1AR targeting antisense oligonucleotide (ASO) dramatically inhibit tumor growth in vivo. Mechanistic study shows that PKMYT1AR/ miR-485-5p /PKMYT1 axis promotes cancer stem cells (CSCs) maintenance in NSCLC via inhibiting β-TrCP1 mediated ubiquitin degradation of β-catenin proteins, which in turn causes enhanced tumorigenesis.
    CONCLUSIONS: Our findings reveal the critical role of PKMYT1AR/miR-485-5p /PKMYT1 axis during NSCLC progression, which could be used as novel therapeutic targets in the future.
    Keywords:  Cancer stem cells (CSCs); Non-small cell lung cancer; PKMYT1; PKMYT1AR; miR-485-5p
    DOI:  https://doi.org/10.1186/s12943-021-01469-6
  68. Front Cell Dev Biol. 2021 ;9 779367
    Molecular Characterization of the Clinical and Tumor Immune Microenvironment Signature of 5-methylcytosine-Related Regulators in non-small Cell Lung Cancer.
    Taisheng Liu, Liyi Guo, Guihong Liu, Xiaoshan Hu, Xiaoning Li, Jinye Zhang, Zili Dai, Peng Yu, Ming Jiang, Jian Wang, Jian Zhang.
      Background: DNA methylation is an important epigenetic modification, among which 5-methylcytosine methylation (5mC) is generally associated with tumorigenesis. Nonetheless, the potential roles of 5mC regulators in the tumor microenvironment (TME) remain unclear. Methods: The 5mC modification patterns of 1,374 lung adenocarcinoma samples were analyzed systematically. The correlation between the 5mC modification and tumor microenvironment cell infiltration was further assessed. The 5mCscore was developed to evaluate tumor mutation burden, immune check-point inhibitor response, and the clinical prognosis of individual tumors. Results: Three 5mC modification patterns were established based on the clinical characteristics of 21 5mC regulators. According to the differential expression of 5mC regulators, three distinct 5mC gene cluster were also identified, which showed distinct TME immune cell infiltration patterns and clinical prognoses. The 5mCscore was constructed to evaluate the tumor mutation burden, immune check-point inhibitor response, and prognosis characteristics. We found that patients with a low 5mCscore had significant immune cell infiltration and increased clinical benefit. Conclusion: This study indicated that the 5mC modification is involved in regulating TME infiltration remodeling. Targeting 5mC modification regulators might be a novel strategy to treat lung cancer.
    Keywords:  5mC; immunotherapy; lung adenocarcinoma; mutation burden; tumour microenvironment
    DOI:  https://doi.org/10.3389/fcell.2021.779367
  69. Cancer Med. 2021 Dec 04.
    Association of PTPRT mutations with immune checkpoint inhibitors response and outcome in melanoma and non-small cell lung cancer.
    Wenjing Zhang, Fuyan Shi, Yujia Kong, Yuting Li, Chao Sheng, Suzhen Wang, Qinghua Wang.
      PURPOSE: Protein tyrosine phosphatase receptor type T (PTPRT), which is a well-known phosphatase and mutates frequently in melanoma and non-small cell lung cancer (NSCLC). Our research aims to elucidate its mutation association with immune checkpoint inhibitors (ICI) efficacy.METHODS: We integrated whole-exome sequencing (WES)-based somatic mutation profiles and clinical characteristics of 631 melanoma samples received ICI agents from eight studies and 109 NSCLC samples from two studies. For validation, 321 melanoma and 350 NSCLC immunotherapy samples with targeted next-generation sequencing (NGS) were employed. Besides, an independent NSCLC cohort contained 240 samples was also collected for further corroboration. Distinct immune infiltration was evaluated according to the PTPRT mutational status.
    RESULTS: In the WES melanoma cohort, patients with PTPRT mutations harbored a significantly elevated ICI response rate (40.5% vs. 28.6%, p = 0.036) and a prolonged survival outcome (35.3 vs. 24.9 months, p = 0.006). In the WES NSCLC cohort, the favorable response and immunotherapy survival were also observed in PTPRT-mutated patients (p = 0.036 and 0.019, respectively). For the validation cohorts, the associations of PTRPT mutations with better prognoses were identified in melanoma, NSCLC, and pan-cancer patients with targeted-NGS (all p < 0.05). Moreover, immunology analyses showed the higher mutation burden, increased lymphocyte infiltration, decreased- activated-stroma, and immune response pathways were detected in patients with PTPRT mutations.
    CONCLUSION: Our investigation indicates that PTPRT mutations may be considered as a potential indicator for assessing ICI efficacy in melanoma and NSCLC, even across multiple cancers. Further prospective validation cohorts are warranted.
    Keywords:  NSCLC; PTPRT mutation; biomarker; immunotherapy; melanoma
    DOI:  https://doi.org/10.1002/cam4.4472
  70. J Clin Lab Anal. 2021 Dec 03. e24163
    Oxidative stress parameters and keap 1 variants in T2DM: Association with T2DM, diabetic neuropathy, diabetic retinopathy, and obesity.
    Farnaz Khalili, Asad Vaisi-Raygani, Ebrahim Shakiba, Maryam Kohsari, Maryam Dehbani, Rozita Naseri, Soheila Asadi, Ziba Rahimi, Mehrali Rahimi, Zohreh Rahimi.
      INTRODUCTION: Chronic hyperglycemia activates the inflammatory pathways and oxidative stress mechanisms with consequent damage to nerve tissue and retina. The Keap1-Nrf2 pathway acts as one of the most important antioxidant pathways of the organism. Variants of Keap1 could affect susceptibility to diabetes and its complications.METHODS: In a case-control study, 400 individuals included type 2 diabetes mellitus (T2DM) patients without complication, with neuropathy, with retinopathy, and healthy individuals were investigated. The levels of glutathione (GSH), glutathione peroxidase (GPx), malondialdehyde (MDA), and total antioxidant capacity (TAC) were measured using chemical methods. Using the PCR-RFLP method, the Keap1 (rs11085735) variants were identified.
    RESULTS: Neuropathic patients had significantly lower levels of GSH, GPx, and TAC and higher levels of total oxidative status (TOS), MDA, and oxidative stress index (OSI) compared to T2DM patients without complication and controls. Lower levels of GSH and GPx and a higher level of MDA were observed in patients with retinopathy compared with controls. Obesity was associated with significantly lower GPx activity and higher TOS. A significantly higher Keap1 AA genotype was found in patients with neuropathy than T2DM without complication and controls. The presence of Keap1 AA genotype correlated with lower GPx activity compared to CC genotype.
    CONCLUSIONS: Our study suggests the role of reduced antioxidant system and Keap1 variants in the pathogenesis of T2DM and its complications of neuropathy and retinopathy and also obesity in enhanced oxidative stress. Monitoring oxidative stress parameters in diabetic patients, especially those with complication and their treatment with antioxidants is suggested.
    Keywords:  diabetic neuropathy; diabetic retinopathy; keap1 variants; oxidative stress; type 2 diabetes
    DOI:  https://doi.org/10.1002/jcla.24163
  71. Int Urol Nephrol. 2021 Dec 03.
    Intradialytic nutrition and quality of life in Chilean older patients in hemodialysis with protein-energy wasting.
    Mariana Ayala, Margarita Marchant, Cristina Hertz, Gloria Castillo.
      PURPOSE: The study assessed the impact of intradialytic oral nutritional supplementation on the quality of life in patients receiving hemodialysis and diagnosed with protein energy wasting.METHODS: A pre-test post-test quasi-experimental study was conducted before and after 3 months of intradialytic oral nutritional supplementation on 109 older hemodialysis patients. We measured before and after 3 months of intradialytic oral nutritional supplementation, the quality of life score, the burden of kidney disease, three quality of life scales and the mental and physical health status using KDQoL-SF™ 1.3, body composition and biochemical parameters of nutritional condition.
    RESULTS: The mean age of the patients was 69.4 ± 3.4 years, 59% were male, and the time on dialysis was 63.5 ± 52.6 months. Comparing the baseline with month 3 of intradialytic oral nutritional supplementation, we observed to better quality of life. In contrast to malnutrition, score, specifically increased significantly score of symptoms/problems list related to hemodialysis, sexual function, social and cognitive function, sleep, pain, energy/fatigue and general state of health. Significant changes were also found in nutritional status, energy intake and body composition indicators. After 3 months of intradialytic oral nutritional supplementation, we observed a nutritional status recovery in one or more indicators in 92% of the patients.
    CONCLUSION: Our findings indicate that 3 months of intradialysis oral nutritional supplementation improves the components of physical and mental quality of life and nutritional status in older patients receiving hemodialysis diagnosed with loss of protein energy. These results are relevant to improve the experience of patients with protein energy loss receiving hemodialysis.
    Keywords:  Hemodialysis; Intradialytic oral nutritional supplementation; Nutritional status; Protein energy wasting; Quality of life
    DOI:  https://doi.org/10.1007/s11255-021-03077-1
  72. Transl Lung Cancer Res. 2021 Oct;10(10): 3912-3928
    Menstrual factors, reproductive history, and risk of lung cancer: a multi-center population-based cohort study in Chinese females.
    Zhuoyu Yang, Fei Wang, Fengwei Tan, Wei Cao, Yongjie Xu, Chao Qin, Yiwen Yu, Liang Zhao, Yan Wen, Zheng Wu, Yadi Zheng, Yunyong Liu, Lianzheng Yu, Donghua Wei, Dong Dong, Ji Cao, Shaokai Zhang, Shipeng Yan, Ning Wang, Xianzhen Liao, Lingbin Du, Jiang Li, Ni Li, Wanqing Chen, Jie He.
      Background: Female menstrual and reproductive factors, as remarkable indicators of hormone effect, were hypothesized to be associated with lung cancer risk, whereas the existed epidemiological evidence was inconsistent. Our study aims to investigate the association between menstrual and reproductive factors and lung cancer risk based on the Chinese Lung Cancer Screening Program.Methods: This study was based on a large-scale multi-center population cohort across China recruiting individuals aged 40-74 years old between 2013-2018. Cox regression model was applied to estimate the HRs and 95% CIs. Restricted cubic spline (RCS) analysis was used to estimate dose-response relationships and test for nonlinear associations.
    Results: Among 553,434 female participants, 1,529 incident lung cancer cases were identified with a median follow-up of 3.61 years. With adjustment for multiple covariates and all significant hormonal factors, elevated lung cancer risk was associated with later age (15, or ≥16 years) at menarche (HR =1.27, 95% CI: 1.04-1.56; HR =1.45, 95% CI: 1.19-1.76), later age (25-29, or ≥30 years) at first live birth (HR =1.27, 95% CI: 1.13-1.43; HR =1.23, 95% CI: 1.00-1.51), and benign breast disease history (HR =1.25, 95% CI: 1.10-1.41). For postmenopausal females specifically, surgical menopause (HR =1.62; 95% CI: 1.29-2.05) and other surgeries on the reproductive system (HR =1.19; 95% CI: 1.01-1.40) both appeared to be predictive of elevated lung cancer risk. Concerning age at menopause, a nonlinear association was observed (P-nonlinear =0.0126). Increased lung cancer risk was observed among females with age at menopause especially above 50. Although we observed no significant associations between longer time (≥13 months) of breastfeeding and lung cancer risk among all participants (HR =0.86; 95% CI: 0.71-1.04), significant decreased adenocarcinoma risk (HR =0.65; 95% CI: 0.53-0.81) was noted among nonsmoking females.
    Conclusions: Our findings add some support for the role of menstrual and reproductive factors in lung carcinogenesis. However, these relationships were complex, and required further investigations addressing the biological mechanisms.
    Keywords:  Lung cancer; cohort study; hormones; population-based
    DOI:  https://doi.org/10.21037/tlcr-21-552
  73. Eur J Cancer. 2021 Nov 26. pii: S0959-8049(21)01190-4. [Epub ahead of print]
    Failure to EGFR-TKI-based therapy and tumoural progression are promoted by MEOX2/GLI1-mediated epigenetic regulation of EGFR in the human lung cancer.
    Irlanda Peralta-Arrieta, Octavio A Trejo-Villegas, Leonel Armas-López, Hugo A Ceja-Rangel, María Del Carmen Ordóñez-Luna, Priscila Pineda-Villegas, Marco A González-López, Blanca Ortiz-Quintero, Criselda Mendoza-Milla, Zyanya L Zatarain-Barrón, Oscar Arrieta, Joaquín Zúñiga, Federico Ávila-Moreno.
      BACKGROUND: Mesenchyme homeobox-2 (MEOX2)-mediated regulation of glioma-associated oncogene-1 (GLI1) has been associated with poor overall survival, conferring chemoresistance in lung cancer. However, the role of MEOX2/GLI1 in resistance to epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs)-based therapy remains unexplored in human lung cancer.METHODS: Functional assays using genetic silencing strategy by short hairpin RNAs, as well as cytotoxic (tetrazolium dye MTT) and clonogenic assays, were performed to evaluate MEOX2/GLI1-induced malignancy capacity in lung cancer cells. Further analysis performed includes western blot, qPCR and ChIP-qPCR assays to identify whether MEOX2/GLI1 promote EGFR/AKT/ERK activation, as well as EGFR overexpression through epigenetic mechanisms. Finally, preclinical tumour progression in vivo and progression-free disease interval analyses in patients treated with EGFR-TKI were included.
    RESULTS: Overexpressed MEOX2/GLI1 in both EGFR wild-type and EGFR/KRAS-mutated lung cancer cells were detected and involved in the activation/expression of EGFR/AKT/ERK biomarkers. In addition, MEOX2/GLI1 was shown to be involved in the increased proliferation of tumour cells and resistance capacity to cisplatin, EGFR-TKIs (erlotinib and AZD9291 'osimertinib'), AZD8542-SMO, and AZD6244-MEKK1/2. In addition, we identified that MEOX2/GLI1 promote lung tumour cells progression in vivo and are clinically associated with poorer progression-free disease intervals. Finally, both MEOX2 and GLI1 were detected to be epigenetically involved in EGFR expression by reducing both repressive markers polycomb-EZH2 and histone H3K27me3, but, particularly, increasing an activated histone profile H3K27Ac/H3K4me3 at EGFR-gene enhancer-promoter sequences that probably representing a novel EGFR-TKI-based therapy resistance mechanism.
    CONCLUSION: MEOX2/GLI1 promote resistance to cisplatin and EGFR-TKI-based therapy in lung cancer cells, modulating EGFR/AKT/ERK signalling pathway activation, as well as inducing an aberrant epigenetic modulation of the EGFR-gene expression in human lung cancer.
    Keywords:  Cancer drug resistance; Cisplatin; EGFR-TKIs; Epigenetics; Erlotinib; GLI1; Lung cancer; MEOX2; Osimertinib; Transcription factors
    DOI:  https://doi.org/10.1016/j.ejca.2021.10.032
  74. ACS Omega. 2021 Nov 23. 6(46): 31272-31281
    WSF-CT-11, a Sesquiterpene Derivative, Activates AMP-Activated Protein Kinase with Anti-diabetic Effects in 3T3-L1 Adipocytes.
    Yang Yang, Yunyun Wang, Zhijie Zhang, Shifa Wang, Zhen Li.
      BACKGROUND: AMP-activated protein kinase (AMPK) is a therapeutic target against type II diabetes (T2D). Synthetic sesquiterpene derivatives were investigated to identify novel AMPK activators as anti-diabetic drugs because the leading drugs like metformin and thiazolidinediones (TZDs) activate AMPK by inhibiting the synthesis of adenosine 5'-triphosphate and thus are associated with some side effects.RESULTS: We identified WSF-CT-11 as an AMPK activator in HEK293T cells and found that WSF-CT-11 activates AMPK by the activation of transient receptor potential vanilloid type 1 (TRPV1), a Ca2+-permeable cation channel. The increased Ca2+ influx then activates phosphoinositide 3-kinase (PI3K), protein kinase B (PKB/Akt), and Ca2+/calmodulin-dependent protein kinase II (CaMKII), which in turn phosphorylates TRPV1 and facilitates the formation of a TRPV1/Akt/CaMKII/AMPK complex. This complex might be important for the regulation of AMPK activity. In 3T3-L1 adipocytes, WSF-CT-11-induced AMPK activation has three anti-diabetic effects. It promotes the assembly of high-molecular-weight adiponectin, which has stronger insulin-sensitizing activity than other multimers. It improves translocation of the glucose transporter type 4, increases glucose uptake, and induces the inhibitory phosphorylation of peroxisome proliferator-activated receptor γ and thus suppresses adipogenesis.
    CONCLUSION: WSF-CT-11 is a novel AMPK activator and potential drug against T2D without the side effects of metformin and TZDs.
    DOI:  https://doi.org/10.1021/acsomega.1c05061
  75. Clin Nutr ESPEN. 2021 Dec;pii: S2405-4577(21)01047-0. [Epub ahead of print]46 87-98
    Comparison of prophylactic percutaneous endoscopic gastrostomy with reactive enteral nutrition in patients with head and neck cancer undergoing radiotherapy or chemoradiotherapy: A systematic review.
    Kate Mellors, Xiaodan Ye, Jonathan Van Den Brande, Tsz Wai Ray Mak, Teresa Brown, Merran Findlay, Judy Bauer.
      BACKGROUND & AIMS: Nutrition support is frequently indicated in patients with head and neck cancer (HNC). However, the optimal timing of enteral tube placement and feeding commencement is unknown. This review aims to compare the outcomes for patients with HNC undergoing curative intent radiotherapy (RT) or chemoradiotherapy (CRT) receiving either prophylactic percutaneous endoscopic gastrostomy (pPEG) tube placement/feeding or reactive enteral nutrition (rEN).METHODS: A literature search was conducted in March 2020 across PubMed, CINAHL, Embase, Web of Science, and Scopus. Randomized controlled trials (RCTs) of patients (≥18 years) with HNC who had received either pPEG or rEN were included. Outcomes examined were weight change, nutritional status, body mass index, treatment interruptions, quality of life (QoL), disease-free survival and overall survival. Study quality and certainty of evidence were assessed using the Cochrane Risk-of-bias Tool for Randomized Trials Version 2 and the Grading of Recommendations Assessment, Development and Evaluation system, respectively.
    RESULTS: Five studies (three RCTs) (n = 298) were included and definitions of pPEG and rEN were heterogenous. pPEG was associated with a clinically important reduction in short-term critical weight loss (>10% weight loss), and significantly improved short-term QoL in patients with HNC. The timing of nutrition support commencement had no effect on all other outcomes. The overall certainty of evidence was 'moderate' for: nutritional status; treatment interruptions; short-term QoL; disease-free survival; and 'low' for all other outcomes.
    CONCLUSIONS: Patients with HNC undergoing RT or CRT receiving pPEG tube feeding/placement were less likely to experience short-term critical weight loss and have improved short-term QoL compared to rEN. Further well-designed RCTs with consistent definitions of tube feeding protocols and the use of validated tools to evaluate nutritional status, will assist to increase the certainty of evidence and confirm the beneficial effects observed.
    Keywords:  Head and neck cancer; Nasogastric tube; Nutritional status; Prophylactic gastrostomy; Weight loss
    DOI:  https://doi.org/10.1016/j.clnesp.2021.09.724
  76. Nat Rev Drug Discov. 2021 Dec 03.
    Targeting cancer metabolism in the era of precision oncology.
    Zachary E Stine, Zachary T Schug, Joseph M Salvino, Chi V Dang.
      One hundred years have passed since Warburg discovered alterations in cancer metabolism, more than 70 years since Sidney Farber introduced anti-folates that transformed the treatment of childhood leukaemia, and 20 years since metabolism was linked to oncogenes. However, progress in targeting cancer metabolism therapeutically in the past decade has been limited. Only a few metabolism-based drugs for cancer have been successfully developed, some of which are in - or en route to - clinical trials. Strategies for targeting the intrinsic metabolism of cancer cells often did not account for the metabolism of non-cancer stromal and immune cells, which have pivotal roles in tumour progression and maintenance. By considering immune cell metabolism and the clinical manifestations of inborn errors of metabolism, it may be possible to isolate undesirable off-tumour, on-target effects of metabolic drugs during their development. Hence, the conceptual framework for drug design must consider the metabolic vulnerabilities of non-cancer cells in the tumour immune microenvironment, as well as those of cancer cells. In this Review, we cover the recent developments, notable milestones and setbacks in targeting cancer metabolism, and discuss the way forward for the field.
    DOI:  https://doi.org/10.1038/s41573-021-00339-6
  77. J Cachexia Sarcopenia Muscle. 2021 Dec 04.
    Development and validation of a Modified Patient-Generated Subjective Global Assessment as a nutritional assessment tool in cancer patients.
    INSCOC Study Group
      BACKGROUND: Completing Patient-Generated Subjective Global Assessment (PG-SGA) questionnaires is time consuming. This study aimed to develop and validate an easy-to-use modified PG-SGA (mPG-SGA) for cancer patients.METHODS: Seventy professionals assessed the content validity, comprehensibility, and difficulty of the full PG-SGA. A survey including the PG-SGA and other questionnaires was completed by 34 071 adult hospitalized cancer patients with first cancer diagnosis or recurrent disease with any tumour comorbidities from the INSCOC study. Among them, 1558 patients were followed for 5 years after admission. Reliability and rank correlation were estimated to assess the consistency between PG-SGA items and to select mPG-SGA items. The external and internal validity, test-retest reliability, and predictive validity were tested for the mPG-SGA via comparison with both the PG-SGA and abridged PG-SGA (abPG-SGA).
    RESULTS: After deleting items that more than 50% of professionals considered difficult to evaluate (Worksheet 4) and items with an item-total correlation <0.1, the mPG-SGA was constructed. Nutritional status was categorized using mPG-SGA scores as well-nourished (0 points) or mildly (1-2 points), moderately (3-6 points), or severely malnourished (≥7 points) based on the area under curve (0.962, 0.989, and 0.985) and maximal sensitivity (0.924, 0.918, and 0.945) and specificity (1.000, 1.000, and 0.938) of the cut-off scores. The external and internal validity and test-retest reliability were good. Significant median overall survival differences were found among nutritional status groups categorized by the mPG-SGA: 24, 18, 14, and 10 months for well-nourished, mildly malnourished, moderately malnourished, and severely malnourished, respectively (all Ps < 0.05). Neither the PG-SGA nor the abridged PG-SGA could discriminate the median overall survival differences between the well-nourished and mildly malnourished groups.
    CONCLUSIONS: We systematically developed and validated the mPG-SGA as an easier-to-use nutritional assessment tool for cancer patients. The mPG-SGA appears to have better predictive validity for survival than the PG-SGA and abridged PG-SGA.
    Keywords:  Cancer patient; Nutritional assessment tool; PG-SGA; Patient-generated subjective global assessment; mPG-SGA
    DOI:  https://doi.org/10.1002/jcsm.12872
  78. Aging (Albany NY). 2021 Dec 01. 13(undefined):
    Construction and validation of a metabolic gene-associated prognostic model for cervical carcinoma and the role on tumor microenvironment and immunity.
    Jinzhi Huang, Fei Luo, Mingjie Shi, Jiaxin Luo, Choudi Ma, Shangzheng Li, Yue Wei, Runmin Guo, Ting Li.
      Metabolic reprogramming is a common feature of tumor cells and is associated with tumorigenesis and progression. In this study, a metabolic gene-associated prognostic model (MGPM) was constructed using multiple bioinformatics analysis methods in cervical carcinoma (CC) tissues from The Cancer Genome Atlas (TCGA) database, which comprised fifteen differentially expressed metabolic genes (DEMGs). Patients were divided into a high-risk group with shorter overall survival (OS) and a low-risk group with better survival. Receiver operating characteristic (ROC) curve analysis showed that the MGPM precisely predicted the 1-, 3- and 5-year survival of CC patients. As expected, MGPM exhibited a favorable prognostic significance in the training and testing datasets of TCGA. And the clinicopathological parameters including stage, tumor (T) and metastasis (M) classifications had significant differences in low- and high-risk groups, which further demonstrated the MGPM had a favorite prognostic prediction ability. Additionally, patients with low-ESTMATEScore had a shorter OS and when those combined with high-risk scores presented a worse prognosis. Through "CIBERSORT" package and Wilcoxon rank-sum test, patients in the high-risk group with a poor prognosis showed lower levels of infiltration of T cell CD8 (P < 0.001), T cells memory activated (P = 0.010) and mast cells resting (P < 0.001), and higher levels of mast cells activated (P < 0.001), and we also found these patients had a worse response for immunosuppressive therapy. These findings demonstrate that MGPM accurately predicts survival outcomes in CC patients, which will be helpful for further optimizing immunotherapies for cancer by reprogramming its cell metabolism.
    Keywords:  cervical carcinoma; immunity; metabolism; prognosis; tumor microenvironment
    DOI:  https://doi.org/10.18632/aging.203723
  79. Biochim Biophys Acta Mol Cell Biol Lipids. 2021 Nov 29. pii: S1388-1981(21)00209-2. [Epub ahead of print] 159081
    Ghrelin is a signal to facilitate the utilization of fatty acids and save glucose by the liver, skeletal muscle, and adipose tissues in chicks.
    Xixi Song, Minghui Wang, Hongchao Jiao, Jingpeng Zhao, Xiaojuan Wang, Hai Lin.
      Ghrelin, classically known as a central appetite-stimulating hormone, has recently been recognized to play an important role in peripheral tissue energy metabolism. In chicken, contrary to mammal, ghrelin acts as an anorexia signal, increased by fasting and further elevated after refed. In the present study, the effect of ghrelin on glucose/lipid utilization by peripheral tissues was investigated. Injection of exogenous acyl ghrelin reduced plasma triglyceride and glucose levels of chickens at both fasting and fed status. In the in vitro cultured chicken primary hepatocytes, adipocytes, and myoblasts, ghrelin suppressed glucose uptake, stimulated fatty acids uptake and oxidation, and decreased TG content. In hepatocyte, ghrelin increased the activities of LPL and HL, and upregulated the expression levels of gene ACC, CPT1, and PPARα. Ghrelin treatment markedly increased the protein level of p-ACC, PPARγ, PGC1α, and CPT1 in hepatocytes, adipocytes and myoblasts. Inhibition of AMPK activity by Compound C had no influence on glucose uptake by hepatocyte, adipocyte, and myoblast, but further amplified the stimulated fatty acid uptake of adipocyte by ghrelin. The present result demonstrates that ghrelin facilitates the uptake and oxidation of fatty acid and cut down the utilization of glucose by the liver, muscle, and adipose tissues. The result suggests that ghrelin functions as a signal of fatty acid oxidation. The study provides a vital framework for understanding the intrinsic role of ghrelin as a crucial factor in the concerted regulation of metabolic substrate of hepatocytes, adipocytes, and myoblasts.
    Keywords:  AMPK; Chick; Fat metabolism; Fatty acids; Ghrelin; Glucose
    DOI:  https://doi.org/10.1016/j.bbalip.2021.159081
  80. Physiol Biochem Zool. 2022 Jan-Feb;95(1):95(1): 35-53
    Altered Oxidative Status as a Cost of Reproduction in a Seabird with High Reproductive Costs.
    Yimei Lin, Allison Patterson, Ana Gabriela Jimenez, Kyle Elliott.
      AbstractLife history theory posits that reproduction is constrained by a cost of reproduction such that any increase in breeding effort should reduce subsequent survival. Oxidative stress refers to an imbalance between the prooxidant reactive oxygen species (ROS) and antioxidant defense. If not thwarted, ROS can cause damage to DNA, lipids, and proteins, potentially increasing the rate of senescence and decreasing cellular function. Reproduction is often associated with higher metabolic rates, which could increase production of ROS and lead to oxidative damage if the animal does not increase antioxidant protection. Thus, oxidative stress could be one mechanism creating a cost of reproduction. In this study we explored how reproduction may affect oxidative status differently between male and female thick-billed murres during early and late breeding seasons over three consecutive years. We manipulated breeding efforts by removing an egg from the nest of some individuals, which forced females to relay, and by handicapping other individuals by clipping wings. We measured total antioxidant capacity (TAC), uric acid (UA) concentration, and malondialdehyde (MDA; an index of lipid oxidative damage) concentration in blood plasma as well as activities of superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT) in red blood cells. Oxidative status was highly variable across years, and year was consistently the most important factor determining oxidative status; inconsistent results in previous field studies may be because reproductive oxidative stress occurs only in some years. Females had lower SOD and GPx and higher MDA and TAC than males immediately after egg laying, suggesting that the cost of egg laying required investment in cheaper nonenzymatic antioxidant defenses that had lower capacity for defending against lipid peroxidation. Delayed birds had lower UA and lower SOD, GPx, and CAT activity compared with control birds. In conclusion, when reproductive costs increase via higher energy costs or longer breeding seasons, the oxidative status of both male and female murres deteriorated as a result of reduced antioxidant defenses.
    Keywords:  antioxidants; life history strategies; offspring care; oxidative stress; reproduction; thick-billed murre
    DOI:  https://doi.org/10.1086/717916
  81. Am J Respir Crit Care Med. 2021 Dec 01. 204(11): P19-P20
    Lung Cancer Screening.
    Yaron B Gesthalter, Eric J Seeley.
      
    DOI:  https://doi.org/10.1164/rccm.20411P19
  82. Mol Neurobiol. 2021 Dec 02.
    Metabolic Profile in Plasma AND CSF of LEVODOPA-induced Dyskinesia in Parkinson's Disease: Focus on Neuroinflammation.
    Bruno L Santos-Lobato, Luiz Gustavo Gardinassi, Mariza Bortolanza, Ana Paula Ferranti Peti, Ângela V Pimentel, Lúcia Helena Faccioli, Elaine A Del-Bel, Vitor Tumas.
      The existence of few biomarkers and the lack of a better understanding of the pathophysiology of levodopa-induced dyskinesia (LID) in Parkinson's disease (PD) require new approaches, as the metabolomic analysis, for discoveries. We aimed to identify a metabolic profile associated with LID in patients with PD in an original cohort and to confirm the results in an external cohort (BioFIND). In the original cohort, plasma and CSF were collected from 20 healthy controls, 23 patients with PD without LID, and 24 patients with PD with LID. LC-MS/MS and metabolomics data analysis were used to perform untargeted metabolomics. Untargeted metabolomics data from the BioFIND cohort were analyzed. We identified a metabolic profile associated with LID in PD, composed of multiple metabolic pathways. In particular, the dysregulation of the glycosphingolipid metabolic pathway was more related to LID and was strongly associated with the severity of dyskinetic movements. Furthermore, bile acid biosynthesis metabolites simultaneously found in plasma and CSF have distinguished patients with LID from other participants. Data from the BioFIND cohort confirmed dysregulation in plasma metabolites from the bile acid biosynthesis pathway. There is a distinct metabolic profile associated with LID in PD, both in plasma and CSF, which may be associated with the dysregulation of lipid metabolism and neuroinflammation.
    Keywords:  Dyskinesia; Levodopa; Metabolomics; Parkinson’s disease
    DOI:  https://doi.org/10.1007/s12035-021-02625-1
  83. Cell Metab. 2021 Nov 24. pii: S1550-4131(21)00531-3. [Epub ahead of print]
    Cysteine 253 of UCP1 regulates energy expenditure and sex-dependent adipose tissue inflammation.
    Evanna L Mills, Cathal Harmon, Mark P Jedrychowski, Haopeng Xiao, Anja V Gruszczyk, Gary A Bradshaw, Nhien Tran, Ryan Garrity, Dina Laznik-Bogoslavski, John Szpyt, Hannah Prendeville, Lydia Lynch, Michael P Murphy, Steven P Gygi, Bruce M Spiegelman, Edward T Chouchani.
      Uncoupling protein 1 (UCP1) is a major regulator of brown and beige adipocyte energy expenditure and metabolic homeostasis. However, the widely employed UCP1 loss-of-function model has recently been shown to have a severe deficiency in the entire electron transport chain of thermogenic fat. As such, the role of UCP1 in metabolic regulation in vivo remains unclear. We recently identified cysteine-253 as a regulatory site on UCP1 that elevates protein activity upon covalent modification. Here, we examine the physiological importance of this site through the generation of a UCP1 cysteine-253-null (UCP1 C253A) mouse, a precise genetic model for selective disruption of UCP1 in vivo. UCP1 C253A mice exhibit significantly compromised thermogenic responses in both males and females but display no measurable effect on fat accumulation in an obesogenic environment. Unexpectedly, we find that a lack of C253 results in adipose tissue redox stress, which drives substantial immune cell infiltration and systemic inflammatory pathology in adipose tissues and liver of male, but not female, mice. Elevation of systemic estrogen reverses this male-specific pathology, providing a basis for protection from inflammation due to loss of UCP1 C253 in females. Together, our results establish the UCP1 C253 activation site as a regulator of acute thermogenesis and sex-dependent tissue inflammation.
    Keywords:  UCP1; cysteine; inflammation; metabolism; reactive oxygen species; sex differences
    DOI:  https://doi.org/10.1016/j.cmet.2021.11.003
  84. Exp Clin Endocrinol Diabetes. 2021 Dec 02.
    Comparison of Beinaglutide Versus Metformin for Weight Loss in Overweight and Obese Non-diabetic Patients.
    Lijun Gao, Hong Huang, Lu Zhang, Ningjing Zhang, Yuzhe Fu, Dalong Zhu, Yan Bi, Wenhuan Feng.
      PURPOSE: We compared the efficacy and safety of beinaglutide, a glucagon-like peptide-1 (GLP-1) analogue with metformin in lowering the bodyweight of patients who were overweight/obese and non-diabetic.PATIENTS AND METHODS: Seventy-eight non-diabetic patients were randomly selected and beinaglutide or metformin was administered for 12 weeks. The primary endpoints were changes in body weight and the proportions of patients who lost≥5 and≥10% of their baseline body weights.
    RESULTS: A total of 64 patients completed the study; patients in the beinaglutide group exhibited more bodyweight loss than those in the metformin group [(9.5±0.8%; 9.1±0.9 kg) and (5.1±0.9%; 4.5±0.8 kg), respectively, corresponding to a difference of approximately 4.5 kg (95% confidence interval, 2.2-6.9 kg; P<0.01)]. In the beinaglutide group, 90.6 and 40.6% of the patients lost≥5 and≥10% of their body weight, respectively, whereas, in the metformin group, these rates were 46.9 and 12.5%, respectively (P<0.01 and P<0.05). Weight loss following beinaglutide treatment mainly resulted from the loss of fat mass. Compared to metformin, beinaglutide induced a greater decrease in the body mass index, weight circumference, percent body fat, and body fat mass (total, trunk, limb, android, and gynoid). Additionally, beinaglutide decreased serum insulin levels and ameliorated insulin resistance.
    CONCLUSIONS: Beinaglutide is more efficient than metformin at reducing weight and fat mass in patients who are overweight/obese and non-diabetic. Beinaglutide may be a useful therapeutic option for overweight/obesity control in the Chinese population.
    DOI:  https://doi.org/10.1055/a-1608-0345
  85. J Food Biochem. 2021 Dec 01. e14002
    Anti-obesity effect of Auricularia heimuer fruiting body alcohol extraction on obese mice and crucial metabolite pathway analysis by liquid chromatography-tandem mass spectrometry.
    Juan Yang, Tianfeng Zhou, Linxiang Huang, Bingzhi Chen, Yuji Jiang.
      In recent years, the increasing obese and overweight population has become a worldwide public health problem, as there is no effective medication to control obesity. Auricularia heimuer is rich in active substances that have potential biologically active functions. The anti-obesity effect and mechanism of Auricularia heimuer fruiting body alcohol extraction (AHA, 150-600 mg/kg·bw) was investigated in obese mice by assessing changes in endogenous liver metabolites using a liquid chromatography-tandem mass spectrometry approach. The aim of this study was to identify an effective food to control human obesity. AHA of 600 mg/kg·bw (HC) significantly decreased body weight and improved serum biochemistry indices. Sixty-eight liver metabolites were identified and significantly separated among the normal, high-fat diet (HFD), and HC groups. Moreover, the metabolic analysis revealed that HC significantly regulated specific metabolites in mice including amino acids, lipids, and carbohydrate compounds. Kyoto Encyclopedia of Genes and Genomes enrichment analysis revealed that HC was significantly involved in different metabolite pathways including arachidonic acid metabolism, galactose metabolism, carbohydrate digestion and absorption, linoleic acid metabolism, and starch and sucrose metabolism. Eight weeks after supplementing with HC, major metabolites in related pathways that were disrupted by an HFD were restored to normal levels, suggesting that HC had anti-obesity activity.
    Keywords:   Auricularia heimuer ; alcohol extraction; high-fat diet; metabolites analysis
    DOI:  https://doi.org/10.1111/jfbc.14002
  86. Oncology. 2021 Nov 29.
    Post-Progression Survival Highly Influences Overall Survival in Driver Gene Mutation/Translocation Negative or Unknown Type of Non-Small Cell Lung Cancer.
    Hisao Imai, Ryoichi Onozato, Kyoichi Kaira, Sayaka Kawashima, Ken Masubuchi, Kohei Tajima, Koichi Minato.
      INTRODUCTION: In stage I-III non-small cell lung cancer (NSCLC), which is considered operable, surgical resection is the most efficacious treatment and is considered to provide a cure. However, after complete surgical resection, approximately 50% of patients with stage I-IIIA NSCLC experience recurrence and death. Once postoperative recurrence of NSCLC occurs, the prognosis is significantly poor, and the course of treatment after recurrence may influence overall survival (OS). Consequently, we investigated the relationship between relapse-free survival (RFS), post-progression survival (PPS), and OS in patients with postoperative recurrence of NSCLC with driver gene mutation/translocation negative or unknown status.METHODS: Between January 2007 and September 2019, 101 patients with driver gene mutation/translocation negative or unknown status of NSCLC who underwent complete resection and in whom recurrence occurred were analyzed. The associations between RFS, PPS, and OS were analyzed at the individual patient level.
    RESULTS: Linear regression and Spearman rank correlation analyses revealed that PPS was strongly associated with OS (r = 0.83, p < 0.0001, R2 = 0.71), whereas RFS was moderately correlated with OS (r = 0.65, p < 0.0001, R2 = 0.48). In the multivariate analysis, performance status at relapse, administration of immune checkpoint inhibitors, and radiotherapy for oligo-recurrences were significantly associated with PPS (p < 0.001).
    CONCLUSION: Current analysis of individual-level data of patients who underwent complete resection implied that PPS had a higher impact on OS than RFS in patients with postoperative recurrence of driver gene mutation/translocation negative or unknown status of NSCLC. Additionally, current perceptions indicate that treatment beyond progression after complete surgical resection might strongly affect OS.
    DOI:  https://doi.org/10.1159/000521141
  87. Transl Lung Cancer Res. 2021 Oct;10(10): 3902-3911
    Are there differences by sex in lung cancer characteristics at diagnosis? -a nationwide study.
    Alberto Ruano-Ravina, Mariano Provencio, Virginia Calvo de Juan, Enric Carcereny, Anna Estival, Delvys Rodríguez-Abreu, Gretel Benítez, Rafael López-Castro, Marta Belver, María Guirado-Risueño, Carlos Guirao-Rubio, Ana Blasco, Bartomeu Massutí, Ana Laura Ortega, Manuel Cobo, Joaquín Mosquera-Martínez, Carlos Aguado de la Rosa, Joaquim Bosch-Barrera, Amparo Sánchez-Gastaldo, Edel Del Barco Morillo, Óscar Juan, Manuel Dómine, José Manuel Trigo, Diego Pereiro Corbacho, Juana Oramas.
      Background: Lung cancer causes approximately 25% of all cancer deaths. Despite its relevance, few studies have analyzed differences by sex at the time of diagnosis in terms of symptoms, stage, age or smoking status. We aim to assess if there are differences between men and women on these characteristics at diagnosis.Methods: We analyzed the Thoracic Tumour Registry (TTR), sponsored by the Spanish Lung Cancer Group using a case-series design. This is a nationwide registry of lung cancer cases which started recruitment in 2016. For each case included, clinicians fulfilled an electronic record registering demographic data, symptoms, exposure to lung cancer risk factors, and treatment received in detail. We compared men and women using descriptive statistics.
    Results: A total of 13,590 participants took part in this study, 25.6% women. Women were 4 years younger than men (64 vs. 69), and men had smoked more frequently. Adenocarcinoma was the most frequent histological type in both sexes. Stage IV at diagnosis was 50.8% in women compared to 43.6% in men. Weight loss/anorexia/asthenia was the most frequent symptom in both sexes and there were no differences in the number of symptoms at diagnosis. There were no relevant differences in the frequency or number of symptoms by sex when non-small cell lung cancer (NSCLC) and small-cell lung cancer (SCLC) were analyzed separately. Smoking status did not appear to cause different lung cancer presentation in men compared to women.
    Conclusions: There seems to be no differences in lung cancer characteristics by sex at the time at diagnosis on stage, specific symptoms or number of symptoms.
    Keywords:  Lung neoplasms; sex; small cell lung cancer (SCLC); smoking; symptoms
    DOI:  https://doi.org/10.21037/tlcr-21-559
  88. Clin Transl Med. 2021 11;11(11): e588
    Therapeutic targeting of SPIB/SPI1-facilitated interplay of cancer cells and neutrophils inhibits aerobic glycolysis and cancer progression.
    Jianqun Wang, Xiaojing Wang, Yanhua Guo, Lin Ye, Dan Li, Anpei Hu, Shuang Cai, Boling Yuan, Shikai Jin, Yi Zhou, Qilan Li, Liduan Zheng, Qiangsong Tong.
      BACKGROUND: As a metabolic reprogramming feature, cancer cells derive most of their energy from aerobic glycolysis, while its regulatory mechanisms and therapeutic strategies continue to be illusive.METHODS: Integrative analysis of publically available expression profile datasets was used to identify critical transcriptional regulators and their target glycolytic enzymes. The functions and acting mechanisms of transcriptional regulators in cancer cells were investigated by using in vitro and in vivo assays. The Kaplan-Meier curve and log-rank assay were used to conduct the survival study.
    RESULTS: Salmonella pathogenicity island 1 (SPI1/PU.1), a haematopoietic transcription factor, was identified to facilitate glycolytic process, tumourigenesis, invasiveness, as well as metastasis of colon cancer cells, which was interplayed by tumour-associated neutrophils. Mechanistically, neutrophils delivered SPI1 mRNA via extracellular vesicles, resulting in enhanced SPI1 expression of cancer cells. Through physical interaction with SPI1-related protein (SPIB), SPI1 drove expression of glycolytic genes within cancer cells, which in turn induced polarization of neutrophils via glycolytic metabolite lactate. Depletion of neutrophils or SPIB-SPI1 interaction in cancer cells significantly inhibited glycolytic process, tumourigenesis and aggressiveness. Upregulation of SPI1 or SPIB was found to be associated with poor prognosis in patients suffering from colon cancer.
    CONCLUSIONS: Therapeutic targeting of SPIB/SPI1-facilitated interplay of cancerous cells and neutrophils suppresses aerobic glycolysis and progression of cancer.
    Keywords:  SPI1-related protein; Salmonella pathogenicity island 1; aerobic glycolysis; cancer progression; extracellular vesicles; neutrophil
    DOI:  https://doi.org/10.1002/ctm2.588
  89. Sci Rep. 2021 Dec 02. 11(1): 23337
    Analysis at the single-cell level indicates an important role of heterogeneous global DNA methylation status on the progression of lung adenocarcinoma.
    Quan-Fang Chen, Han Gao, Qing-Yun Pan, Ying-Ju Wang, Xiao-Ning Zhong.
      Aberrant DNA modifications affect the tumorigenesis and progression of lung cancer. However, the global methylation status of tumor cells and the heterogeneous methylation status of cells within the same tumor need further study. We used publicly available single-cell RNAseq data to investigate the impact and diversity of global methylation status on lung adenocarcinoma. Clustering cells into subgroups and cell differentiation pseudotime analysis, based on expression profile, demonstrated that the global methylation status was crucial to lung adenocarcinoma function and progression. Hypermethylated tumor cells had increased activity related to the hypoxia response. Hyper- and hypomethylated cells indicated upregulation in pathways involving focal adhesion and cell junctions. Pseudotime analysis showed that cell clusters with unique methylation activities were located at the ends of the putative trajectories, suggesting that DNA methylation and demethylation activities were essential to tumor cell progression. Expression of SPP1 was associated with the global methylation status of tumor cells and with patient prognosis. Our study identified the importance and diversity of global DNA methylation status by analysis at the single-cell level. Our findings provide new information about the global DNA methylation status of tumor cells and suggest new approaches for precision medical treatments for lung adenocarcinoma.
    DOI:  https://doi.org/10.1038/s41598-021-02786-y
  90. Clin Lung Cancer. 2021 Oct 21. pii: S1525-7304(21)00245-X. [Epub ahead of print]
    Impact of Tumor Suppressor Gene Co-Mutations on Differential Response to EGFR TKI Therapy in EGFR L858R and Exon 19 Deletion Lung Cancer.
    Jessica A Hellyer, Maya N White, Rebecca M Gardner, Kristen Cunanan, Sukhmani K Padda, Millie Das, Kavitha Ramchandran, Joel W Neal, Heather A Wakelee.
      BACKGROUND: In most studies, patients with EGFR L858R mutant non-small cell lung cancer (NSCLC) have a shorter duration of response to EGFR tyrosine kinase inhibitor (TKI) therapy than do patients with EGFR exon 19 deletion NSCLC. The role that co-mutations play in this observation is unknown.METHODS: We performed a single-institution retrospective analysis of patients with EGFR-mutant NSCLC (exon 19 deletion or L858R mutation) who received frontline EGFR TKI for metastatic disease between 2014 and 2019, and who had STAMP next-generation sequencing (NGS), a 130-gene platform. Time to treatment failure (TTF) and overall survival were calculated using Cox models adjusted for age, race, and brain metastases. Co-mutations in key tumor suppressor genes (TP53, RB1, KEAP1, CDKN2A, or CTNNB1) were identified and their effects on outcomes were evaluated. Analyses were stratified according to receipt of osimertinib versus nonosimertinib as frontline EGFR TKI.
    RESULTS: Of 137 patients, 72 (57%) had EGFR exon 19 deletions and 65 (43%) had EGFR L858R mutations. Median TTF and OS on frontline TKI was shorter for the L858R cohort versus the exon 19 deletion cohort in univariate analysis. In adjusted models, this difference persisted for TTF but was no longer significant for OS. The difference in TTF in L858R mutant tumors was driven by the presence of co-mutations in key tumor suppressor genes.
    CONCLUSION: Patients with metastatic NSCLC with mutations in EGFR L858R had shorter TTF on frontline TKI compared to patients with EGFR exon 19 deletions. Co-mutations in tumor suppressor genes may play an important role in the differential response to TKI therapy.
    Keywords:  Driver mutations; Gene interaction; NSCLC; Osimertinib; Targeted therapy
    DOI:  https://doi.org/10.1016/j.cllc.2021.09.004
  91. Lung Cancer. 2021 Oct 19. pii: S0169-5002(21)00578-X. [Epub ahead of print]
    Clinical validation of Guardant360 CDx as a blood-based companion diagnostic for sotorasib.
    Joshua M Bauml, Bob T Li, Vamsidhar Velcheti, Ramaswamy Govindan, Alessandra Curioni-Fontecedro, Christophe Dooms, Toshiaki Takahashi, Andrew W Duda, Justin I Odegaard, Fernando Cruz-Guilloty, Liming Jin, Ying Zhang, Abraham Anderson, Ferdinandos Skoulidis.
      OBJECTIVES: Effective therapy for non-small-cell lung cancer (NSCLC) depends on morphological and genomic classification, with comprehensive screening for guideline-recommended biomarkers critical to guide treatment. Companion diagnostics, which provide robust genotyping results, represent an important component of personalized oncology. We evaluated the clinical validity of Guardant360 CDx as a companion diagnostic for sotorasib for detection of KRAS p.G12C, an important oncogenic NSCLC driver mutation.MATERIALS AND METHODS: KRAS p.G12C was tested in NSCLC patients from CodeBreaK100 (NCT03600833) in pretreatment plasma samples using Guardant360 CDx liquid biopsy and archival tissue samples using therascreen® KRAS RGQ polymerase chain reaction (PCR) kit tissue testing. Matched tissue and plasma samples were procured from other clinical trials or commercial vendors, and results were compared. Demographics and clinical characteristics and objective response rate (ORR) were evaluated.
    RESULTS: Of 126 CodeBreaK patients, 112 (88.9%) were tested for KRASp.G12C mutations with Guardant360 CDx. Among 189 patients in the extended analysis cohort, the positive and negative percent agreement (95% CI) for Guardant360 CDx plasma testing relative to therascreen® KRAS RGQ PCR kit tissue testing were 0.71 (0.62, 0.79) and 1.00 (0.95, 1.00), respectively; overall percent agreement (95% CI) was 0.82 (0.76, 0.87). TP53 co-mutations were the most common regardless of KRAS p.G12C status (KRAS p.G12C-positive, 53.4%; KRAS p.G12C-negative, 45.5%). STK11 was co-mutated in 26.1% of KRAS p.G12C-positive samples. The ORR was similar among patients selected by plasma and tissue testing.
    CONCLUSION: Comprehensive genotyping for all therapeutic targets including KRAS p.G12C is critical for management of NSCLC. Liquid biopsy using Guardant360 CDx has clinical validity for identification of patients with KRASp.G12C-mutant NSCLC and, augmented by tissue testing methodologies as outlined on the approved product label, will identify patients for treatment with sotorasib.
    Keywords:  Biomarkers; Carcinoma, non–small-cell lung; Liquid biopsy; Molecular diagnostic techniques; Sotorasib; Tumor
    DOI:  https://doi.org/10.1016/j.lungcan.2021.10.007
  92. Nanomedicine (Lond). 2021 Dec 02.
    Immunotherapy for cancer: effects of iron oxide nanoparticles on polarization of tumor-associated macrophages.
    Camila Sales Nascimento, Érica Alessandra Rocha Alves, Celso Pinto de Melo, Rodrigo Corrêa-Oliveira, Carlos Eduardo Calzavara-Silva.
      Cancer immunotherapy is the most promising trend in oncology, focusing on helping or activating the patient's immune system to identify and fight against cancer. In the last decade, interest in metabolic reprogramming of tumor-associated macrophages from M2-like phenotype (promoting tumor progression) to M1-like phenotypes (suppressing tumor growth) as a therapeutic strategy against cancer has increased considerably. Iron metabolism has been standing out as a target for the reprogramming of tumor-associated macrophages to M1-like phenotype with therapeutic purposes against cancer. Due to the importance of the iron levels in macrophage polarization states, iron oxide nanoparticles can be used to change the activation state of tumor-associated macrophages for a tumor suppressor phenotype and as an anti-tumor strategy.
    Keywords:  cancer; immunometabolism; immunotherapy; iron oxide nanoparticles; metabolic reprogramming of tumor-associated macrophages
    DOI:  https://doi.org/10.2217/nnm-2021-0255
  93. Front Endocrinol (Lausanne). 2021 ;12 765807
    Fluvastatin Reduces Glucose Tolerance in Healthy Young Individuals Independently of Cold Induced BAT Activity.
    Martina Felder, Claudia Irene Maushart, Gani Gashi, Jaël Rut Senn, Anton S Becker, Julian Müller, Miroslav Balaz, Christian Wolfrum, Irene A Burger, Matthias Johannes Betz.
      Background: Statins are commonly prescribed for primary and secondary prevention of atherosclerotic disease. They reduce cholesterol biosynthesis by inhibiting hydroxymethylglutaryl-coenzyme A-reductase (HMG-CoA-reductase) and therefore mevalonate synthesis. Several studies reported a small, but significant increase in the diagnosis of diabetes mellitus with statin treatment. The molecular mechanisms behind this adverse effect are not yet fully understood. Brown adipose tissue (BAT), which plays a role in thermogenesis, has been associated with a reduced risk of insulin resistance. Statins inhibit adipose tissue browning and have been negatively linked to the presence of BAT in humans. We therefore speculated that inhibition of BAT by statins contributes to increased insulin resistance in humans.Methods: A prospective study was conducted in 17 young, healthy men. After screening whether significant cold-induced thermogenesis (CIT) was present, participants underwent glucose tolerance testing (oGTT) and assessment of BAT activity by FDG-PET/MRI after cold-exposure and treatment with a β3-agonist. Fluvastatin 2x40mg per day was then administered for two weeks and oGTT and FDG-PET/MRI were repeated.
    Results: Two weeks of fluvastatin treatment led to a significant increase in glucose area under the curve (AUC) during oGTT (p=0.02), reduction in total cholesterol and LDL cholesterol (both p<0.0001). Insulin AUC (p=0.26), resting energy expenditure (REE) (p=0.44) and diet induced thermogenesis (DIT) (p=0.27) did not change significantly. The Matsuda index, as an indicator of insulin sensitivity, was lower after fluvastatin intake, but the difference was not statistically significant (p=0.09). As parameters of BAT activity, mean standard uptake value (SUVmean) (p=0.12), volume (p=0.49) and total glycolysis (p=0.74) did not change significantly during the intervention. Matsuda index, was inversely related to SUVmean and the respiratory exchange ratio (RER) (both R2 = 0.44, p=0.005) at baseline, but not after administration of fluvastatin (R2 = 0.08, p=0.29, and R2 = 0.14, p=0.16, respectively).
    Conclusions: Treatment with fluvastatin for two weeks reduced serum lipid levels but increased glucose AUC in young, healthy men, indicating reduced glucose tolerance. This was not associated with changes in cold-induced BAT activity.
    Keywords:  brown adipose tissue; cold-induced thermogenesis; diabetes; energy expenditure (EE); fluvastatin; glucose tolerance; statin
    DOI:  https://doi.org/10.3389/fendo.2021.765807
  94. Radiother Oncol. 2021 Nov 25. pii: S0167-8140(21)09004-6. [Epub ahead of print]
    Quality of life, HPV-status and phase angle predict survival in head and neck cancer patients under (chemo)radiotherapy undergoing nutritional intervention: Results from the prospective randomized HEADNUT-trial.
    Laura Magdalena Kutz, Jakob Abel, Diana Schweizer, Silke Tribius, Andreas Krüll, Cordula Petersen, Anastassia Löser.
      PURPOSE: To analyze the impact of quality of life (QoL), nutritional and clinical indicators on overall survival in patients with head and neck squamous cell cancer (HNSCC) undergoing (chemo)radiotherapy.MATERIALS AND METHODS: At the beginning, at the end of (chemo)radiotherapy and during follow-up, QoL was prospectively assessed using the EORTC-QLQ-C30 and -QLQ-H&N35 questionnaires. Data were analyzed in 58 out of 220 screened patients, who were randomized into a control and intervention group. All patients received a nutritional assessment including bioelectrical impedance analysis (BIA), laboratory testing, and a screening for malnutrition based on the questionnaires MUST, NRS-2002 and Nutriscore at baseline and at the end of therapy. The intervention consisted of an individualized nutritional counseling every 2 weeks.
    RESULTS: Except for emotional functioning, dyspnea, financial difficulties, dental problems and weight gain, all other scales from the EORTC-QLQ-C30 and -H&N35 deteriorated during (chemo)radiotherapy. At first follow-up, patients of the control group experienced more nausea and vomiting compared to those of the intervention group. (p=0.02). After performing a multivariable model, dental problems at the end of therapy (HR: 1.03; 95% CI: 1-1.06; p=0.03), HPV negativity (HR: 18.19, 95% CI: 1.61-204.17; p=0.02), and baseline phase angle (HR: 0.09; 95% CI: 0.01-0.82; p=0.03) were identified as predictors for overall survival.
    CONCLUSIONS: Factors influencing overall survival in patients with HNSCC undergoing (chemo)radiotherapy are complex and multifactorial. We were able to identify QoL-related (dental problems), clinical (HPV status) and nutritional (phase angle) factors as negative predictors for survival. This study was registered within the German Clinical Trials Register (DRKS00016862).
    Keywords:  HPV status; head and neck squamous cell cancer; malnutrition; phase angle; quality of life; survival
    DOI:  https://doi.org/10.1016/j.radonc.2021.11.011
  95. Curr Drug Metab. 2021 Dec 01.
    Identification and Quantification of MIDD0301 metabolites.
    M S Rashid Roni, Nicolas M Zahn, Brandon N Mikulsky, Daniel A Webb, Md Yeunus Mian, Daniel E Knutson, Margaret L Guthrie, James M Cook, Douglas C Stafford, Leggy A Arnold.
      BACKGROUND: MIDD0301 is an oral asthma drug candidate that binds GABAA receptors on airway smooth muscle and immune cells.OBJECTIVE: The objective of this study is to identify and quantify MIDD0301 metabolites in vitro and in vivo and determine the pharmacokinetics of oral, IP, and IV administrated MIDD0301.
    METHODS: In vitro conversion of MIDD0301 was performed using liver and kidney microsomes/S9 fractions followed by quantification using liquid chromatography-tandem mass spectrometry (LC-MS/MS). A LC-MS/MS method was developed using synthesized standards to quantify MIDD0301 and its metabolites in urine and feces. Blood, lung, and brain were harvested from animals that received MIDD0301 by oral, IP, and IV administration, followed by LCMS/MS quantification. Imaging mass spectrometry was used to demonstrate the presence of MIDD0301 in the lung after oral administration.
    RESULTS: MIDD0301 is stable in the presence of liver and kidney microsomes and S9 fractions for at least two hours. MIDD0301 undergoes conversion to the corresponding glucuronide and glucoside in the presence of conjugating cofactors. For IP and IV administration, unconjugated MIDD0301 together with significant amounts of MIDD0301 glucoside and MIDD0301 taurine were found in urine and feces. Less conjugation was observed following oral administration, with MIDD0301 glucuronide being the main metabolite. Pharmacokinetic quantification of MIDD0301 in blood, lung, and brain showed very low levels of MIDD0301 in the brain after oral, IV, or IP administration. The drug half-life in these tissues ranged between 4-6 hours for IP and oral and 1-2 hours for IV administration. Imaging mass spectrometry demonstrated that orally administered MIDD0301 distributes uniformly in the lung parenchyma.
    CONCLUSION: MIDD0301 undergoes no phase I and moderate phase II metabolism.
    Keywords:  GABAA receptor; MIDD0301; glucosidation; glucuronidation; imaging mass spectrometry; metabolism; taurine conjugate
    DOI:  https://doi.org/10.2174/1389200222666211202093841
  96. Thorax. 2021 Dec 01. pii: thoraxjnl-2021-217710. [Epub ahead of print]
    CT pectoralis muscle area is associated with DXA lean mass and correlates with emphysema progression in a tobacco-exposed cohort.
    Michael Emmet O'Brien, Richard H Zou, Nathan Hyre, Joseph K Leader, Carl R Fuhrman, Frank C Sciurba, Mehdi Nouraie, Jessica Bon.
      INTRODUCTION: Muscle loss is an important extrapulmonary manifestation of COPD. Dual energy X-ray absorptiometry (DXA) is the method of choice for body composition measurement but is not widely used for muscle mass evaluation. The pectoralis muscle area (PMA) is quantifiable by CT and predicts cross-sectional COPD-related morbidity. There are no studies that compare PMA with DXA measures or that evaluate longitudinal relationships between PMA and lung disease progression.METHODS: Participants from our longitudinal tobacco-exposed cohort had baseline and 6-year chest CT (n=259) and DXA (n=164) data. Emphysema was quantified by CT density histogram parenchymal scoring using the 15th percentile technique. Fat-free mass index (FFMI) and appendicular skeletal mass index (ASMI) were calculated from DXA measurements. Linear regression model relationships were reported using standardised coefficient (β) with 95% CI.
    RESULTS: PMA was more strongly associated with DXA measures than with body mass index (BMI) in both cross-sectional (FFMI: β=0.76 (95% CI 0.65 to 0.86), p<0.001; ASMI: β=0.76 (95% CI 0.66 to 0.86), p<0.001; BMI: β=0.36 (95% CI 0.25 to 0.47), p<0.001) and longitudinal (ΔFFMI: β=0.43 (95% CI 0.28 to 0.57), p<0.001; ΔASMI: β=0.42 (95% CI 0.27 to 0.57), p<0.001; ΔBMI: β=0.34 (95% CI 0.22 to 0.46), p<0.001) models. Six-year change in PMA was associated with 6-year change in emphysema (β=0.39 (95% CI 0.23 to 0.56), p<0.001) but not with 6-year change in airflow obstruction.
    CONCLUSIONS: PMA is an accessible measure of muscle mass and may serve as a useful clinical surrogate for assessing skeletal muscle loss in smokers. Decreased PMA correlated with emphysema progression but not lung function decline, suggesting a difference in the pathophysiology driving emphysema, airflow obstruction and comorbidity risk.
    Keywords:  emphysema; imaging/CT MRI etc; respiratory muscles; tobacco and the lung
    DOI:  https://doi.org/10.1136/thoraxjnl-2021-217710
  97. Oncogene. 2021 Dec 03.
    Oncogenic dependency on STAT3 serine phosphorylation in KRAS mutant lung cancer.
    Sultan Alhayyani, Louise McLeod, Alison C West, Jesse J Balic, Christopher Hodges, Liang Yu, Julian A Smith, Zdenka Prodanovic, Steven Bozinovski, Beena Kumar, Saleela M Ruwanpura, Mohamed I Saad, Brendan J Jenkins.
      The oncogenic potential of the latent transcription factor signal transducer and activator of transcription (STAT)3 in many human cancers, including lung cancer, has been largely attributed to its nuclear activity as a tyrosine-phosphorylated (pY705 site) transcription factor. By contrast, an alternate mitochondrial pool of serine phosphorylated (pS727 site) STAT3 has been shown to promote tumourigenesis by regulating metabolic processes, although this has been reported in only a restricted number of mutant RAS-addicted neoplasms. Therefore, the involvement of STAT3 serine phosphorylation in the pathogenesis of most cancer types, including mutant KRAS lung adenocarcinoma (LAC), is unknown. Here, we demonstrate that LAC is suppressed in oncogenic KrasG12D-driven mouse models engineered for pS727-STAT3 deficiency. The proliferative potential of the transformed KrasG12D lung epithelium, and mutant KRAS human LAC cells, was significantly reduced upon pS727-STAT3 deficiency. Notably, we uncover the multifaceted capacity of constitutive pS727-STAT3 to metabolically reprogramme LAC cells towards a hyper-proliferative state by regulating nuclear and mitochondrial (mt) gene transcription, the latter via the mtDNA transcription factor, TFAM. Collectively, our findings reveal an obligate requirement for the transcriptional activity of pS727-STAT3 in mutant KRAS-driven LAC with potential to guide future therapeutic targeting approaches.
    DOI:  https://doi.org/10.1038/s41388-021-02134-4
  98. J Proteome Res. 2021 Dec 01.
    Uncovering the Metabolic Mechanism of Salidroside Alleviating Microglial Hypoxia Inflammation Based on Microfluidic Chip-Mass Spectrometry.
    Fangfang Fan, Ning Xu, Yucheng Sun, Xuanhao Li, Xinchang Gao, Xizhen Yi, Yi Zhang, Xianli Meng, Jin-Ming Lin.
      Microglia are the main immune cells in the brain playing a critical role in neuroinflammation, and numerous pieces of evidence have proved that energy metabolism is closely associated with inflammation in activated microglia. Salidroside (Sal) isolated from Tibetan medicine Rhodiola crenulate can inhibit microglial hypoxia inflammation (HI). However, whether the inhibition is due to the intervening energy metabolic process in microglia is not clear. In this work, the hypoxic microenvironment of BV2 microglial cells was simulated using deferoxamine (DFO) in vitro and the change of cell metabolites (lactate, succinate, malate, and fumarate) was real-time online investigated based on a cell microfluidic chip-mass spectrometry (CM-MS) system. Meanwhile, for confirming the metabolic mechanism of BV2 cells under hypoxia, the level of HI-related factors (LDH, ROS, HIF-1α, NF-κB p65, TNF-α, IL-1β, and IL-6) was detected by molecular biotechnology. Integration of the detected results revealed that DFO-induced BV2 cell HI was associated with the process of energy metabolism, in which cell energy metabolism changed from oxidative phosphorylation to glycolysis. Furthermore, administration of Sal treatment could effectively invert this change, and two metabolites of Sal were identified: tyrosol and 4-hydroxyphenylacetic acid. In general, we illustrated a new mechanism of Sal for reducing BV2 cell HI injury and presented a novel analysis strategy that opened a way for real-time online monitoring of the energy metabolic mechanism of the effect of drugs on cells and further provided a superior strategy to screen natural drug candidates for HI-related brain disease treatment.
    Keywords:  hypoxia inflammation; metabolic mechanism; microfluidic chip-mass spectrometry; microglia; salidroside
    DOI:  https://doi.org/10.1021/acs.jproteome.1c00647
  99. Clin Nutr ESPEN. 2021 Dec;pii: S2405-4577(21)01075-5. [Epub ahead of print]46 305-313
    Metformin anticipates peak of lactate during high-intensity interval training but no changes performance or neuromuscular response in amateur swimmers.
    Matheus Silva Norberto, Tarine Botta De Arruda, Jonatas Augusto Cursiol, Vitor Luiz De Andrade, Gustavo Gomes De Araujo, Marcelo Papoti.
      BACKGROUND & AIMS: Metformin demonstrated potential to improve metabolic efficiency in short-intense and prolonged-continuous efforts. The present study investigates the acute effects of metformin intake on performance, rating of perceived exertion (RPE), blood lactate, blood glucose and neuromuscular parameters related to swimming high-intensity interval series.METHODS: A double-blind, crossover, randomized and placebo-controlled study was carried out. Seven healthy swimmers ingested metformin (500 mg) or placebo capsules on different days and performed a typical high-intensity training series (10 bouts of 50 m with a 3-min interval). Performance, RPE, neuromuscular parameters (lower and upper limbs), blood lactate and glucose were analyzed by the Wilcoxon Signed-Rank and Friedman's tests between supplementation situations and moments, respectively (p > 0.05), the moment where glucose and blood lactate peak were found were analyzed by a Student t-test (p > 0.05) and a Bayesian repeated-measures ANOVA for effects analysis (BFincl).
    RESULTS: The anticipation of blood glucose and peak lactate was signaling by the interaction effect (metformin increased and placebo decreased) between the eighth and the last bout (BFincl: 4.230 and 5.188 respectively). The second interaction effect of blood glucose and lactate (metformin maintained and placebo increased) during recoveries between 5 min and 7 min (BFincl: 3.825 and 3.806 respectively) also signaling the anticipatory behavior of both physiological parameters. The anticipation of blood lactate peak concentration after metformin intake confirms the anticipatory behavior of blood lactate (p: 0.015).
    CONCLUSIONS: The anticipatory behavior of glucose was not confirmed. Although the anticipatory peak of blood lactate, metformin does not affect neuromuscular responses, RPE and performance.
    REGISTRATION OF CLINICAL TRIAL: RBR-67wxdw8 Effects of metformin during swimmer training performance.
    Keywords:  Blood glucose; Blood lactate; Neuromuscular fatigue; Swimming; Typical series performance
    DOI:  https://doi.org/10.1016/j.clnesp.2021.09.739
  100. Oncol Lett. 2022 Jan;23(1): 19
    Use of oral glutamine in radiation-induced adverse effects in patients with thoracic and upper aerodigestive malignancies: Results of a prospective observational study.
    Amalia Papanikolopoulou, Nikolaos Syrigos, Louisa Vini, Maria Papasavva, Georgios Lazopoulos, Stelios Kteniadakis, Demetrios A Spandidos, Adrianni Charpidou, Nikolaos Drakoulis.
      Cancer growth in host tissues features glutamine (gln) depletion over time, decreasing epithelial cells' optimal functioning. In addition, radiotherapy (RT) and/or chemotherapy (CT) cause damage to normal tissues, probably enhanced by this depletion. The present study prospectively examined the effect of gln supplementation on 72 patients with thoracic and upper aerodigestive malignancies (T&UAM) treated with sequential or concurrent RT-CT or RT alone. All patients received prophylactic gln powder 15 g bid for the full duration of treatment. The severity of acute radiation toxicities was graded according to the RT Oncology Group/European Organization for Research and Treatment of Cancer criteria. Primary endpoints were the incidence of grade >2 toxicities, weight loss and requirement for analgesics, and the secondary endpoint was the association of the length of irradiated esophagus from treatment planning with the use of opioids. The incidence of adverse effects was as follows: Grade >2 stomatitis, 25.0%; esophagitis, 60.5%; dysphagia, 54.2%; pain, 25.4%; mycosis, 40.8%. Stomatitis grade >2 was more frequent in patients with head and neck tumors (P<0.001) and in those with prior surgery (P<0.001). Esophagitis (P=0.020) and dysphagia (P=0.008) grade >2 were more frequent in patients with concurrent RT-CT. Regarding analgesics, 9.9% of patients received no pain treatment, 56.3% received simple analgesic therapy and 33.8% opioids. Patients on opioid therapy had a greater mean length of irradiated esophagus (P=0.024) or length >12 cm (P=0.018). In 54.2% of patients, weight loss was observed, particularly with concurrent RT-CT (P=0.007). Thus, the use of oral gln may have an important role in reducing acute radiation toxicities and weight loss, and in lowering the requirement for analgesics in patients with T&UAM. Further randomized trials are required to identify the appropriate gln dose, duration of treatment and precise radiation dosimetric parameters in this group of patients. The present clinical trial was retrospectively registered in the ClinicalTrials.gov Protocol Registration and Results System (registration no. NCT05054517/22-09-2021).
    Keywords:  chest tumor; clinical study; glutamine; head and neck cancer; human subjects; lung cancer
    DOI:  https://doi.org/10.3892/ol.2021.13137
  101. Cancer Discov. 2021 Nov 30. pii: candisc.1077.2021. [Epub ahead of print]
    Mutant-IDH inhibits Interferon-TET2 signaling to promote immunoevasion and tumor maintenance in cholangiocarcinoma.
    Meng-Ju Wu, Lei Shi, Juan Dubrot, Joshua Merritt, Vindhya Vijay, Ting-Yu Wei, Emily Kessler, Kira E Olander, Ramzi Adil, Amaya Pankaj, Krishna Seshu Tummala, Vajira Weeresekara, Yuanli Zhen, Qibiao Wu, Meiqi Luo, William Shen, Maria Garcia-Beccaria, Mirian Fernandez-Vaquero, Christine Hudson, Sebastien Ronseaux, Yi Sun, Rodrigo Saad-Berreta, Russell W Jenkins, Tong Wang, Mathias Heikenwalder, Cristina R Ferrone, Lipika Goyal, Brandon Nicolay, Vikram Deshpande, Rahul M Kohli, Hongwu Zheng, Robert T Manguso, Nabeel Bardeesy.
      Isocitrate dehydrogenase 1 mutations (mIDH1) are common in cholangiocarcinoma. (R)-2-hydroxyglutarate generated by the mIDH1 enzyme inhibits multiple a-ketoglutarate-dependent enzymes, altering epigenetics and metabolism. Here, by developing mIDH1-driven genetically engineered mouse models, we show that mIDH1 supports cholangiocarcinoma tumor maintenance through an immunoevasion program centered on dual (R)-2-hydroxyglutarate-mediated mechanisms - suppression of CD8+ T cell activity and tumor cell-autonomous inactivation of TET2 DNA demethylase. Pharmacological mIDH1 inhibition stimulates CD8+ T cell recruitment and IFN-y expression and promotes TET2-dependent induction of IFN-y response genes in tumor cells. CD8+ T cell depletion or tumor cell-specific ablation of TET2 or Interferon-gamma receptor 1 causes treatment resistance. Whereas immune checkpoint activation limits mIDH1 inhibitor efficacy, CTLA4 blockade overcomes immunosuppression, providing therapeutic synergy. The findings in this mouse model of cholangiocarcinoma demonstrate that immune function and the IFN-y-TET2 axis are essential for response to mIDH1 inhibition and suggest a novel strategy for harnessing these inhibitors therapeutically.
    DOI:  https://doi.org/10.1158/2159-8290.CD-21-1077
  102. Clin Transl Med. 2021 11;11(11): e577
    Baicalein resensitizes tamoxifen-resistant breast cancer cells by reducing aerobic glycolysis and reversing mitochondrial dysfunction via inhibition of hypoxia-inducible factor-1α.
    Yan Chen, Jingyu Zhang, Minqin Zhang, Yuxuan Song, Yue Zhang, Shuangqin Fan, Shuang Ren, Lingyun Fu, Nenling Zhang, Hui Hui, Xiangchun Shen.
      Drug resistance is a major hurdle for the effectiveness of tamoxifen (TAM) to provide clinical benefit. Therefore, it is essential to identify a sensitizer that could be used to improve TAM efficacy in treating TAM-resistant breast cancer. Here, we investigated the ability of baicalein to reverse TAM resistance. We found that baicalein increased the efficacy of TAM in inhibiting proliferation and inducing apoptosis of TAM-resistant cells. It also enhanced the TAM-induced growth reduction of resistant cells from NOD/SCID mouse mammary fat pads, without causing obvious systemic toxicity. Analyses using the CellMiner tool and the Kaplan-Meier plotter database showed that HIF-1α expression was inversely correlated with TAM therapeutic response in NCI-60 cancer cells and breast cancer patients. HIF-1α expression was increased in TAM-resistant cells due to an increase in mRNA levels and reduced ubiquitin-mediated degradation. Baicalein reduced HIF-1α expression by promoting its interaction with PHD2 and pVHL, thus facilitating ubiquitin ligase-mediated proteasomal degradation and thereby suppressing the nuclear translocation, binding to the hypoxia-response element, and transcriptional activity of HIF-1α. As a result, baicalein downregulated aerobic glycolysis by restricting glucose uptake, lactate production, ATP generation, lactate/pyruvate ratio and expression of HIF-1α-targeted glycolytic genes, thereby enhancing the antiproliferative efficacy of TAM. Furthermore, baicalein interfered with HIF-1α inhibition of mitochondrial biosynthesis, which increased mitochondrial DNA content and mitochondrial numbers, restored the generation of reactive oxygen species in mitochondria, and thus enhanced the TAM-induced mitochondrial apoptotic pathway. The HIF-1α stabilizer dimethyloxallyl glycine prevented the baicalein-induced downregulation of glycolysis and mitochondrial biosynthesis and reduced the effects of baicalein on reversing TAM resistance. Our results indicate that baicalein is a promising candidate to help overcome TAM resistance by sensitizing resistant cells to TAM-induced growth inhibition and apoptosis. The mechanism underlying the effects of baicalein consists of inhibition of HIF-1α-mediated aerobic glycolysis and mitochondrial dysfunction.
    Keywords:  aerobic glycolysis; baicalein; hypoxia-inducible factor-1α; mitochondrial dysfunction; resistance; tamoxifen
    DOI:  https://doi.org/10.1002/ctm2.577
  103. J Cancer Res Ther. 2021 Nov;17(5): 1269-1274
    Computed tomography-guided cryoablation for adrenal metastases secondary to lung cancer.
    Wen-Long Zhang, Dong-Li Ruan, Li-Jun Sun, Tao Wang, Wei Zhang, Ya-Yong Huang.
      Objectives: The objective of the study was to assess the clinical efficacy of computed tomography (CT)-guided cryoablation as a means to treat adrenal metastasis (AM) secondary to lung cancer.Materials and Methods: This study was a single-center retrospective study that analyzed 39 consecutive patients with AM secondary to lung cancer who underwent CT-guided cryoablation in our center. The rates of complete ablation, local recurrence, local recurrence-free survival (RFS), and overall survival (OS) were analyzed.
    Results: The rates of primary and secondary complete ablation were 94.9% and 100%, respectively, and none of the patients suffered from a hypertensive crisis associated with the treatment. Over the follow-up period, 20.5% of the patients experienced local recurrence, and the median RFS duration was 26 months. The cumulative 1-, 3-, and 5-year local RFS rates in this study were 84.6%, 51.3%, and 5.9%, respectively. Extra-adrenal gland metastases were detected in five patients. Over the course of follow-up, 26 patients died. The mean OS duration was 34 months with cumulative 1-, 3-, and 5-year OS rates of 89.7%, 53.4%, and 8.3%, respectively. Advanced age (P = 0.001), primary adenocarcinoma (P = 0.006), other primary lung cancers (P = 0.038), and primary Stage III lung cancers (P = 0.007) were all found to be independent predictive factors of poor OS in these patients.
    Conclusion: CT-guided cryoablation can be safely and effectively used to control AM secondary to lung cancer, and patients with AM secondary to lung squamous cell carcinoma may be best suited for this form of treatment.
    Keywords:  Adrenal; cryoablation; lung cancer; metastasis; survival
    DOI:  https://doi.org/10.4103/jcrt.JCRT_1520_20
  104. BMC Pulm Med. 2021 Dec 03. 21(1): 395
    FeV1 and BMI influence King's Sarcoidosis Questionnaire score in sarcoidosis patients.
    Björn Christian Frye, Laura Potasso, Erik Farin-Glattacker, Surrinder Birring, Joachim Müller-Quernheim, Jonas Christian Schupp.
      BACKGROUND: Sarcoidosis is granulomatous disease of unknown origin affecting organ function and quality of life. The King's Sarcoidosis Questionnaire (KSQ) serves as a tool to assess quality of life in sarcoidosis patients with general health and organ specific domains. A German translation has been validated in a German cohort. In this study we assessed, whether clinical parameters influence KSQ scores.METHODS: Clinical data (e.g. lung function, organ impairment, serological parameters) for the German validation cohort were extracted from clinical charts and investigated by correlation and linear regression analyses.
    RESULTS: KSQ subdomain scores were generally lower in patients with respective organ manifestation or on current therapy. LUNG subdomain was significantly predicted by lung functional parameters, however for general health status, only FeV1 exerted significant influence. GHS was not influenced by serological parameters, but was significantly negatively correlated with body mass index (BMI). KSQ provides additional information beyond lung function, clinical or serological parameters in sarcoidosis patients. Notably, high BMI is significantly negatively associated with patients' well-being as measured by KSQ-GHS.
    CONCLUSION: This observation may direct further studies investigating the effect of obesity on sarcoidosis-related quality of life and strategies to intervene with steroid-sparing therapies and measures of life style modifications. Trial registration This study was registered in the German Clinical Trials Register (reference number DRKS00010072). Registered January 2016.
    Keywords:  King’s Sarcoidosis Questionnaire; Quality of life; Sarcoidosis
    DOI:  https://doi.org/10.1186/s12890-021-01761-7
  105. Cancer Cell Int. 2021 Nov 27. 21(1): 631
    NEDD4L-induced ubiquitination mediating UBE2T degradation inhibits progression of lung adenocarcinoma via PI3K-AKT signaling.
    Yongbing Chen, Haihua Hong, Qingqing Wang, Junqiang Li, Wenfeng Zhang, Tingting Chen, Pu Li.
      BACKGROUND: A number of studies have indicated that Ubiquitin-conjugating enzyme E2T (UBE2T), as an oncogene, promotes progression and metastasis of lung cancer, including lung adenocarcinoma (LUAD), but it is completely unknown whether and how UBE2T is ubiquitylated and degraded, and by which E3 ligase. NEDD4L plays a critical role in the regulation of cellular processes of various cancers, most of which is attributed to its E3 ubiquitin ligase function. However, the relationship between NEDD4L and UBE2T in LUAD has not been elucidated.METHODS: The relationship between NEDD4L and UBE2T in LUAD tissues and cells was found by bioinformatic analyses and immunoblotting. Cell counting kit-8, colony formation assay, half-life analysis and the in vivo ubiquitylation assay, generation of xenograft model were performed to determine how NEDD4L regulates UBE2T and its downstream signaling pathway in vitro and in vivo.
    RESULTS: Bioinformatic analyses found that NEDD4L, as a potential correlation E3 ligase of UBE2T, was negatively correlated with UBE2T in LUAD. Consistently, UBE2T protein half-life was shortened or extended by NEDD4L overexpression or depletion, respectively. NEDD4L inhibited LUAD cell progression in vitro and in vivo via inducing the ubiquitination-mediated UBE2T degradation, which repressed PI3K-AKT signaling. Similarly, NEDD4L predicted a better patient survival, whereas UBE2T predicted a worse survival.
    CONCLUSIONS: Collectively, our results reveal that NEDD4L is a novel E3 ligase of UBE2T, which can inhibit PI3K-AKT signaling by targeting for UBE2T ubiquitination and degradation, resulting in repression of LUAD cell progression.
    Keywords:  Lung adenocarcinoma; NEDD4L; PI3K-AKT signaling; UBE2T; Ubiquitylation
    DOI:  https://doi.org/10.1186/s12935-021-02341-9
  106. Cell Death Dis. 2021 Nov 27. 12(12): 1108
    SLC25A1 promotes tumor growth and survival by reprogramming energy metabolism in colorectal cancer.
    Ying Yang, Jiaxing He, Bo Zhang, Zhansheng Zhang, Guozhan Jia, Shiqi Liu, Tao Wu, Xianli He, Nan Wang.
      Abnormal lipid metabolism has been commonly observed in various human cancers, including colorectal cancer (CRC). The mitochondrial citrate carrier SLC25A1 (also known as mitochondrial citrate/isocitrate carrier, CIC), has been shown to play an important role in lipid metabolism regulation. Our bioinformatics analysis indicated that SLC25A1 was markedly upregulated in CRC. However, the role of SLC25A1 in the pathogenesis and aberrant lipid metabolism in CRC remain unexplored. Here, we found that SLC25A1 expression was significantly increased in tumor samples of CRC as compared with paired normal samples, which is associated with poor survival in patients with CRC. Knockdown of SLC25A1 significantly inhibited the growth of CRC cells by suppressing the progression of the G1/S cell cycle and inducing cell apoptosis both in vitro and in vivo, whereas SLC25A1 overexpression suppressed the malignant phenotype. Additionally, we demonstrated that SLC25A1 reprogrammed energy metabolism to promote CRC progression through two mechanisms. Under normal conditions, SLC25A1 increased de novo lipid synthesis to promote CRC growth. During metabolic stress, SLC25A1 increased oxidative phosphorylation (OXPHOS) to protect protects CRC cells from energy stress-induced cell apoptosis. Collectively, SLC25A1 plays a pivotal role in the promotion of CRC growth and survival by reprogramming energy metabolism. It could be exploited as a novel diagnostic marker and therapeutic target in CRC.
    DOI:  https://doi.org/10.1038/s41419-021-04411-2
  107. Clin Nutr ESPEN. 2021 Dec;pii: S2405-4577(21)00327-2. [Epub ahead of print]46 33-39
    Probiotic supplementation improves clinical outcomes and quality of life indicators in patients with plaque psoriasis: A randomized double-blind clinical trial.
    Jalal Moludi, Hamed Khedmatgozar, Somayeh Saiedi, Hamidreza Razmi, Mohammad Alizadeh, Behzad Ebrahimi.
      BACKGROUND & AIMS: The impact of probiotics on psoriasis, a systemic inflammatory disease, remains obscure, thus we decided to evaluate quality of life (QOL), oxidative stress, inflammatory markers and clinical outcome in psoriasis patients.METHODS: Fifty patients with plaque psoriasis randomized into two groups, group 1 received probiotic drink with Lactobacillus strains for 8 weeks while group 2 haven't received any probiotic supplements at this period. The Dermatology Life Quality Index (DLQI) and Beck's questionnaire (BDI) were used to investigate the quality of life and depression, respectively. The effects of supplementation on malondialdehyde (MDA), hs-CRP, IL-6, total antioxidant capacity (1) and psoriasis area and severity index (PASI), psoriasis symptom scale (PSS) were measured at the beginning of the study and after week 8th.
    RESULTS: Total BDI scores significantly improved in the probiotic group in comparison with the placebo group (-6.15 ± 2.10 vs. 1.39 ± 1.80, P = 0.017) and DLQI (-9.50 ± 4.1 vs. 0.12 ± 0.6, P = 0.045) as well. Group 1 had a considerable reduction in PASI and PSS scores compared to the placebo group (-5.26 ± 3.75 vs. 0.48 ± 1.37, P = 0.049) and (-4.85 ± 3.10 vs. 0.43 ± 0.80, P = 0.047), respectively. In addition, the intervention group have shown increase in TAC levels (45.99 ± 23.33 vs -13.54 ± 30.7 mmol/L, P = 0.030), and decrease in hs-CRP levels (-1.55 ± 0.85 vs. -0.49 + 0.27 mg/L, P = 0.015), IL-6 levels (-4.04 ± 1.30 vs. -1.50 + 0.38 mg/L, P = 0.050) and MDA levels (-71.08 ± 35.73 vs. -9.8 + 15.6 nmol/mL, P = 0.013) compared to the placebo group.
    CONCLUSIONS: Probiotics improve patients' quality of life and inflammatory biomarkers in psoriatic patients. Further studies are mandatory to propose probiotics as routinely prescribed therapy in inflammatory dermatoses.
    TRIAL REGISTRATION: This trial was registered at www.irct.ir as IRCT20180712040438N2.
    Keywords:  Clinical outcome; Oxidative stress; Probiotic; Psoriasis; Quality of life
    DOI:  https://doi.org/10.1016/j.clnesp.2021.09.004
  108. Exp Cell Res. 2021 Nov 27. pii: S0014-4827(21)00508-5. [Epub ahead of print]410(1): 112952
    Fermentation supernatant of Staphylococcus aureus drives catabolism in chondrocytes via NF-κB signaling mediated increase of cholesterol metabolism.
    Qian Wang, Jiefeng Huang, Shuaijun Li, Yun Zhang, Ruizhu Sun, Jiaxin Ren, Si Shi, Lei Cui.
      Septic arthritis induced by Staphylococcus aureus (S. aureus) causes irreversible cartilage degradation and subsequent permanent joint dysfunction. Recently, cartilage degradation in osteoarthritis is recognized to be associated with metabolic disorders. However, whether cholesterol metabolism is linked to septic arthritis pathology remains largely unknown. Here, we found that exposure to fermentation supernatant (FS) of S. aureus in chondrocytes resulted in a significant increase in expression of key modulators involved in cholesterol metabolism, including lectin-type oxidized low density lipoprotein receptor 1 (LOX1), cholesterol 25-hydroxylase (CH25H), 25- hydroxycholesterol 7α-hydroxylase (CYP7B1) as well as retinoic acid-related orphan receptor alpha (RORα), a binding receptor for cholesterol metabolites. We further demonstrated that enhancement of CH25H/CYP7B1/RORα axis resulted from FS exposure was mediated by activation of NF-κB signaling, along with upregulation in catabolic factors including matrix metallopeptidases (MMP3 and MMP13), aggrecanase-2 (ADAMTS5), and nitric oxide synthase-2 (NOS2) in chondrocytes. Exogenous cholesterol acts synergistically with FS in activating NF-κB pathway and increases cholesterol metabolism. While, the addition of tauroursodeoxycholic acid (TUDCA) which promotes cholesterol efflux, resulted in remarkable reduction of intracellular cholesterol level and restoration of balance between anabolism and catabolism in FS treated chondrocytes. Collectively, our data indicated that, in response to FS of S. aureus, NF-κB signaling activation coupled with increased cholesterol metabolism to stimulate catabolic factors in chondrocytes, highlighting cholesterol metabolism as a potential therapeutic target for treating septic arthritis.
    Keywords:  Cholesterol metabolism; Chondrocytes; Nuclear factor-kappa B; Septic arthritis; Staphylococcus aureus; TUDCA
    DOI:  https://doi.org/10.1016/j.yexcr.2021.112952
  109. Clin Nutr ESPEN. 2021 Dec;pii: S2405-4577(21)01088-3. [Epub ahead of print]46 216-222
    Nutritional and prognostic significance of abdominal wall thickness measured during percutaneous endoscopic gastrostomy in older individuals with dysphagia.
    Shigenori Masaki, Takashi Kawamoto.
      BACKGROUND & AIMS: The significance of abdominal wall thickness (AWT) for nutritional assessment remains unclear. This study aimed to evaluate the nutritional and prognostic significance of AWT measured during percutaneous endoscopic gastrostomy (PEG) in older patients with dysphagia.METHODS: This single-center retrospective cohort study enrolled older adults with dysphagia who underwent PEG for enteral nutrition using the introducer technique between February 2010 and January 2019. We examined the association between AWT measured during PEG and nutritional status items, including body mass index (BMI), serum albumin (Alb), total lymphocyte count (TLC), total cholesterol (TC), hemoglobin (Hb), and C-reactive protein (CRP). The shaft length of the PEG tube inserted, which is an approximation value of the AWT, was used for statistical analysis. Patients were divided into three groups: low-AWT group (shaft length ≤2.5 cm), medium-AWT group (shaft length 3.0-3.5 cm), and high-AWT group (shaft length ≥4.0 cm). We performed the Kruskal-Wallis test and multiple linear regression analysis with multiple imputation. We performed survival analysis using the Kaplan-Meier method, log-rank test, and Cox regression analysis.
    RESULTS: A total of 520 patients were identified: 115, 258, and 147 patients in the low-AWT, medium-AWT, and high-AWT groups, respectively. Higher AWT was significantly associated with higher BMI, Alb, TLC, TC, Hb, and lower CRP levels. Multiple linear regression analysis revealed that BMI and TLC were significant predictors of AWT (BMI: coefficient 1.16E-01, 95% confidence interval [CI], 9.77E-02-1.33E-01, P < 0.001; TLC: coefficient 1.18E-04, 95% CI, 2.72E-05-2.09E-04, P = 0.011). The median survival time was the longest in the high-AWT group (low-AWT, 359 days; medium-AWT, 851 days; high-AWT, 1662 days; P < 0.001). The hazard ratio for the high-AWT group relative to the medium-AWT group was 0.59 (95% CI, 0.41-0.85, P = 0.004), and that relative to the low-AWT group was 0.34 (95% CI, 0.24-0.51, P < 0.001).
    CONCLUSIONS: Higher AWT was associated with better nutritional status and survival. AWT may help assess nutritional status and predict survival in older dysphagic patients with PEG.
    Keywords:  Abdominal wall thickness; Nutrition; Nutritional assessment; Older people; Percutaneous endoscopic gastrostomy; Survival
    DOI:  https://doi.org/10.1016/j.clnesp.2021.10.005
  110. Epidemiol Prev. 2021 Sep-Oct;45(5):45(5): 353-367
    L'impatto dell'esposizione occupazionale ad amianto sul tumore del polmone in Italia.
    Manuela De Sario, Lisa Bauleo, Corrado Magnani, Daniela Ferrante, Alessandro Marinaccio, Paola Michelozzi, Elisa Romeo.
      OBJECTIVES: to perform a meta-analysis of cohort studies on lung cancer mortality in occupational sectors exposed to asbestos, particularly in the construction sector, and to use data from Italian cohorts exposed to asbestos to estimate the number of lung cancer cases attributable to asbestos in Italy.METHODS: systematic literature review and estimation of lung cancer deaths and cases attributable to asbestos in Italian cohorts and from the Italian National Register of Malignant Mesothelioma (ReNaM).
    SETTING AND PARTICIPANTS: the literature search was conducted in Medline and Embase (Ovid), including papers published from 1999 to May 2019. The following sectors were considered most exposed to asbestos: asbestos-cement, rolling-stock, shipyards, dockyards, glass workers, insulators, asphalt roll production workers, industrial ovens, miners. Moreover, the construction sector was included.
    MAIN OUTCOME MEASURES: the standardized mortality ratio (SMR) was estimated from the meta-analysis of the literature review. The ratio lung cancer to mesothelioma attributable cases was estimated by occupational sector from the Italian cohorts. For the construction sector, the ratio lung cancer to mesothelioma cases was estimated within the exposed workers estimated by CAREX (1990-1993). The ratios were applied to the mesothelioma cases registered at the ReNaM for the 2010-2015 period, to obtain a national estimate of lung cancer cases attributable to occupational exposure to asbestos.
    RESULTS: the meta-analytical SMR for lung cancer in men varied between 1.05 (asphalt roll) and 2.36 (insulation). The mean risk for all sectors was 1.37 in men and 1.60 in women. It increased in cohorts with latency higher than 20 years. Significant risks were observed in asbestos-cement (both genders), construction, and mining sectors. There was a mean of 1.1, 2.7, and 2.8 lung cancer deaths per mesothelioma death in the cement-asbestos, harbour, and construction sectors, respectively. The impact in terms of lung cancer cases estimated at the national level was equal to 3,814 cases between 2010 and 2015.
    CONCLUSIONS: to provide an overall assessment of the impact of the occupational asbestos exposure, it is important to consider lung cancer cases, in addition to malignant mesotheliomas. This study was able to estimate the impact of asbestos on lung cancer in Italy 25 years after the ban of this occupational carcinogen, with the largest contribution in terms of attributable cases coming from the construction sector. It is urgent to implement adequate information and prevention strategies, health surveillance of workers, and the appropriate legal framework for insurance purposes.
    Keywords:  Asbestos; Attributable proportion; Cohorts; Meta-analysis; Occupation
    DOI:  https://doi.org/10.19191/EP21.5.P353.102
  111. J Cancer Res Ther. 2021 Nov;17(5): 1281-1285
    Roles of hydroxyacyl-CoA dehydrogenase trifunctional multienzyme complex subunit alpha, a lipid metabolism enzyme, in Wilms tumor patients.
    Xiangyu Wu, Run Feng, Xiaoqing Wang, Feng Guo, Wei Liu.
      Objectives: Wilms tumor is a common pediatric malignant tumor that accounts for approximately 95% of kidney tumors in children. The role of lipid metabolism in tumors has attracted increased attention in recent years. We examined the role of hydroxyacyl-CoA dehydrogenase trifunctional multienzyme complex subunit alpha (HADHA), a lipid metabolism enzyme, in the pathogenesis of Wilms tumor.Materials and Methods: In a previous study, we screened Wilms tumors and adjacent normal tissues for differentially expressed proteins by mass spectrometry and verified the results by western blot analysis. The Oncomine database and quantitative reverse transcription-polymerase chain reaction were used to verify the expression of HADHA at the genetic level. Immunohistochemistry and immunofluorescence were also used to validate the differential expression of the HADHA protein. The relationship between histopathological typing, clinical pathology, and HADHA expression was analyzed in 65 paraffin-embedded specimens from pediatric Wilms tumor patients. Kaplan-Meier survival curves were used to analyze the relationship between the expression of HADHA and patient prognosis.
    Results: HADHA was expressed at low levels in Wilms tumor tissue compared with the corresponding normal tissue. The expression of HADHA was closely associated with histopathological typing (P = 0.030). The prognostic analysis of 65 children with Wilms tumor showed that high expression of HADHA was closely associated with poor prognosis (P = 0.046).
    Conclusions: HADHA expression is downregulated in Wilms tumor tissues, but high expression in tumor tissues is associated with clinical stage and the prognosis of children with this tumor.
    Keywords:  Clinicopathological parameters; Wilms tumor; hydroxyacyl-CoA dehydrogenase trifunctional multienzyme complex subunit alpha; lipid metabolism; prognosis
    DOI:  https://doi.org/10.4103/jcrt.jcrt_1388_21
  112. Am J Respir Cell Mol Biol. 2021 Dec 01.
    Human Rhinovirus Infection of the Respiratory Tract Affects Sphingolipid Synthesis.
    Emily Wasserman, Rika Gomi, Anurag Sharma, Seunghee Hong, Rohan Bareja, Jinghua Gu, Uthra Balaji, Arul Veerappan, Benjamin I Kim, Wenzhu Wu, Andrea Heras, Jose Perez-Zoghbi, Biin Sung, Seyni Gueye-Ndiaye, Tilla S Worgall, Stefan Worgall.
      The 17q21 asthma susceptibility locus includes asthma risk alleles associated with decreased sphingolipid synthesis, likely resulting from increased expression of ORMDL3. ORMDL3 inhibits serine-palmitoyl transferase (SPT), the rate limiting enzyme of de novo sphingolipid synthesis. There is evidence that decreased sphingolipid synthesis is critical to asthma pathogenesis. Children with asthma and 17q21 asthma risk alleles display decreased sphingolipid synthesis in blood cells. Reduced SPT activity results in airway hyperreactivity, a hallmark feature of asthma. 17q21 asthma risk alleles are also linked to childhood infections with human rhinovirus (RV). This study evaluates the interaction of RV with the de novo sphingolipid synthesis pathway, and the alterative effects of concurrent SPT inhibition in SPT-deficient mice and human airway epithelial cells. In mice, RV infection shifted lung sphingolipid synthesis gene expression to a pattern that resembles genetic SPT deficiency, including decreased expression of Sptssa, a small SPT subunit. This pattern was pronounced in lung EpCAM+ epithelial cells and reproduced in human bronchial epithelial cells. RV did not affect Sptssa expression in lung CD45+ immune cells. RV increased sphingolipids unique to the de novo synthesis pathway in mouse lung and human airway epithelial cells. Interestingly, these de novo sphingolipid species were reduced in the blood of RV infected, wild-type mice. RV exacerbated SPT-deficiency-associated airway hyperreactivity. Airway inflammation was similar in RV-infected wild-type and SPT deficient mice. This study reveals the effects of RV infection on the de novo sphingolipid synthesis pathway, elucidating a potential mechanistic link between 17q21 asthma risk alleles and rhinoviral infection.
    Keywords:  17q21; asthma; early childhood wheeze; rhinovirus; sphingolipids
    DOI:  https://doi.org/10.1165/rcmb.2021-0443OC
  113. Int J Chron Obstruct Pulmon Dis. 2021 ;16 3189-3199
    Lower Prevalence of Osteoporosis in Patients with COPD Taking Anti-Inflammatory Compounds for the Treatment of Diabetes: Results from COSYCONET.
    COSYCONET Study Group
      Background: Patients with chronic obstructive pulmonary disease (COPD) often have osteoporosis and diabetes as comorbid conditions. Anti-diabetic medication, including metformin, has protective effects on osteoporosis in experimental studies. We therefore studied whether patients with COPD receiving anti-diabetic medication had a lower osteoporosis prevalence in a large COPD cohort, COSYCONET.Methods: Assessment of osteoporosis was based on patients' reports of physician-based diagnoses and the presence of disease-specific medication. The predictive value of physical characteristics, lung function, comorbidities, cardiovascular medication, and the use of anti-inflammatory diabetes medication, including metformin, sulfonylureas, glinides or DPP4I, was evaluated using logistic regression analysis. ClinicalTrials.gov: NCT01245933.
    Results: In total, 2222 patients were eligible for analysis (863 [39%] female, mean age 65 y), 515 of whom had higher symptoms and exacerbations (Global Initiative for Chronic Obstructive Lung Disease group D). Osteoporosis was present in 15.8% of the overall cohort, and in 24.1% of GOLD D patients. Regression analyses identified the following as associated with osteoporosis (p < 0.05): female sex, higher age, lower body-mass index, asthma, higher air trapping, oral steroids, and cardiovascular medication. Although oral anti-diabetic medication was overall not associated with a lower prevalence of osteoporosis (p = 0.131), anti-inflammatory anti-diabetic medication (p = 0.009) and metformin-containing therapy (p = 0.039) were. This was driven by GOLD D patients.
    Conclusion: In a large COPD cohort, anti-inflammatory diabetes therapy, including metformin, was associated with a lower prevalence of osteoporosis, especially in patients with higher symptoms and exacerbations. These findings suggest a protective effect of common anti-diabetic medication on osteoporosis, possibly as a result of attenuated systemic inflammation.
    Keywords:  anti-inflammatory; chronic obstructive pulmonary disease; diabetes; inhaled corticosteroids; metformin; oral corticosteroids
    DOI:  https://doi.org/10.2147/COPD.S335029
  114. Pol Arch Intern Med. 2021 Dec 01.
    Profound neutropenia related to consolidation durvalumab following concurrent radiochemotherapy in III stage non-small-cell lung cancer patient.
    Artur Bandura, Michał Kurlapski, Małgorzata Kamińska, Rafał Dziadziuszko, Jacek Jassem.
      
    DOI:  https://doi.org/10.20452/pamw.16161
  115. Zhonghua Wei Zhong Bing Ji Jiu Yi Xue. 2021 Sep;33(9): 1145-1148
    [Advances in research on the relationship between changes in cellular metabolism and lung diseases].
    Menglu Xu, Yiding Zuo, Li Zhou, Chunling Jiang.
      As the place for gas exchange, the lungs are metabolically active, and their energy consumption are essential for regulating common cell functions and maintaining the unique function of lung tissues to synthesize pulmonary surfactants. The metabolic pathways of pulmonary cells mainly include glycolysis, pentose phosphate pathway, and tricarboxylic acid cycle. Recent studies have found that changes in pulmonary cells metabolism are closely related to a variety of lung diseases. Herein, we review the main pathways of pulmonary cells metabolism and the relationship between changes in cell metabolism and the four lung diseases of chronic obstructive pulmonary disease (COPD), asthma, idiopathic pulmonary fibrosis (IPF), and pulmonary hypertension (PH), to find new ways to treat lung diseases.
    DOI:  https://doi.org/10.3760/cma.j.cn121430-20210119-00081
  116. Clin Med (Lond). 2021 Nov;21(6): e578-e583
    Lung cancer for the non-respiratory physician.
    Emma O'Dowd, Joe Mackenzie, Haval Balata.
      Lung cancer continues to be the leading cause of cancer death globally. Delayed diagnosis is a major contributing factor to poor outcomes and remains a key challenge to overcome. While debate around the implementation of lung cancer screening for asymptomatic high-risk individuals continues, rapid access to relevant diagnostic tests is essential. The new National Optimal Lung Cancer Pathway describes 'diagnostic standards of care' in an effort to implement best practice, reduce variation and improve delays in diagnosis, staging and treatment of lung cancer. Lung cancer treatment continues to develop with new surgical techniques, radiotherapy options and more drugs being licensed as part of standard treatment. We provide an overview of the core lung cancer diagnostic steps, recognition and management of acute presentations as well as the latest treatment options.
    Keywords:  DSOCs; diagnostic standards of care; diagnostics; lung cancer
    DOI:  https://doi.org/10.7861/clinmed.2021-0657
  117. Comput Struct Biotechnol J. 2021 ;19 5946-5959
    Synergistic alterations in the multilevel chromatin structure anchor dysregulated genes in small cell lung cancer.
    Dan Guo, Qiu Xie, Shuai Jiang, Ting Xie, Yaru Li, Xin Huang, Fangyuan Li, Tingting Wang, Jian Sun, Anqi Wang, Zixin Zhang, Hao Li, Xiaochen Bo, Hebing Chen, Zhiyong Liang.
      Small cell lung cancer (SCLC) is an aggressive form of lung cancer that uniquely changes the chromosomal structure, although the basis of aberrant gene expression in SCLC remains largely unclear. Topologically associated domains (TADs) are structural and functional units of the human genome. Genetic and epigenetic alterations in the cancer genome can lead to the disruption of TAD boundaries and may cause gene dysregulation. To understand the potential regulatory role of this process in SCLC, we developed the TAD boundary alteration-related gene identification in tumors (TARGET) computational framework, which enables the systematic identification of candidate dysregulated genes associated with altered TAD boundaries. Using TARGET to compare gene expression profiles between SCLC and normal human lung fibroblast cell lines, we identified >100 genes in this category, of which 24 were further verified in samples from patients with SCLC using NanoString. The analysis revealed synergistic chromatin structure alteration at the A/B compartment and TAD boundary levels that underlies aberrant gene expression in SCLC. TARGET is a novel and powerful tool that can be used to explore the relationship of chromatin structure alteration to gene dysregulation related to SCLC tumorigenesis, progression, and prognosis.
    Keywords:  Hi-C; NanoString; SCLC; TAD; TARGET; Tumorigenesis
    DOI:  https://doi.org/10.1016/j.csbj.2021.11.003
  118. Cell Death Discov. 2021 Nov 29. 7(1): 369
    LncRNA SFTA1P mediates positive feedback regulation of the Hippo-YAP/TAZ signaling pathway in non-small cell lung cancer.
    Bowen Zhu, Megan Finch-Edmondson, Kim Whye Leong, Xiaoqian Zhang, Mitheera V, Quy Xiao Xuan Lin, Yaelim Lee, Wei Ting Ng, Huili Guo, Yue Wan, Marius Sudol, Ramanuj DasGupta.
      Long non-coding RNAs (lncRNAs) regulate numerous biological processes involved in both development and carcinogenesis. Hippo-YAP/TAZ signaling, a critical pathway responsible for organ size control, is often dysregulated in a variety of cancers. However, the nature and function of YAP/TAZ-regulated lncRNAs during tumorigenesis remain largely unexplored. By profiling YAP/TAZ-regulated lncRNAs, we identified SFTA1P as a novel transcriptional target and a positive feedback regulator of YAP/TAZ signaling. Using non-small cell lung cancer (NSCLC) cell lines, we show that SFTA1P is transcriptionally activated by YAP/TAZ in a TEAD-dependent manner. Functionally, knockdown of SFTA1P in NSCLC cell lines inhibited proliferation, induced programmed cell death, and compromised their tumorigenic potential. Mechanistically, SFTA1P knockdown decreased TAZ protein abundance and consequently, the expression of YAP/TAZ transcriptional targets. We provide evidence that this phenomenon could potentially be mediated via its interaction with TAZ mRNA to regulate TAZ translation. Our results reveal SFTA1P as a positive feedback regulator of Hippo-YAP/TAZ signaling, which may serve as the molecular basis for lncRNA-based therapies against YAP/TAZ-driven cancers.
    DOI:  https://doi.org/10.1038/s41420-021-00761-0
  119. Sichuan Da Xue Xue Bao Yi Xue Ban. 2021 Nov;52(6): 923-928
    [Research Progress in Glucose Metabolism of Chondrocytes].
    Jie-Ya Wei, De-Mao Zhang, Jing Xie, Xue-Dong Zhou.
      Chondrocytes have a limited supply of glucose and oxygen for metabolism since articular cartilages are relatively avascular. We herein reviewed the characteristics of chondrocyte glucose metabolism and the new research progress in chondrocyte glucose metabolism in the osteoarthritis process. Current research has shown that chondrocytes obtain glucose from synovial fluids and subchondral bones, take in glucose via specific glucose transporters, and metabolize glucose mainly through glycolysis and mitochondrial respiration to produce adenosine triphosphate (ATP). Glucose metabolism in chondrocytes is distinctive because it relies much more on glycolysis rather than mitochondrial respiration for ATP production, and shows Warburg effect and Crabtree effect. In osteoarthritic chondrocytes, the glucose metabolism disorder is presented as further suppression of mitochondrial respiration, over-active or impaired glycolysis, and decreased total production of ATP. However, the significance of the glucose supply for chondrocytes from synovial fluids and subchondral bones remains undefined. There are still disputes in the understanding of the changes in glycolytic pathways in osteoarthritic chondrocytes. Therefore, future research is needed to explore the characteristics of glucose metabolism in normal and osteoarthritic chondrocytes in order to develop new diagnostic and therapeutic strategies for osteoarthritis.
    Keywords:  Chondrocytes; Glucose metabolism; Glycolysis; Osteoarthritis
    DOI:  https://doi.org/10.12182/20211160206
  120. Eur J Cancer. 2021 Nov 30. pii: S0959-8049(21)01182-5. [Epub ahead of print]
    Pharmacokinetic and pharmacogenomic analysis of low-dose afatinib treatment in elderly patients with EGFR mutation-positive non-small cell lung cancer.
    Hidenori Mizugaki, Satoshi Oizumi, Yuka Fujita, Toshiyuki Harada, Yoshiro Nakahara, Taichi Takashina, Ryo Ko, Kageaki Watanabe, Takamasa Hotta, Hiroyuki Minemura, Sho Saeki, Hajime Asahina, Keiichi Nakamura, Hiromi Nakamura, Fumie Hosoda, Shigehiro Yagishita, Akinobu Hamada.
      PURPOSE: An increasing number of advanced non-small cell lung cancer (NSCLC) cases are being reported in the ageing population. However, studies on the use of afatinib in elderly patients are scarce. We conducted a prospective multicentre, single-arm, and open-label phase II trial for low-dose afatinib (30 mg/day) use in elderly patients with NSCLC with EGFR mutation to assess quality-of-life (QOL) and pharmacokinetic (PK)/pharmacogenomic (PGx) parameters.PATIENTS AND METHODS: The primary end-point was the objective response rate (ORR), and the planned number of registered cases was 35, with a threshold ORR of 50%, an expected ORR of 75%, α of 0.05, and β of 0.1. Secondary end-points were progression-free survival (PFS), overall survival (OS), the incidence rate of adverse events (AEs), QOL survey (FACT-L), and trough plasma concentration of afatinib at steady state (Css) and at the occurrence of clinically significant AEs.
    RESULTS: The median age of the patients was 79 years. The ORR was 80.0% and the disease control rate was 91.4%. The median PFS and OS were 15.6 and 29.5 months, respectively. Four patients discontinued because of AEs. Treatment-related death was not observed. No significant change in QOL was observed at baseline and after 4, 8, and 12 weeks. Css was comparable with those in previous reports and was significantly higher in patients with grade 3 AEs. Direct correlations between afatinib treatment and PGx profiles were not observed.
    CONCLUSIONS: An afatinib starting dose of 30 mg/day could be an effective and safe treatment option for elderly patients.
    Keywords:  EGFR mutation; Elderly patients; Low-dose afatinib; Pharmacogenomics; Pharmacokinetics
    DOI:  https://doi.org/10.1016/j.ejca.2021.10.024
  121. Ann Hepatobiliary Pancreat Surg. 2021 Nov 30. 25(4): 477-484
    Preoperative Prognostic Nutritional Index is a significant predictive factor for posthepatectomy bile leakage.
    Atsushi Nanashima, Masahide Hiyoshi, Naoya Imamura, Koichi Yano, Takeomi Hamada, Kengo Kai, Takahiro Nishida, Yukako Uchise, Risa Sakamoto, Mayu Inomata.
      Backgrounds/Aims: It is known that preoperative nutritional status can influence patient outcomes after hepatectomy. Prognostic Nutritional Index (PNI) is a useful parameter to reflect patient outcomes undergoing gastro-intestinal surgery. The aim of this study was to retrospectively evaluate relationships of nutritional parameters, demographics, and surgical records with postoperative outcomes in a cohort study.Methods: Curative hepatectomy was performed for 182 patients at the University of Miyazaki between 2015 and 2018. Each preoperative level of albumin, prealbumin, lymphocyte, total cholesterol, or the comprehensively calculated Onodera's PNI was examined as a nutritional parameter.
    Results: The mean PNI was 39.6 ± 5.1, with PNI below 40 observed in 91 (50.0%) patients. Nutritional parameters were not different among patients with various liver diseases. Serum albumin or prealbumin level was significantly correlated with each hepatic parameter (p < 0.01). Prealbumin and total cholesterol levels were significantly correlated with postoperative prothrombin activity (p < 0.05). Albumin or prealbumin levels and PNI were significantly lower in patients with posthepatectomy complications, particularly bile leakage in comparison those without such complications (p < 0.05). Multiple logistic analysis showed that albumin level was an independent risk factor for complications after hepatectomy (risk ratio [RR]: 1.33) and that lymphocyte count was an independent risk factor for bile leakage (RR: 1.28) (p < 0.05). The cut-off level of albumin was approximately 3.8 mg/dL and that of lymphocyte count was 1,320/mm3.
    Conclusions: Preoperative PNI reflected perioperative liver functional status. It was a predictive parameter for postoperative complications, particularly biliary leakage.
    Keywords:  Hepatectomy; Postoperative complications; Postoperative nutritional parameters; Prognostic Nutritional Index
    DOI:  https://doi.org/10.14701/ahbps.2021.25.4.477
  122. Thorac Cancer. 2021 Nov 28.
    Comprehensive analysis of PD-L1 in non-small cell lung cancer with emphasis on survival benefit, impact of driver mutation and histological types, and archival tissue.
    Chin-Chou Wang, Kuo-Tung Huang, Huang-Chih Chang, Chia-Cheng Tseng, Chien-Hao Lai, Jui Lan, Ting-Ting Liu, Chao-Cheng Huang, Meng-Chih Lin.
      BACKGROUND: The aim of the study was to assess programmed death-ligand-1 (PD-L1) expression in different histological types and gene mutation status of patients with non-small cell lung cancer (NSCLC).METHODS: A total of 4062 pathology-confirmed lung cancer patients were retrospectively screened at Kaohsiung Chang Gung Memorial Hospital from November 2010 to June 2017. There were 699 NSCLC patients with confirmed PD-L1 expression level retrospectively enrolled for analysis.
    RESULTS: There was a trend of higher PD-L1 expression in squamous cell carcinoma and adenosquamous cell carcinoma than in adenocarcinoma (p = 063). Significant higher PD-L1 expression in EGFR wild-type was noted (p < 0.001). No significant differences in PD-L1 expression were found between ALK wild- and mutant types, but there seem was a trend of high PD-L1 level noted in ALK mutation patients (p = 0.069). In EGFR mutation patients, a higher time to treatment failure (TTF) duration was observed in no PD-L1 expression (p = 0.011). Longer tumor tissue storage time correlated with lower PD-L1 expression in lung cancer (p < 0.001 for linear trend).
    CONCLUSIONS: There were a trend or significant differences in PD-L1 expression between different histological types in NSCLC, different EGFR and ALK status, and different tumor tissue storage time. A higher survival benefit was observed in no PD-L1 expression than with PD-L1 expression in adenocarcinoma, EGFR and ALK mutation patients. We recommend that PD-L1 assay should be performed as early as possible if tissue is available.
    Keywords:  22C3 IHC assay; ALK; EGFR; PD-L1 expression; non-small cell lung cancer (NSCLC)
    DOI:  https://doi.org/10.1111/1759-7714.14216
  123. J Funct Biomater. 2021 Nov 11. pii: 60. [Epub ahead of print]12(4):
    Evaluation of Clinical Performance of TiNi-Based Implants Used in Chest Wall Repair after Resection for Malignant Tumors.
    Evgeniy Topolnitskiy, Timofey Chekalkin, Ekaterina Marchenko, Yuri Yasenchuk, Seung-Baik Kang, Ji-Hoon Kang, Aleksei Obrosov.
      In this study, we assessed the outcomes after surgical treatment of thoracic post-excision defects in 15 patients, using TiNi knitted surgical meshes and customized artificial TiNi-based ribs.METHODS: Eight patients were diagnosed with advanced non-small cell lung cancer (NSCLC) invading the chest wall, of which five patients were T3N0M0, two were T3N1M0, and one was T3N2M0. Squamous cell carcinoma was identified in three of these patients and adenocarcinoma in five. In two cases, chest wall resection and repair were performed for metastases of kidney cancer after radical nephrectomy. Three-dimensional CT reconstruction and X-ray scans were used to plan the surgery and customize the reinforcing TiNi-based implants. All patients received TiNi-based devices and were prospectively followed for a few years.
    RESULTS: So far, there have been no lethal outcomes, and all implanted devices were consistent in follow-up examinations. Immediate complications were noted in three cases (ejection of air through the pleural drains, paroxysm of atrial fibrillation, and pleuritis), which were conservatively managed. In the long term, no complications, aftereffects, or instability of the thoracic cage were observed.
    CONCLUSION: TiNi-based devices used for extensive thoracic lesion repair in this context are promising and reliable biomaterials that demonstrate good functional, clinical, and cosmetic outcomes.
    Keywords:  TiNi artificial rib; TiNi mesh implant; chest wall reconstruction; non-small cell lung cancer (NSCLC); thoracic lesion
    DOI:  https://doi.org/10.3390/jfb12040060
  124. ACS Sens. 2021 Dec 02.
    Simultaneous Evaluations of pH and Enzyme Activity with a CEST MRI Contrast Agent.
    Chathuri J Kombala, Sanjaya D Lokugama, Aikaterini Kotrotsou, Tianzhe Li, Alyssa C Pollard, Mark D Pagel.
      The extracellular tumor microenvironment of many solid tumors has high acidosis and high protease activity. Simultaneously assessing both characteristics may improve diagnostic evaluations of aggressive tumors and the effects of anticancer treatments. Noninvasive imaging methods have previously been developed that measure extracellular pH or can detect enzyme activity using chemical exchange saturation transfer (CEST) magnetic resonance imaging (MRI). Herein, we developed a single-hybrid CEST agent that can simultaneously measure pH and evaluate protease activity using a combination of dual-power acidoCEST MRI and catalyCEST MRI. Our agent showed CEST signals at 9.2 ppm from a salicylic acid moiety and at 5.0 ppm from an aryl amide. The CEST signal at 9.2 ppm could be measured after selective saturation was applied at 1 and 4 μT, and these measurements could be used with a ratiometric analysis to determine pH. The CEST signal at 5.0 ppm from the aryl amide disappeared after the agent was treated with cathepsin B, while the CEST signal at 9.2 ppm remained, indicating that the agent could detect protease activity through the amide bond cleavage. Michaelis-Menten kinetics studies with catalyCEST MRI demonstrated that the binding affinity (as shown with the Michaelis constant KM), the catalytic turnover rate (kcat), and catalytic efficiency (kcat/KM) were each higher for cathepsin B at lower pH. The kcat rates measured with catalyCEST MRI were lower than the comparable rates measured with liquid chromatography-mass spectrometry (LC-MS), which reflected a limitation of inherently noisy and relatively insensitive CEST MRI analyses. Although this level of precision limited catalyCEST MRI to semiquantitative evaluations, these semiquantitative assessments of high and low protease activity still had value by demonstrating that high acidosis and high protease activity can be used as synergistic, multiparametric biomarkers.
    Keywords:  CEST; Michaelis−Menten kinetics; enzyme activity; magnetic resonance imaging; pH
    DOI:  https://doi.org/10.1021/acssensors.1c02408
  125. Tuberc Respir Dis (Seoul). 2021 Dec 01.
    Afatinib mediates autophagic degradation of ORAI1, STIM1, and SERCA2, which inhibits proliferation of non-small cell lung cancer cells.
    Mi Seong Kim, So Hui Kim, Sei-Hoon Yang, Min Seuk Kim.
      Background: The expression of calcium signaling pathway molecules is altered in various carcinomas, which are related to the proliferation and altered characteristics of cancer cells. However, changes in calcium signaling in anti-cancer drug resistant cells (bearing a T790M mutation in EGFR) remain unclear.Methods: Afatinib-mediated changes in the level of SOCE-related proteins and intracellular Ca2+ level in non-small cell lung cancer cells with T790M mutation in the EGFR gene were analyzed using western blot and ratiometric assays, respectively. Afatinib-mediated autophagic flux was evaluated by measuring cleavage of LC3B-II. Flow cytometry and cell proliferation assays were conducted to assess cell apoptosis and proliferation.
    Results: The levels of SOCE-mediating proteins (ORAI1, STIM1, and SERCA2) decreased after afatinib treatment in non-small cell lung cancer cells, whereas the levels of SOCE-related proteins did not reduce in gefitinib-resistant non-small cell lung cancer cells (PC-9/GR; bearing a T790M mutation in EGFR). Notably, the expression level of SOCE-related proteins in PC-9/GR cells was reduced similarly responding to afatinib in the absence of extracellular Ca2+. Moreover, extracellular Ca2+ influx through the SOCE was significantly reduced in PC-9 cells pre-treated with afatinib than in the control group. Additionally, afatinib was found to decrease the level of SOCE-related proteins through autophagic degradation, and the proliferation of PC-9GR cells was significantly inhibited by a lack of extracellular Ca2+.
    Conclusion: Extracellular Ca2+ plays important roles in afatinib-mediated autophagic degradation of SOCE-related proteins in cells with T790 mutation in the EGFR gene and extracellular Ca2+ is essential for determining anti-cancer drug efficacy.
    Keywords:  Afatinib; Calcium channels; Non-small cell lung; autophagy
    DOI:  https://doi.org/10.4046/trd.2021.0095
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