bims-mascan Biomed News
on Mass spectrometry in cancer research
Issue of 2021–05–23
twenty-two papers selected by
Giovanny Rodríguez Blanco, University of Edinburgh
  1. Measuring MYC-Mediated Metabolism in Tumorigenesis.
  2. MYCN mediates TFRC-dependent ferroptosis and reveals vulnerabilities in neuroblastoma.
  3. Simultaneous measurement of 13 circulating vitamin D3 and D2 mono and dihydroxy metabolites using liquid chromatography mass spectrometry.
  4. Multiblock metabolomics: An approach to elucidate whole-body metabolism with multiblock principal component analysis.
  5. Glutamine Metabolism in Cancer.
  6. Preparation of Human Tissues Embedded in Optimal Cutting Temperature Compound for Mass Spectrometry Analysis.
  7. The Heterogeneity of Lipid Metabolism in Cancer.
  8. SUCLA2-coupled regulation of GLS succinylation and activity counteracts oxidative stress in tumor cells.
  9. A New Workflow for Drug Metabolite Profiling by Utilizing Advanced Tribrid Mass Spectrometry and Data-Processing Techniques.
  10. Targeting Metabolic Cross Talk Between Cancer Cells and Cancer-Associated Fibroblasts.
  11. Revealing Fatty Acid Heterogeneity in Staphylococcal Lipids with Isotope Labeling and RPLC-IM-MS.
  12. Characterization of brain-derived extracellular vesicle lipids in Alzheimer's disease.
  13. Quantifying Precision Loss in Targeted Metabolomics Based on Mass Spectrometry and Nonmatching Internal Standards.
  14. Single-cell lipidomics with high structural specificity by mass spectrometry.
  15. The colorectal cancer lipidome - identification of a robust tumor-specific lipid species signature.
  16. Ferritinophagy and ferroptosis in the management of metabolic diseases.
  17. Human cancer xenografts in immunocompromised mice provide an advanced genuine tumor model for research and drug development-A revisit of murine models for human cancers.
  18. SMYD2 suppresses p53 activity to promote glucose metabolism in cervical cancer.
  19. Global profiling of gut microbiota-associated lipid metabolites in antibiotic-treated mice by LC-MS/MS-based analyses.
  20. Ferroptosis in Ovarian Cancer: A Novel Therapeutic Strategy.
  21. Identifying Protein-(Hydroxy)Methylated DNA Interactions Using Quantitative Interaction Proteomics.
  22. Estimating Global Methylation and Erasure Using Low-Coverage Whole-Genome Bisulfite Sequencing (WGBS ).
  1. Methods Mol Biol. 2021 ;2318 231-239
    Measuring MYC-Mediated Metabolism in Tumorigenesis.
    Hsin-Yao Tang, Aaron R Goldman, Xue Zhang, David W Speicher, Chi V Dang.
      The MYC gene regulates normal cell growth and is deregulated in many human cancers, contributing to tumor growth and progression. The MYC transcription factor activates RNA polymerases I, II, and III target genes that are considered housekeeping genes. These target genes are largely involved in ribosome biogenesis, fatty acid, protein and nucleotide synthesis, nutrient influx or metabolic waste efflux, glycolysis, and glutamine metabolism. MYC's function as a driver of cell growth has been revealed through RNA sequencing, genome-wide chromatin immunoprecipitation, proteomics, and importantly metabolomics, which is highlighted in this chapter.
    Keywords:  Cancer metabolism; MYC; Mass spectrometry; Metabolomics; Transcription
    DOI:  https://doi.org/10.1007/978-1-0716-1476-1_11
  2. Cell Death Dis. 2021 May 19. 12(6): 511
    MYCN mediates TFRC-dependent ferroptosis and reveals vulnerabilities in neuroblastoma.
    Yuxiong Lu, Qing Yang, Yubin Su, Yin Ji, Guobang Li, Xianzhi Yang, Liyan Xu, Zhaoliang Lu, Jiajun Dong, Yi Wu, Jin-Xin Bei, Chaoyun Pan, Xiaoqiong Gu, Bo Li.
      MYCN amplification is tightly associated with the poor prognosis of pediatric neuroblastoma (NB). The regulation of NB cell death by MYCN represents an important aspect, as it directly contributes to tumor progression and therapeutic resistance. However, the relationship between MYCN and cell death remains elusive. Ferroptosis is a newly identified cell death mode featured by lipid peroxide accumulation that can be attenuated by GPX4, yet whether and how MYCN regulates ferroptosis are not fully understood. Here, we report that MYCN-amplified NB cells are sensitive to GPX4-targeting ferroptosis inducers. Mechanically, MYCN expression reprograms the cellular iron metabolism by upregulating the expression of TFRC, which encodes transferrin receptor 1 as a key iron transporter on the cell membrane. Further, the increased iron uptake promotes the accumulation of labile iron pool, leading to enhanced lipid peroxide production. Consistently, TFRC overexpression in NB cells also induces selective sensitivity to GPX4 inhibition and ferroptosis. Moreover, we found that MYCN fails to alter the general lipid metabolism and the amount of cystine imported by System Xc(-) for glutathione synthesis, both of which contribute to ferroptosis in alternative contexts. In conclusion, NB cells harboring MYCN amplification are prone to undergo ferroptosis conferred by TFRC upregulation, suggesting that GPX4-targeting ferroptosis inducers or TFRC agonists can be potential strategies in treating MYCN-amplified NB.
    DOI:  https://doi.org/10.1038/s41419-021-03790-w
  3. Clin Chem Lab Med. 2021 May 19.
    Simultaneous measurement of 13 circulating vitamin D3 and D2 mono and dihydroxy metabolites using liquid chromatography mass spectrometry.
    Carl Jenkinson, Reena Desai, Andrzej T Slominski, Robert C Tuckey, Martin Hewison, David J Handelsman.
       OBJECTIVES: Clinical evaluation of vitamin D status is conventionally performed by measuring serum levels of a single vitamin D metabolite, 25-hydroxyvitamin D predominantly by immunoassay methodology. However, this neglects the complex metabolic pathways involved in vitamin D bioactivity, including two canonical forms D3 and D2, bioactive 1,25-dihydroxy metabolites and inactive 24-hydroxy and other metabolites.
    METHODS: Liquid chromatography-tandem mass spectrometry (LC-MS/MS) can measure multiple analytes in a sample during a single run with high sensitivity and reference level specificity. We therefore aimed to develop and validate a LC-MS/MS method to measure simultaneously 13 circulating vitamin D metabolites and apply it to 103 human serum samples.
    RESULTS: The LC-MS/MS method using a Cookson-type derivatization reagent phenyl-1,2,4-triazoline-3,5-dione (PTAD) quantifies 13 vitamin D metabolites, including mono and dihydroxy-metabolites, as well as CYP11A1-derived D3 and D2 metabolites in a single run. The lower limit of quantitation was 12.5 pg/mL for 1,25(OH)2D3 with accuracy verified by analysis of National Institute of Standards and Technology (NIST) 972a standards. Quantification of seven metabolites (25(OH)D3, 25(OH)D2, 3-epi-25(OH)D3, 20(OH)D3, 24,25(OH)2D3, 1,25(OH)2D3 and 1,20S(OH)2D3) was consistently achieved in human serum samples.
    CONCLUSIONS: This profiling method can provide new insight into circulating vitamin D metabolite pathways forming the basis for improved understanding of the role of vitamin D in health and disease.
    Keywords:  liquid chromatography; mass spectrometry; metabolism; vitamin D
    DOI:  https://doi.org/10.1515/cclm-2021-0441
  4. Comput Struct Biotechnol J. 2021 ;19 1956-1965
    Multiblock metabolomics: An approach to elucidate whole-body metabolism with multiblock principal component analysis.
    Kazuhiro Tanabe, Chihiro Hayashi, Tomoko Katahira, Katsuhiko Sasaki, Ko Igami.
      Principal component analysis (PCA) is a useful tool for omics analysis to identify underlying factors and visualize relationships between biomarkers. However, this approach is limited in addressing life complexity and further improvement is required. This study aimed to develop a new approach that combines mass spectrometry-based metabolomics with multiblock PCA to elucidate the whole-body global metabolic network, thereby generating comparable metabolite maps to clarify the metabolic relationships among several organs. To evaluate the newly developed method, Zucker diabetic fatty (ZDF) rats (n = 6) were used as type 2 diabetic models and Sprague Dawley (SD) rats (n = 6) as controls. Metabolites in the heart, kidney, and liver were analyzed by capillary electrophoresis and liquid chromatography mass spectrometry, respectively, and the detected metabolites were analyzed by multiblock PCA. More than 300 metabolites were detected in the heart, kidney, and liver. When the metabolites obtained from the three organs were analyzed with multiblock PCA, the score and loading maps obtained were highly synchronized and their metabolism patterns were visually comparable. A significant finding in this study was the different expression patterns in lipid metabolism among the three organs; notably triacylglycerols with polyunsaturated fatty acids or less unsaturated fatty acids showed specific accumulation patterns depending on the organs.
    Keywords:  AMP, adenosine monophosphate; Biomarkers; CE/MS, capillary electrophoresis mass spectrometry; CV, coefficient of variation; ESI, electrospray ionization; FABP, fatty acid-binding protein; GC/MS, gas chromatography mass spectrometry; LC/MS, liquid chromatography mass spectrometry; Mass spectrometry; Metabolomics; Multiblock PCA; PCA, principal component analysis; PPAR, peroxisome proliferator-activated receptor; QC, quality control; SD, Sprague Dawley; TCA, tricarboxylic acid. CoA, coenzyme A; TG, triacylglycerol; Type 2 Diabetes; UPLC, ultra-performance liquid chromatography; ZDF, Zucker diabetic fatty
    DOI:  https://doi.org/10.1016/j.csbj.2021.04.015
  5. Adv Exp Med Biol. 2021 ;1311 17-38
    Glutamine Metabolism in Cancer.
    Ting Li, Christopher Copeland, Anne Le.
      Metabolism is a fundamental process for all cellular functions. For decades, there has been growing evidence of a relationship between metabolism and malignant cell proliferation. Unlike normal differentiated cells, cancer cells have reprogrammed metabolism in order to fulfill their energy requirements. These cells display crucial modifications in many metabolic pathways, such as glycolysis and glutaminolysis, which include the tricarboxylic acid (TCA) cycle, the electron transport chain (ETC), and the pentose phosphate pathway (PPP) [1]. Since the discovery of the Warburg effect, it has been shown that the metabolism of cancer cells plays a critical role in cancer survival and growth. More recent research suggests that the involvement of glutamine in cancer metabolism is more significant than previously thought. Glutamine, a nonessential amino acid with both amine and amide functional groups, is the most abundant amino acid circulating in the bloodstream [2]. This chapter discusses the characteristic features of glutamine metabolism in cancers and the therapeutic options to target glutamine metabolism for cancer treatment.
    Keywords:  Glutamine addiction; Glutamine metabolism; Targeting amino acid synthesis; Targeting glutamine metabolism; Transaminase upregulation
    DOI:  https://doi.org/10.1007/978-3-030-65768-0_2
  6. J Vis Exp. 2021 Apr 27.
    Preparation of Human Tissues Embedded in Optimal Cutting Temperature Compound for Mass Spectrometry Analysis.
    April E Boyd, Jeremy Allegood, Santiago Lima.
      Sphingolipids are cellular components that have well-established roles in human metabolism and disease. Mass spectrometry can be used to determine whether sphingolipids are altered in a disease and investigate whether sphingolipids can be targeted clinically. However, properly powered prospective studies that acquire tissues directly from the surgical suite can be time consuming, and technically, logistically, and administratively challenging. In contrast, retrospective studies can take advantage of cryopreserved human specimens already available, usually in large numbers, at tissue biorepositories. Other advantages of procuring tissues from biorepositories include access to information associated with the tissue specimens including histology, pathology, and in some instances clinicopathological variables, all of which can be used to examine correlations with lipidomics data. However, technical limitations related to the incompatibility of optimal cutting temperature compound (OCT) used in the cryopreservation and mass spectrometry is a technical barrier for the analysis of lipids. However, we have previously shown that OCT can be easily removed from human biorepository specimens through cycles of washes and centrifugation without altering their sphingolipid content. We have also previously established that sphingolipids in human tissues cryopreserved in OCT are stable for up to 16 years. In this report, we outline the steps and workflow to analyze sphingolipids in human tissue specimens that are embedded in OCT, including washing tissues, weighing tissues for data normalization, the extraction of lipids, preparation of samples for analysis by liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS), mass spectrometry data integration, data normalization, and data analysis.
    DOI:  https://doi.org/10.3791/62552
  7. Adv Exp Med Biol. 2021 ;1311 39-56
    The Heterogeneity of Lipid Metabolism in Cancer.
    Joshua K Park, Nathan J Coffey, Aaron Limoges, Anne Le.
      The study of cancer cell metabolism has traditionally focused on glycolysis and glutaminolysis. However, lipidomic technologies have matured considerably over the last decade and broadened our understanding of how lipid metabolism is relevant to cancer biology [1-3]. Studies now suggest that the reprogramming of cellular lipid metabolism contributes directly to malignant transformation and progression [4, 5]. For example, de novo lipid synthesis can supply proliferating tumor cells with phospholipid components that comprise the plasma and organelle membranes of new daughter cells [6, 7]. Moreover, the upregulation of mitochondrial β-oxidation can support tumor cell energetics and redox homeostasis [8], while lipid-derived messengers can regulate major signaling pathways or coordinate immunosuppressive mechanisms [9-11]. Lipid metabolism has, therefore, become implicated in a variety of oncogenic processes, including metastatic colonization, drug resistance, and cell differentiation [10, 12-16]. However, whether we can safely and effectively modulate the underlying mechanisms of lipid metabolism for cancer therapy is still an open question.
    Keywords:  Cancer metabolism; Fatty acid oxidation; Fatty acid uptake; Lipid synthesis; Lipidomics; Metastasis; Tumor heterogeneity
    DOI:  https://doi.org/10.1007/978-3-030-65768-0_3
  8. Mol Cell. 2021 May 08. pii: S1097-2765(21)00269-0. [Epub ahead of print]
    SUCLA2-coupled regulation of GLS succinylation and activity counteracts oxidative stress in tumor cells.
    Yingying Tong, Dong Guo, Shu-Hai Lin, Jiazhen Liang, Dianqiang Yang, Chunmin Ma, Fei Shao, Min Li, Qiujing Yu, Yuhui Jiang, Lei Li, Jing Fang, Rilei Yu, Zhimin Lu.
      Glutaminase regulates glutaminolysis to promote cancer cell proliferation. However, the mechanism underlying glutaminase activity regulation is largely unknown. Here, we demonstrate that kidney-type glutaminase (GLS) is highly expressed in human pancreatic ductal adenocarcinoma (PDAC) specimens with correspondingly upregulated glutamine dependence for PDAC cell proliferation. Upon oxidative stress, the succinyl-coenzyme A (CoA) synthetase ADP-forming subunit β (SUCLA2) phosphorylated by p38 mitogen-activated protein kinase (MAPK) at S79 dissociates from GLS, resulting in enhanced GLS K311 succinylation, oligomerization, and activity. Activated GLS increases glutaminolysis and the production of nicotinamide adenine dinucleotide phosphate (NADPH) and glutathione, thereby counteracting oxidative stress and promoting tumor cell survival and tumor growth in mice. In addition, the levels of SUCLA2 pS79 and GLS K311 succinylation, which were mutually correlated, were positively associated with advanced stages of PDAC and poor prognosis for patients. Our findings reveal critical regulation of GLS by SUCLA2-coupled GLS succinylation regulation and underscore the regulatory role of metabolites in glutaminolysis and PDAC development.
    Keywords:  GLS; GSH; NADPH; SUCLA2; glutaminolysis; p38; phosphorylation; succinyl-CoA; succinylation; tumorigenesis
    DOI:  https://doi.org/10.1016/j.molcel.2021.04.002
  9. J Am Soc Mass Spectrom. 2021 May 17.
    A New Workflow for Drug Metabolite Profiling by Utilizing Advanced Tribrid Mass Spectrometry and Data-Processing Techniques.
    Qian Ruan, Kate Comstock.
      Drug metabolite profiling utilizes liquid chromatography with tandem mass spectrometry (LC/MS/MS) to acquire ample information for metabolite identification and structural elucidation. However, there are still challenges in detecting and characterizing all potential metabolites that can be masked by a high biological background, especially the unknown and uncommon ones. In this work, a novel metabolite profiling workflow was established on a platform using a state-of-the-art tribrid high-resolution mass spectrometry (HRMS) system. Primarily, an instrumental method was developed based on the novel design of the tribrid system that facilitates in-depth MSn scans with two fragmentation devices. Additionally, different advanced data acquisition techniques were assessed and compared, and automatic background exclusion and deep-scan approaches were adopted to promote assay efficiency and metabolite coverage. Finally, different data-analysis techniques were explored to fully extract metabolite data from the information-rich MS/MS data sets. Overall, a workflow combining tribrid mass spectrometry and advanced acquisition methodology has been developed for metabolite characterization in drug discovery and development. It maximizes the tribrid HRMS platform's utility and enhances the coverage, efficiency, quality, and speed of metabolite profiling assays.
    DOI:  https://doi.org/10.1021/jasms.0c00436
  10. Adv Exp Med Biol. 2021 ;1311 205-214
    Targeting Metabolic Cross Talk Between Cancer Cells and Cancer-Associated Fibroblasts.
    Jin G Jung, Anne Le.
      Although cancer has classically been regarded as a genetic disease of uncontrolled cell growth, the importance of the tumor microenvironment (TME) [1, 2] is continuously emphasized by the accumulating evidence that cancer growth is not simply dependent on the cancer cells themselves [3, 4] but also dependent on angiogenesis [5-8], inflammation [9, 10], and the supporting roles of cancer-associated fibroblasts (CAFs) [11-13]. After the discovery that CAFs are able to remodel the tumor matrix within the TME and provide the nutrients and chemicals to promote cancer cell growth [14], many studies have aimed to uncover the cross talk between cancer cells and CAFs. Moreover, a new paradigm in cancer metabolism shows how cancer cells act like "metabolic parasites" to take up the high-energy metabolites, such as lactate, ketone bodies, free fatty acids, and glutamine from supporting cells, including CAFs and cancer-associated adipocytes (CAAs) [15, 16]. This chapter provides an overview of the metabolic coupling between CAFs and cancer cells to further define the therapeutic options to disrupt the CAF-cancer cell interactions.
    Keywords:  Cancer therapy; Cancer-associated adipocytes; Cancer-associated fibroblasts; Metabolism; Metabolites; Tumor microenvironment
    DOI:  https://doi.org/10.1007/978-3-030-65768-0_15
  11. J Am Soc Mass Spectrom. 2021 May 20.
    Revealing Fatty Acid Heterogeneity in Staphylococcal Lipids with Isotope Labeling and RPLC-IM-MS.
    Christian Freeman, Hannah M Hynds, Jana M Carpenter, Keerthi Appala, Kingsley Bimpeh, Shannon Barbarek, Craig Gatto, Brian J Wilkinson, Kelly M Hines.
      Up to 80% of the fatty acids in Staphylococcus aureus membrane lipids are branched, rather than straight-chain, fatty acids. The branched fatty acids (BCFAs) may have either an even or odd number of carbons, and the branch position may be at the penultimate carbon (iso) or the antepenultimate (anteiso) carbon of the tail. This results in two sets of isomeric fatty acid species with the same number of carbons that cannot be resolved by mass spectrometry. The isomer/isobar challenge is further complicated when the mixture of BCFAs and straight-chain fatty acids (SCFAs) are esterified into diacylated lipids such as the phosphatidylglycerol (PG) species of the S. aureus membrane. No conventional chromatographic method has been able to resolve diacylated lipids containing mixtures of SCFAs, anteiso-odd, iso-odd, and iso-even BCFAs. A major hurdle to method development in this area is the lack of relevant analytical standards for lipids containing BCFA isomers. The diversity of the S. aureus lipidome and its naturally high levels of BCFAs present an opportunity to explore the potential of resolving diacylated lipids containing BCFAs and SFCAs. Using our knowledge of lipid and fatty acid biosynthesis in S. aureus, we have used a stable-isotope-labeling strategy to develop and validate a 30 min C18 reversed-phase liquid chromatography method combined with traveling-wave ion mobility-mass spectrometry to provide resolution of diacylated lipids based on the number of BCFAs that they contain.
    Keywords:  Staphylococcus aureus; branched fatty acids; isomers; isotope labeling; lipids; reversed-phase liquid chromatography
    DOI:  https://doi.org/10.1021/jasms.1c00092
  12. J Extracell Vesicles. 2021 May;10(7): e12089
    Characterization of brain-derived extracellular vesicle lipids in Alzheimer's disease.
    Huaqi Su, Yepy H Rustam, Colin L Masters, Enes Makalic, Catriona A McLean, Andrew F Hill, Kevin J Barnham, Gavin E Reid, Laura J Vella.
      Lipid dyshomeostasis is associated with the most common form of dementia, Alzheimer's disease (AD). Substantial progress has been made in identifying positron emission tomography and cerebrospinal fluid biomarkers for AD, but they have limited use as front-line diagnostic tools. Extracellular vesicles (EVs) are released by all cells and contain a subset of their parental cell composition, including lipids. EVs are released from the brain into the periphery, providing a potential source of tissue and disease specific lipid biomarkers. However, the EV lipidome of the central nervous system is currently unknown and the potential of brain-derived EVs (BDEVs) to inform on lipid dyshomeostasis in AD remains unclear. The aim of this study was to reveal the lipid composition of BDEVs in human frontal cortex, and to determine whether BDEVs have an altered lipid profile in AD. Using semi-quantitative mass spectrometry, we describe the BDEV lipidome, covering four lipid categories, 17 lipid classes and 692 lipid molecules. BDEVs were enriched in glycerophosphoserine (PS) lipids, a characteristic of small EVs. Here we further report that BDEVs are enriched in ether-containing PS lipids, a finding that further establishes ether lipids as a feature of EVs. BDEVs in the AD frontal cortex offered improved detection of dysregulated lipids in AD over global lipid profiling of this brain region.  AD BDEVs had significantly altered glycerophospholipid and sphingolipid levels, specifically increased plasmalogen glycerophosphoethanolamine and decreased polyunsaturated fatty acyl containing lipids, and altered amide-linked acyl chain content in sphingomyelin and ceramide lipids relative to CTL. The most prominent alteration was a two-fold decrease in lipid species containing anti-inflammatory/pro-resolving docosahexaenoic acid. The in-depth lipidome analysis provided in this study highlights the advantage of EVs over more complex tissues for improved detection of dysregulated lipids that may serve as potential biomarkers in the periphery.
    Keywords:  Alzheimer's disease; brain; exosomes; extracellular vesicles; frontal cortex; glycerophospholipids; lipid biomarkers; lipidome; polyunsaturated fatty acids; sphingolipids; tissue
    DOI:  https://doi.org/10.1002/jev2.12089
  13. Anal Chem. 2021 May 20.
    Quantifying Precision Loss in Targeted Metabolomics Based on Mass Spectrometry and Nonmatching Internal Standards.
    Arve Ulvik, Adrian McCann, Øivind Midttun, Klaus Meyer, Keith M Godfrey, Per M Ueland.
      In mass spectrometry, reliable quantification requires correction for variations in ionization efficiency between samples. The preferred method is the addition of a stable isotope-labeled internal standard (SIL-IS). In targeted metabolomics, a dedicated SIL-IS for each metabolite of interest may not always be realized due to high cost or limited availability. We recently completed the analysis of more than 70 biomarkers, each with a matching SIL-IS, across four mass spectrometry-based platforms (one GC-MS/MS and three LC-MS/MS). Using data from calibrator and quality control samples added to 60 96-well trays (analytical runs), we calculated analytical precision (CV) retrospectively. The use of integrated peak areas for all metabolites and internal standards allowed us to calculate precision for all matching analyte (A)/SIL-IS (IS) pairs as well as for all nonmatching A/IS pairs within each platform (total n = 1442). The median between-run precision for matching A/IS across the four platforms was 2.7-5.9%. The median CV for nonmatching A/IS (corresponding to pairing analytes with a non-SIL-IS) was 2.9-10.7 percentage points higher. Across all platforms, CVs for nonmatching A/IS increased with increasing difference in retention time (Spearman's rho of 0.17-0.93). The CV difference for nonmatching vs matching A/IS was often, but not always, smaller when analytes and internal standards were close structural analogs.
    DOI:  https://doi.org/10.1021/acs.analchem.1c00119
  14. Nat Commun. 2021 05 17. 12(1): 2869
    Single-cell lipidomics with high structural specificity by mass spectrometry.
    Zishuai Li, Simin Cheng, Qiaohong Lin, Wenbo Cao, Jing Yang, Minmin Zhang, Aijun Shen, Wenpeng Zhang, Yu Xia, Xiaoxiao Ma, Zheng Ouyang.
      Single-cell analysis is critical to revealing cell-to-cell heterogeneity that would otherwise be lost in ensemble analysis. Detailed lipidome characterization for single cells is still far from mature, especially when considering the highly complex structural diversity of lipids and the limited sample amounts available from a single cell. We report the development of a general strategy enabling single-cell lipidomic analysis with high structural specificity. Cell fixation is applied to retain lipids in the cell during batch treatments prior to single-cell analysis. In addition to tandem mass spectrometry analysis revealing the class and fatty acyl-chain for lipids, batch photochemical derivatization and single-cell droplet treatment are performed to identify the C=C locations and sn-positions of lipids, respectively. Electro-migration combined with droplet-assisted electrospray ionization enables single-cell mass spectrometry analysis with easy operation but high efficiency in sample usage. Four subtypes of human breast cancer cells are correctly classified through quantitative analysis of lipid C=C location or sn-position isomers in ~160 cells. Most importantly, the single-cell deep lipidomics strategy successfully discriminates gefitinib-resistant cells from a population of wild-type human lung cancer cells (HCC827), highlighting its unique capability to promote precision medicine.
    DOI:  https://doi.org/10.1038/s41467-021-23161-5
  15. Gastroenterology. 2021 May 14. pii: S0016-5085(21)02979-6. [Epub ahead of print]
    The colorectal cancer lipidome - identification of a robust tumor-specific lipid species signature.
    Josef Ecker, Elisa Benedetti, Alida S D Kindt, Marcus Höring, Markus Perl, Andrea Christel Machmüller, Anna Sichler, Johannes Plagge, Yuting Wang, Sebastian Zeissig, Andrej Shevchenko, Ralph Burkhardt, Jan Krumsiek, Gerhard Liebisch, Klaus-Peter Janssen.
       OBJECTIVE: Lipidomic changes were causally linked to metabolic diseases, but the scenario for colorectal cancer (CRC) is less clear. We investigated the CRC lipidome for putative tumour-specific alterations through analysis of three independent retrospective patient cohorts from two clinical centers, to derive a clinically useful signature.
    DESIGN: Quantitative comprehensive lipidomic analysis was performed by direct infusion electrospray ionization coupled to tandem mass spectrometry (ESI-MS/MS) and high-resolution mass spectrometry (HR-MS) on matched non-diseased mucosa and tumor tissue in a discovery cohort (n=106). Results were validated in two independent cohorts (n=28, and n=20), associated with genomic and clinical data, and lipidomic data from a genetic mouse tumor model (Apc1638N).
    RESULTS: Significant differences were found between tumor and normal tissue for glycero-, glycerophospho- and sphingolipids in the discovery cohort. Comparison to the validation collectives unveiled that glycerophospholipids showed high interpatient variation and were strongly affected by preanalytical conditions, whereas glycero- and sphingolipids appeared more robust. Signatures of sphingomyelin (SM) and triacylglycerol (TG) species significantly differentiated cancerous from non-diseased tissue in both validation studies. Moreover, lipogenic enzymes were significantly upregulated in CRC, and FASN gene expression was prognostically detrimental. The TG profile was significantly associated with post-operative disease-free survival and lymphovascular invasion, and was essentially conserved in murine digestive cancer, but not associated with microsatellite status, KRAS or BRAF mutations, or T-cell infiltration.
    CONCLUSION: Analysis of the CRC lipidome revealed a robust TG-species signature with prognostic potential. A better understanding of the cancer-associated glycerolipid and sphingolipid metabolism may lead to novel therapeutic strategies.
    Keywords:  Biomarker; mass spectrometry; signature; sphingomyelin; triacylglycerol
    DOI:  https://doi.org/10.1053/j.gastro.2021.05.009
  16. Trends Endocrinol Metab. 2021 May 15. pii: S1043-2760(21)00095-3. [Epub ahead of print]
    Ferritinophagy and ferroptosis in the management of metabolic diseases.
    Amir Ajoolabady, Hamid Aslkhodapasandhokmabad, Peter Libby, Jaakko Tuomilehto, Gregory Y H Lip, Josef M Penninger, Des R Richardson, Daolin Tang, Hao Zhou, Shuyi Wang, Daniel J Klionsky, Guido Kroemer, Jun Ren.
      Ferroptosis is a form of regulated cell death modality associated with disturbed iron-homeostasis and unrestricted lipid peroxidation. Ample evidence has depicted an essential role for ferroptosis as either the cause or consequence for human diseases, denoting the likely therapeutic promises for targeting ferroptosis in the preservation of human health. Ferritinophagy, a selective form of autophagy, contributes to the initiation of ferroptosis through degradation of ferritin, which triggers labile iron overload (IO), lipid peroxidation, membrane damage, and cell death. In this review, we will delineate the role of ferritinophagy in ferroptosis, and its underlying regulatory mechanisms, to unveil the therapeutic value of ferritinophagy as a target in the combat of ferroptosis to manage metabolic diseases.
    Keywords:  ferritinophagy; ferroptosis; iron overload; lipid peroxidation; metabolic diseases
    DOI:  https://doi.org/10.1016/j.tem.2021.04.010
  17. Biochim Biophys Acta Gen Subj. 2021 May 12. pii: S0304-4165(21)00087-8. [Epub ahead of print]1865(8): 129929
    Human cancer xenografts in immunocompromised mice provide an advanced genuine tumor model for research and drug development-A revisit of murine models for human cancers.
    Quan-En Yang.
      Molecular and cell biology studies have proven that human cancers are an enormously heterogenous disease, even if they originate from the same organ and tissue with identical morphological characteristics. Cancer cells in tumors from different individuals exhibit somewhat different characteristics on multiple levels, such as with respect to 1) their genetic polymorphism; 2) epigenetic mechanisms; 3) group gene activation/inactivation; 4) cell metabolism behavior; 5) aberrant incomplete terminal differentiation; 6) proliferative potential; and 7) hierarchical structure. These multiple parameters and their different combinations determine the biological characteristics of the cancer cells and their malignant/metastatic manifestations. With progress in medical research, numerous unique vulnerable targets of cancer cells have been identified from different tumors. Modern anti-cancer drug development focuses on target-based cancer cell inhibition and elimination have greatly improved the outcome of patients with some specific cancers. The murine model of human cancer has proven to be an essential procedure for the evaluation of drug efficacy in mammalian and a key link in transferring anti-cancer drug from laboratory to clinics. As classical murine cancer xenograft models with different human cancer cell lines display limited value for personalized precision medicine, creating a complete human xenograft cancer bank with all levels of abnormalities in mice has become desperately needed. This article is a review of the pros and cons of different human x murine cancer models and an attempt to find a more suitable model for the study and discovery of new anti-cancer drugs and different combination therapies in this small animal model.
    DOI:  https://doi.org/10.1016/j.bbagen.2021.129929
  18. Exp Cell Res. 2021 May 17. pii: S0014-4827(21)00181-6. [Epub ahead of print] 112649
    SMYD2 suppresses p53 activity to promote glucose metabolism in cervical cancer.
    Ying Wang, Ge Jin, Yunfeng Guo, Yuan Cao, Shuhuai Niu, Xiaomei Fan, Jun Zhang.
      Reprogrammed energy metabolism, especially the Warburg effect, is emerged as a hallmark of cancer. The protein lysine methyltransferase SMYD2 functions as an oncogene and is implicated in various malignant phenotypes of human cancers. However, the role of SMYD2 in tumor metabolism is still largely unknown. Here, we report that SMYD2 is highly expressed in human cervical cancer and its aberrant expression is linked to a poor prognosis. Bioinformatic analysis revealed a novel link between SMYD2 expression and aerobic glycolysis. Through loss-of-function experiments, we demonstrated that SMYD2 knockdown or inhibition induced a metabolic shift from aerobic glycolysis to oxidative phosphorylation, as evidenced by glucose uptake, lactate production, extracellular acidification, and the oxygen consumption rate. In contrast, SMYD2 overexpression promoted glycolytic metabolism in cervical cancer cells. Moreover, SMYD2 was required for tumor growth in cervical cancer and this oncogenic activity was largely glycolysis-dependent. Mechanistically, SMYD2 altered the methylation status of p53 and inhibited its transcriptional activity. Genetic silencing of p53 largely abrogated the effects of SMYD2 in promoting aerobic glycolysis. Taken together, our findings reveal a novel function of SMYD2 in regulating the Warburg effect in cervical cancer.
    Keywords:  Cervical cancer; Oxidative phosphorylation; SMYD2; TP53; Warburg effect
    DOI:  https://doi.org/10.1016/j.yexcr.2021.112649
  19. STAR Protoc. 2021 Jun 18. 2(2): 100492
    Global profiling of gut microbiota-associated lipid metabolites in antibiotic-treated mice by LC-MS/MS-based analyses.
    Nobuyuki Okahashi, Masahiro Ueda, Shu Yasuda, Hiroshi Tsugawa, Makoto Arita.
      We describe a protocol for identifying bacteria-derived lipid metabolites produced in the guts using antibiotic-treated mice, liquid chromatography tandem mass spectrometry-based lipidomics, and feature-based molecular spectrum networking (FBMN). Untargeted lipidomics using the MS-DIAL 4 program provides information on known and unknown complex lipid molecules. The FBMN technique clusters similar MS2 spectra, facilitating the identification of bacterial lipids. Targeted analysis was used as a complementary method to cover oxylipins. Here, we provide details for targeted and untargeted analyses. For complete details on the use and execution of this protocol, please refer to Yasuda et al. (2020).
    Keywords:  Immunology; Mass Spectrometry; Metabolomics; Microbiology
    DOI:  https://doi.org/10.1016/j.xpro.2021.100492
  20. Front Oncol. 2021 ;11 665945
    Ferroptosis in Ovarian Cancer: A Novel Therapeutic Strategy.
    Lanyu Li, Cheng Qiu, Min Hou, Xinyu Wang, Changzhen Huang, Jialin Zou, Tianyi Liu, Jinfeng Qu.
      Ovarian cancer (OVCA) is one of the most lethal malignancies with a five-year relative survival below 50% by virtue of its high recurrence rate and inadequate early detection methods. For OVCA patients, modern approaches include debulking surgery, chemotherapies, angiogenesis inhibitors, poly ADP-ribose polymerase (PARP) inhibitors, and immunotherapies depending on the histological type and staging of the tumor. However, in most cases, simple standard treatment is not satisfactory. Thus, a more effective way of treatment is needed. Ferroptosis is a newly recognized type of regulated cell death marked by lipid peroxidation, iron accumulation and glutathione deprivation, having a connection with a variety of disorders and showing great potential in anti-tumor therapy. Intriguingly, a possible connection between ferroptosis and OVCA is shown on the basis of previously published findings. Furthermore, a growing number of ferroptosis protection pathways have been identified during the past few years with increasing ferroptosis regulators being discovered. In this review, we summarized several major pathways involved in ferroptosis and the study foundation of ferroptosis and ovarian cancer, hoping to provide clues regarding OVCA treatment. And some important issues were also raised to point out future research directions.
    Keywords:  ferroptosis; immunotherapy; lipid metabolism; ovarian cancer; reactive oxygen species (ROS)
    DOI:  https://doi.org/10.3389/fonc.2021.665945
  21. Methods Mol Biol. 2021 ;2272 209-224
    Identifying Protein-(Hydroxy)Methylated DNA Interactions Using Quantitative Interaction Proteomics.
    Velin Marita Sequeira, Michiel Vermeulen.
      In recent years, workflows coupling DNA affinity purifications from crude nuclear extracts with quantitative mass spectrometry-based proteomics have enabled comprehensive mapping of protein-DNA interactions in an unbiased manner. Here, we describe a detailed protocol for one such method in which affinity purifications with extracts from cells or tissues of interest are combined with a chemical stable isotope labeling method, dimethyl labeling, to identify specific interaction partners for (hydroxy)methylated and non-methylated DNA sequences of interest.
    Keywords:  Dimethyl labeling; Mass spectrometry; Methylation readers; Protein–methylated DNA interactions; Quantitative proteomics
    DOI:  https://doi.org/10.1007/978-1-0716-1294-1_12
  22. Methods Mol Biol. 2021 ;2272 29-44
    Estimating Global Methylation and Erasure Using Low-Coverage Whole-Genome Bisulfite Sequencing (WGBS ).
    Oscar Ortega-Recalde, Julian R Peat, Donna M Bond, Timothy A Hore.
      Whole-genome bisulfite sequencing (WGBS) is a popular method for characterizing cytosine methylation because it is fully quantitative and has base-pair resolution. While WGBS is prohibitively expensive for experiments involving many samples, low-coverage WGBS can accurately determine global methylation and erasure at similar cost to high-performance liquid chromatography (HPLC) or enzyme-linked immunosorbent assays (ELISA). Moreover, low-coverage WGBS has the capacity to distinguish between methylation in different cytosine contexts (e.g., CG, CHH, and CHG), can tolerate low-input material (<100 cells), and can detect the presence of overrepresented DNA originating from mitochondria or amplified ribosomal DNA. In addition to describing a WGBS library construction and quantitation approach, here we detail computational methods to predict the accuracy of low-coverage WGBS using empirical bootstrap samplers and theoretical estimators similar to those used in election polling. Using examples, we further demonstrate how non-independent sampling of cytosines can alter the precision of error calculation and provide methods to improve this.
    Keywords:  Asymptotic estimator; Bootstrap sampling; DNA methylation; Epigenetics; Low-coverage bisulfite sequencing; Methylation erasure; Post-bisulfite adaptor tagging; Whole-genome bisulfite sequencing
    DOI:  https://doi.org/10.1007/978-1-0716-1294-1_3
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