bims-almceb 2021-03-14 papers

bims-almceb

Biomed News

on Acute Leukemia Metabolism and Cell Biology

Issue of 2021–03–14
nineteen papers selected by
Camila Kehl Dias, Federal University of Rio Grande do Sul

CAR-T cell therapy: practical guide to routine laboratory monitoring.
Blockade of 6-phosphogluconate dehydrogenase generates CD8+ effector T cells with enhanced anti-tumor function.
Clinical and molecular predictors of response and survival following venetoclax therapy in relapsed/refractory AML.
ROS-dependent HIF1α activation under forced lipid catabolism entails glycolysis and mitophagy as mediators of higher proliferation rate in cervical cancer cells.
Signaling pathways in intestinal homeostasis and colorectal cancer: KRAS at centre stage.
Therapeutic and prognostic implications of NOTCH and MAPK signaling in bladder cancer.
Nonphosphorylating Oxidation in Mitochondria and Related Processes.
Strategies to overcome resistance to immune checkpoint blockade in lung cancer.
Mitochondria's Role in the Maintenance of Cancer Stem Cells in Glioblastoma.
Fat induces glucose metabolism in non-transformed liver cells and promotes liver tumorigenesis.
Activation of the MAPK pathway mediates resistance to PI3K inhibitors in chronic lymphocytic leukemia (cll).
Pathway-based classification of glioblastoma uncovers a mitochondrial subtype with therapeutic vulnerabilities.
Proteasome 26S subunit, non-ATPases 1 (PSMD1) and 3 (PSMD3), play an oncogenic role in chronic myeloid leukemia by stabilizing nuclear factor-kappa B.
PSGL-1 Immune Checkpoint Inhibition for CD4+ T Cell Cancer Immunotherapy.
Targeting mitochondrial iron metabolism suppresses tumor growth and metastasis by inducing mitochondrial dysfunction and mitophagy.
Oligomycin A promotes radioresistance in HT29 colorectal cancer cells and its mechanisms.
Oxidative phosphorylation enhances the leukemogenic capacity and resistance to chemotherapy of B cell acute lymphoblastic leukemia.
A Therapeutic Strategy for Preferential Targeting of TET2 Mutant and TET-dioxygenase Deficient Cells in Myeloid Neoplasms.
Balancing ER-Mitochondrial Ca2+ Fluxes in Health and Disease.

Pathology. 2021 Mar 05. pii: S0031-3025(21)00057-X. [Epub ahead of print]

CAR-T cell therapy: practical guide to routine laboratory monitoring.

Adrian G Selim, Adrian Minson, Piers Blombery, Michael Dickinson, Simon J Harrison, Mary Ann Anderson.

  Chimeric antigen receptor (CAR)-T cell therapy is a genetically-modified cellular immunotherapy that has a current established role in the treatment of relapsed/refractory B-cell acute lymphoblastic leukaemia and diffuse large B-cell lymphoma, with emerging utility in a spectrum of other haematological and solid organ malignancies. It is associated with a number of characteristic toxicities, most notably cytokine release syndrome and neurotoxicity, for which laboratory testing can aid in the prediction of severity and in monitoring. Other toxicities, such as cytopenias/marrow hypoplasia, hypogammagloblinaemia and delayed immune reconstitution are recognised and require monitoring due to the implications for infection risk and prophylaxis. The detection or quantitation of circulating CAR-T can be clinically useful, and is achieved through both direct methods, if available, or indirect/surrogate methods. It is important that the laboratory is informed of the CAR-T therapy and target antigen whenever tissue is collected, both for response assessment and investigation of possible relapse, so that the expression of the relevant antigen can be assessed, in order to distinguish antigen-positive and -negative relapses. Finally, the measurement of circulating tumour DNA has an evolving role in the surveillance of malignancy, with evidence of its utility in the post-CAR-T setting, including predicting patients who will inevitably experience frank relapse, potentially allowing for pre-emptive therapy.

Keywords:  CAR-T; molecular; monitoring; toxicity

DOI:  https://doi.org/10.1016/j.pathol.2021.02.002
Cell Rep. 2021 Mar 09. pii: S2211-1247(21)00145-5. [Epub ahead of print]34(10): 108831

Blockade of 6-phosphogluconate dehydrogenase generates CD8+ effector T cells with enhanced anti-tumor function.

Saeed Daneshmandi, Teresa Cassel, Penghui Lin, Richard M Higashi, Gerburg M Wulf, Vassiliki A Boussiotis, Teresa W-M Fan, Pankaj Seth.

  Although T cell expansion depends on glycolysis, T effector cell differentiation requires signaling via the production of reactive oxygen species (ROS). Because the pentose phosphate pathway (PPP) regulates ROS by generating nicotinamide adenine dinucleotide phosphate (NADPH), we examined how PPP blockade affects T cell differentiation and function. Here, we show that genetic ablation or pharmacologic inhibition of the PPP enzyme 6-phosphogluconate dehydrogenase (6PGD) in the oxidative PPP results in the generation of superior CD8+ T effector cells. These cells have gene signatures and immunogenic markers of effector phenotype and show potent anti-tumor functions both in vitro and in vivo. In these cells, metabolic reprogramming occurs along with increased mitochondrial ROS and activated antioxidation machinery to balance ROS production against oxidative damage. Our findings reveal a role of 6PGD as a checkpoint for T cell effector differentiation/survival and evidence for 6PGD as an attractive metabolic target to improve tumor immunotherapy.

Keywords:  6PGD; effector T cells; metabolism; pentose phosphate pathway; reactive oxygen species; tumor immunotherapy

DOI:  https://doi.org/10.1016/j.celrep.2021.108831
Blood Adv. 2021 Mar 09. 5(5): 1552-1564

Clinical and molecular predictors of response and survival following venetoclax therapy in relapsed/refractory AML.

Maximilian Stahl, Kamal Menghrajani, Andriy Derkach, Alexander Chan, Wenbin Xiao, Jacob Glass, Amber C King, Anthony F Daniyan, Christopher Famulare, Bernadette M Cuello, Troy Z Horvat, Omar Abdel-Wahab, Ross L Levine, Aaron D Viny, Eytan M Stein, Sheng F Cai, Mikhail Roshal, Martin S Tallman, Aaron D Goldberg.

Azacitidine + venetoclax, decitabine + venetoclax, and low-dose cytarabine + venetoclax are now standard treatments for newly diagnosed older or unfit patients with acute myeloid leukemia (AML). Although these combinations are also commonly used in relapsed or refractory AML (RR-AML), clinical and molecular predictors of response and survival in RR-AML are incompletely understood. We retrospectively analyzed clinical and molecular characteristics and outcomes for 86 patients with RR-AML who were treated with venetoclax combinations. The complete remission (CR) or CR with incomplete hematologic recovery (CRi) rate was 24%, and the overall response rate was 31% with the inclusion of a morphologic leukemia-free state. Azacitidine + venetoclax resulted in higher response rates compared with low-dose cytarabine + venetoclax (49% vs 15%; P = .008). Median overall survival (OS) was 6.1 months, but it was significantly longer with azacitidine + venetoclax compared with low-dose cytarabine + venetoclax (25 vs 3.9 months; P = .003). This survival advantage of azacitidine + venetoclax over low-dose cytarabine + venetoclax persisted when patients were censored for subsequent allogeneic stem cell transplantation (8.1 vs 3.9 months; P = .035). Mutations in NPM1 were associated with higher response rates, whereas adverse cytogenetics and mutations in TP53, KRAS/NRAS, and SF3B1 were associated with worse OS. Relapse was driven by diverse mechanisms, including acquisition of novel mutations and an increase in cytogenetic complexity. Venetoclax combination therapy is effective in many patients with RR-AML, and pretreatment molecular characteristics may predict outcomes. Trials that evaluate novel agents in combination with venetoclax therapy in patients with RR-AML that have adverse risk genomic features are warranted.

DOI: https://doi.org/10.1182/bloodadvances.2020003734
J Exp Clin Cancer Res. 2021 Mar 11. 40(1): 94

ROS-dependent HIF1α activation under forced lipid catabolism entails glycolysis and mitophagy as mediators of higher proliferation rate in cervical cancer cells.

Serena Castelli, Fabio Ciccarone, Daniela Tavian, Maria Rosa Ciriolo.

   BACKGROUND: In the last decades, the concept of metabolic rewiring as a cancer hallmark has been expanded beyond the "Warburg effect" and the importance of other metabolic routes, including lipid metabolism, has emerged. In cancer, lipids are not only a source of energy but are also required for the formation of membranes building blocks, signaling and post-translational modification of proteins. Since lipid metabolism contributes to the malignancy of cancer cells, it is an attractive target for therapeutic strategies.
METHODS: Over-expression of the adipose triglyceride lipase (ATGL) was used to boost lipid catabolism in cervical cancer cells. The cervical cancer cell line HeLa was employed as the primary experimental model for all subsequent studies. The lipolytic activity of ATGL was mimicked by caproate, a short-chain fatty acid that is efficiently oxidized in mitochondria.
RESULTS: Here, we provide evidence of the association between boosted lipid catabolism and the increased proliferation and migration capability of cervical cancer cells. These pro-tumoral effects were ascribed to the reactive oxygen species (ROS)-mediated induction of hypoxia-inducible factor-1α (HIF1α) triggered by the increased mitochondrial fatty acids (FAs) oxidation. HIF1α activation increases glycolytic flux and lactate production, promoting cell proliferation. At the same time, HIF1α increases protein and mRNA levels of its known target BCL2 and adenovirus E1B 19-kDa-interacting protein 3 (BNIP3), which in turn activates mitophagy as a pro-survival process, as demonstrated by the induction of apoptosis upon inhibition of mitophagy. These effects were mimicked by the short-chain fatty acid caproate, confirming that forcing lipid catabolism results in HIF1α induction.
CONCLUSIONS: Boosting lipid catabolism by ATGL over-expression has a pro-tumor role in cervical cancer cells, dependent on ROS production and HIF1α induction. Together with the bioinformatics evidence of the correlation of ATGL activity with the aggressiveness of cervical cancer cells, our data suggest that ATGL could be a promising prognostic marker for cervical cancer and highlight the need of further investigations on the role of this lipase in cancer cells. This evidence could be exploited to develop new personalized therapy, based on the functionality of the antioxidant equipment of cancer cells, considering that ROS content could affect ATGL role.

Keywords:  ATGL; HIF1α; Lipid catabolism; Mitophagy; Pseudo-hypoxia; ROS

DOI:  https://doi.org/10.1186/s13046-021-01887-w
Cell Commun Signal. 2021 Mar 10. 19(1): 31

Signaling pathways in intestinal homeostasis and colorectal cancer: KRAS at centre stage.

Camille Ternet, Christina Kiel.

  The intestinal epithelium acts as a physical barrier that separates the intestinal microbiota from the host and is critical for preserving intestinal homeostasis. The barrier is formed by tightly linked intestinal epithelial cells (IECs) (i.e. enterocytes, goblet cells, neuroendocrine cells, tuft cells, Paneth cells, and M cells), which constantly self-renew and shed. IECs also communicate with microbiota, coordinate innate and adaptive effector cell functions. In this review, we summarize the signaling pathways contributing to intestinal cell fates and homeostasis functions. We focus especially on intestinal stem cell proliferation, cell junction formation, remodelling, hypoxia, the impact of intestinal microbiota, the immune system, inflammation, and metabolism. Recognizing the critical role of KRAS mutants in colorectal cancer, we highlight the connections of KRAS signaling pathways in coordinating these functions. Furthermore, we review the impact of KRAS colorectal cancer mutants on pathway rewiring associated with disruption and dysfunction of the normal intestinal homeostasis. Given that KRAS is still considered undruggable and the development of treatments that directly target KRAS are unlikely, we discuss the suitability of targeting pathways downstream of KRAS as well as alterations of cell extrinsic/microenvironmental factors as possible targets for modulating signaling pathways in colorectal cancer. Video Abstract.

Keywords:  Colorectal cancer; Hypoxia; Inflammation; Intestinal stem cells; KRAS; MAPK pathway; Metabolic reprogramming; Microenvironment; Reactive oxygen species; Small GTPases

DOI:  https://doi.org/10.1186/s12964-021-00712-3
Cancer Sci. 2021 Mar 08.

Therapeutic and prognostic implications of NOTCH and MAPK signaling in bladder cancer.

Gerald B Schulz, Sefer Elezkurtaj, Teresa Börding, Eva Marina Schmidt, Manal Elmasry, Christian G Stief, Thomas Kirchner, Alexander Karl, David Horst.

  Signaling pathways that drive bladder cancer (BC) progression may be promising and specific targets for systemic therapy. Here, we investigated the clinical significance and targetability of NOTCH and MAPK signaling for this aggressive malignancy. We assessed NOTCH1 and MAPK activity in 222 stage III and IV BC specimens of patients that had undergone radical cystectomy, and tested for clinical associations including cancer specific and overall survival. We examined therapeutic effects of NOTCH and MAPK repression in a murine xenograft model of human bladder cancer cells and evaluated tumor growth and tumor cell plasticity. In BC, NOTCH1 and MAPK signaling marked two distinct tumor cell subpopulations. The combination of high NOTCH1 and high MAPK activity indicated poor cancer specific and overall survival in univariate and multivariate analyses. Inhibition of NOTCH and MAPK in BC xenografts in vivo depleted targeted tumor cell subpopulations and revealed strong plasticity in signaling pathway activity. Combinatorial inhibition of NOTCH and MAPK signaling most strongly suppressed tumor growth. Our findings indicate that tumor cell subpopulations with high NOTCH and MAPK activity both contribute to tumor progression. Furthermore, we propose a new concept for BC therapy, which advocates specific and simultaneous targeting of these different tumor cell subpopulations through combined NOTCH and MAPK inhibition.

Keywords:  Biomarkers; Cystectomy; Mitogen-Activated Protein Kinase Kinases Receptors; Notch; Urinary Bladder Neoplasms

DOI:  https://doi.org/10.1111/cas.14878
Biochemistry (Mosc). 2020 Dec;85(12): 1570-1577

Nonphosphorylating Oxidation in Mitochondria and Related Processes.

D B Zorov, N V Andrianova, V A Babenko, L E Bakeeva, S D Zorov, L D Zorova, I B Pevsner, V A Popkov, E Yu Plotnikov, D N Silachev.

The mechanism of oxidative phosphorylation and its regulation remain one of the main problems of bioenergetics. Efficiency of the mitochondrial energization is determined by the relationship between the rate of generation of electrochemical potential of hydrogen ions and the rate of its expenditure on the synthesis of ATP and the use of ATP in endergonic reactions. Uncoupling (partial or complete), which occurs in the process of uncontrolled and controlled leakage of ions through the inner mitochondrial membrane, on the one hand leads to the decrease in the relative synthesis of ATP, and on the other, being consistent with the law of conservation of energy, leads to the formation of heat, generation of which is an essential function of the organism. In addition to increased thermogenesis, the increase of non-phosphorylating oxidation of various substrates is accompanied by the decrease in transmembrane potential, production of reactive oxygen species, and activation of oxygen consumption, water and carbon dioxide production, increase in the level of intracellular ADP and acidification of the cytosol. In this analysis, each of these factors will be considered separately for its role in regulating metabolism.

DOI: https://doi.org/10.1134/S0006297920120093
Lung Cancer. 2021 Mar 03. pii: S0169-5002(21)00095-7. [Epub ahead of print]154 151-160

Strategies to overcome resistance to immune checkpoint blockade in lung cancer.

Ilaria Attili, Paolo Tarantino, Antonio Passaro, Valeria Stati, Giuseppe Curigliano, Filippo de Marinis.

  The adoption of Immune checkpoint inhibitors (ICIs) allowed the achievement of impressive long-term survival results in non-small cell lung cancer (NSCLC), but most patients develop resistance to ICI treatment over time. Resistance to ICIs is mediated by several complex mechanisms affecting, but not limited to, tumour cell-intrinsic alterations and the tumour microenvironment. The possibility of modulating the immune response by interfering with specific alternative immune receptors, pathways and mediators might provide additional strategies to delay or prevent the development of resistance. Therefore, a greater in-depth investigation and understanding of these mechanisms aims to identify novel classes of immune targets and subsequently to evaluate potential new strategies for overcoming resistance, which will be assessed in this review.

Keywords:  Adoptive cell therapy; Gut microbiota; Immunotherapy; Resistance mechanisms; TIGIT; Tumour microenvironment

DOI:  https://doi.org/10.1016/j.lungcan.2021.02.035
Front Oncol. 2021 ;11 582694

Mitochondria's Role in the Maintenance of Cancer Stem Cells in Glioblastoma.

Yasaman Iranmanesh, Biao Jiang, Okoye C Favour, Zhangqi Dou, Jiawei Wu, Jinfan Li, Chongran Sun.

  Glioblastoma (GBM), one of the deadliest primary brain malignancies, is characterized by a high recurrence rate due to its limited response to existing therapeutic strategies such as chemotherapy, radiation therapy, and surgery. Several mechanisms and pathways have been identified to be responsible for GBM therapeutic resistance. Glioblastoma stem cells (GSCs) are known culprits of GBM resistance to therapy. GSCs are characterized by their unique self-renewal, differentiating capacity, and proliferative potential. They form a heterogeneous population of cancer stem cells within the tumor and are further divided into different subpopulations. Their distinct molecular, genetic, dynamic, and metabolic features distinguish them from neural stem cells (NSCs) and differentiated GBM cells. Novel therapeutic strategies targeting GSCs could effectively reduce the tumor-initiating potential, hence, a thorough understanding of mechanisms involved in maintaining GSCs' stemness cannot be overemphasized. The mitochondrion, a regulator of cellular physiological processes such as autophagy, cellular respiration, reactive oxygen species (ROS) generation, apoptosis, DNA repair, and cell cycle control, has been implicated in various malignancies (for instance, breast, lung, and prostate cancer). Besides, the role of mitochondria in GBM has been extensively studied. For example, when stressors, such as irradiation and hypoxia are present, GSCs utilize specific cytoprotective mechanisms like the activation of mitochondrial stress pathways to survive the harsh environment. Proliferating GBM cells exhibit increased cytoplasmic glycolysis in comparison to terminally differentiated GBM cells and quiescent GSCs that rely more on oxidative phosphorylation (OXPHOS). Furthermore, the Warburg effect, which is characterized by increased tumor cell glycolysis and decreased mitochondrial metabolism in the presence of oxygen, has been observed in GBM. Herein, we highlight the importance of mitochondria in the maintenance of GSCs.

Keywords:  GSC; glioblastoma; metabolism; mitochondria; quiescence; stem cell; stemness

DOI:  https://doi.org/10.3389/fonc.2021.582694
Cancer Res. 2021 Mar 09. pii: canres.1954.2020. [Epub ahead of print]

Fat induces glucose metabolism in non-transformed liver cells and promotes liver tumorigenesis.

Lindsay A Broadfield, João André Gonçalves Duarte, Roberta Schmieder, Dorien Broekaert, Koen Veys, Mélanie Planque, Kim Vriens, Yasuaki Karasawa, Francesco Napolitano, Suguru Fujita, Masashi Fujii, Miki Eto, Bryan Holvoet, Roman Vangoitsenhoven, Juan Fernandez-Garcia, Joke Van Elsen, Jonas Dehairs, Jia Zeng, James Dooley, Rebeca Alba Rubio, Jos van Pelt, Thomas G P Grünewald, Adrian Liston, Chantal Mathieu, Christophe M Deroose, Johannes V Swinnen, Diether Lambrechts, Diego di Bernardo, Shinya Kuroda, Katrien De Bock, Sarah-Maria Fendt.

Hepatic fat accumulation is associated with diabetes and hepatocellular carcinoma (HCC). Here we characterize the metabolic response that high fat availability elicits in livers prior to disease development. After a short term on a high fat diet, otherwise healthy mice showed elevated hepatic glucose uptake and increased glucose contribution to serine and pyruvate carboxylase activity compared to control diet mice. This glucose phenotype occurred independently from transcriptional or proteomic programming, which identifies increased peroxisomal and lipid metabolism pathways. High fat diet-fed mice exhibited increased lactate production when challenged with glucose. Consistently, administration of an oral glucose bolus to healthy individuals revealed a correlation between waist circumference and lactate secretion in a human cohort. In vitro, palmitate exposure stimulated production of reactive oxygen species and subsequent glucose uptake and lactate secretion in hepatocytes and liver cancer cells. Furthermore, high fat diet enhanced the formation of HCC compared to control diet in mice exposed to a hepatic carcinogen. Regardless of the dietary background, all murine tumors showed similar alterations in glucose metabolism to those identified in fat exposed non-transformed mouse livers; however, particular lipid species were elevated in high fat diet tumor and non-tumor-bearing high fat diet liver tissue. These findings suggest that fat can induce glucose-mediated metabolic changes in non-transformed liver cells similar to those found in HCC.

DOI: https://doi.org/10.1158/0008-5472.CAN-20-1954
Blood. 2021 Mar 08. pii: blood.2020006765. [Epub ahead of print]

Activation of the MAPK pathway mediates resistance to PI3K inhibitors in chronic lymphocytic leukemia (cll).

Ishwarya Murali, Siddha Kasar, Aishath Naeem, Svitlana Tyekucheva, Jasneet Kaur Khalsa, Emily M Thrash, Gilad Itchaki, Dimitri Livitz, Ignaty Leshchiner, Shuai Dong, Stacey Marie Fernandes, Gad Getz, Amy Johnson, Jennifer R Brown.

Inhibitors of Bruton's tyrosine kinase (BTKi) and phosphatidylinositol 3-kinase delta (PI3Kδi) that target the B cell receptor (BCR) signaling pathway have revolutionized the treatment of chronic lymphocytic leukemia (CLL). While mutations associated with resistance to BTK inhibitors have been identified, limited data are available on mechanisms of resistance to PI3Kδi. Here we present findings from longitudinal whole-exome sequencing of multiply relapsed CLL patients (Ncases=28) enrolled in PI3Ki trials. The non-responder subgroup was characterized by baseline activating mutations in MAP2K1, BRAF and KRAS in 60% of patients. PI3Kδ inhibition failed to inhibit ERK phosphorylation (pERK) in non-responder CLL cells with and without mutations, while treatment with MEKi rescued ERK inhibition. Overexpression of MAP2K1 mutants in vitro led to increased basal and inducible pERK and resistance to idelalisib. These data demonstrate that MAPK/ERK activation plays a key role in resistance to PI3Kδi in CLL and provide rationale for combination therapy with PI3Kδ and ERK inhibitors.

DOI: https://doi.org/10.1182/blood.2020006765
Nat Cancer. 2021 Feb;2(2): 141-156

Pathway-based classification of glioblastoma uncovers a mitochondrial subtype with therapeutic vulnerabilities.

Luciano Garofano, Simona Migliozzi, Young Taek Oh, Fulvio D'Angelo, Ryan D Najac, Aram Ko, Brulinda Frangaj, Francesca Pia Caruso, Kai Yu, Jinzhou Yuan, Wenting Zhao, Anna Luisa Di Stefano, Franck Bielle, Tao Jiang, Peter Sims, Mario L Suvà, Fuchou Tang, Xiao-Dong Su, Michele Ceccarelli, Marc Sanson, Anna Lasorella, Antonio Iavarone.

The transcriptomic classification of glioblastoma (GBM) has failed to predict survival and therapeutic vulnerabilities. A computational approach for unbiased identification of core biological traits of single cells and bulk tumors uncovered four tumor cell states and GBM subtypes distributed along neurodevelopmental and metabolic axes, classified as proliferative/progenitor, neuronal, mitochondrial and glycolytic/plurimetabolic. Each subtype was enriched with biologically coherent multiomic features. Mitochondrial GBM was associated with the most favorable clinical outcome. It relied exclusively on oxidative phosphorylation for energy production, whereas the glycolytic/plurimetabolic subtype was sustained by aerobic glycolysis and amino acid and lipid metabolism. Deletion of the glucose-proton symporter SLC45A1 was the truncal alteration most significantly associated with mitochondrial GBM, and the reintroduction of SLC45A1 in mitochondrial glioma cells induced acidification and loss of fitness. Mitochondrial, but not glycolytic/plurimetabolic, GBM exhibited marked vulnerability to inhibitors of oxidative phosphorylation. The pathway-based classification of GBM informs survival and enables precision targeting of cancer metabolism.

DOI: https://doi.org/10.1038/s43018-020-00159-4
Oncogene. 2021 Mar 12.

Proteasome 26S subunit, non-ATPases 1 (PSMD1) and 3 (PSMD3), play an oncogenic role in chronic myeloid leukemia by stabilizing nuclear factor-kappa B.

Alfonso E Bencomo-Alvarez, Andres J Rubio, Idaly M Olivas, Mayra A Gonzalez, Rebecca Ellwood, Carme Ripoll Fiol, Christopher A Eide, Joshua J Lara, Christian Barreto-Vargas, Luis F Jave-Suarez, Georgios Nteliopoulos, Alistair G Reid, Dragana Milojkovic, Brian J Druker, Jane Apperley, Jamshid S Khorashad, Anna M Eiring.

Tyrosine kinase inhibitors (TKIs) targeting BCR-ABL1 have revolutionized therapy for chronic myeloid leukemia (CML), paving the way for clinical development in other diseases. Despite success, targeting leukemic stem cells and overcoming drug resistance remain challenges for curative cancer therapy. To identify drivers of kinase-independent TKI resistance in CML, we performed genome-wide expression analyses on TKI-resistant versus sensitive CML cell lines, revealing a nuclear factor-kappa B (NF-κB) expression signature. Nucleocytoplasmic fractionation and luciferase reporter assays confirmed increased NF-κB activity in the nucleus of TKI-resistant versus sensitive CML cell lines and CD34+ patient samples. Two genes that were upregulated in TKI-resistant CML cells were proteasome 26S subunit, non-ATPases 1 (PSMD1) and 3 (PSMD3), both members of the 19S regulatory complex in the 26S proteasome. PSMD1 and PSMD3 were also identified as survival-critical genes in a published small hairpin RNA library screen of TKI resistance. We observed markedly higher levels of PSMD1 and PSMD3 mRNA in CML patients who had progressed to the blast phase compared with the chronic phase of the disease. Knockdown of PSMD1 or PSMD3 protein correlated with reduced survival and increased apoptosis in CML cells, but not in normal cord blood CD34+ progenitors. Luciferase reporter assays and immunoblot analyses demonstrated that PSMD1 and PSMD3 promote NF-κB protein expression in CML, and that signal transducer and activator of transcription 3 (STAT3) further activates NF-κB in scenarios of TKI resistance. Our data identify NF-κB as a transcriptional driver in TKI resistance, and implicate PSMD1 and PSMD3 as plausible therapeutic targets worthy of future investigation in CML and possibly other malignancies.

DOI: https://doi.org/10.1038/s41388-021-01732-6
Front Immunol. 2021 ;12 636238

PSGL-1 Immune Checkpoint Inhibition for CD4+ T Cell Cancer Immunotherapy.

Julia M DeRogatis, Karla M Viramontes, Emily N Neubert, Roberto Tinoco.

  Immune checkpoint inhibition targeting T cells has shown tremendous promise in the treatment of many cancer types and are now standard therapies for patients. While standard therapies have focused on PD-1 and CTLA-4 blockade, additional immune checkpoints have shown promise in promoting anti-tumor immunity. PSGL-1, primarily known for its role in cellular migration, has also been shown to function as a negative regulator of CD4+ T cells in numerous disease settings including cancer. PSGL-1 is highly expressed on T cells and can engage numerous ligands that impact signaling pathways, which may modulate CD4+ T cell differentiation and function. PSGL-1 engagement in the tumor microenvironment may promote CD4+ T cell exhaustion pathways that favor tumor growth. Here we highlight that blocking the PSGL-1 pathway on CD4+ T cells may represent a new cancer therapy approach to eradicate tumors.

Keywords:  CD4+ T cells; PSGL-1; anti-tumor immunity; cancer immunology; immune checkpoints

DOI:  https://doi.org/10.3389/fimmu.2021.636238
Cancer Res. 2021 Mar 08. pii: canres.1628.2020. [Epub ahead of print]

Targeting mitochondrial iron metabolism suppresses tumor growth and metastasis by inducing mitochondrial dysfunction and mitophagy.

Cristian Sandoval-Acuña, Natalia Torrealba, Veronika Tomkova, Sukanya Balkrishna Jadhav, Kristyna Blazkova, Ladislav Merta, Sandra Lettlova, Miroslava Kari Adamcová, Daniel Rosel, Jan Brábek, Jiri Neuzil, Jan Stursa, Lukas Werner, Jaroslav Truksa.

Deferoxamine (DFO) represents a widely used iron chelator for the treatment of iron overload. Here we describe the use of mitochondrially targeted deferoxamine (mitoDFO) as a novel approach to preferentially target cancer cells. The agent showed marked cytostatic, cytotoxic, and migrastatic properties in vitro, and it significantly suppressed tumor growth and metastasis in vivo. The underlying molecular mechanisms included (I) impairment of [Fe-S] cluster/heme biogenesis, leading to destabilization and loss of activity of [Fe-S] cluster/heme containing enzymes, (II) inhibition of mitochondrial respiration leading to mitochondrial ROS production, resulting in dysfunctional mitochondria with markedly reduced supercomplexes, and (III) fragmentation of the mitochondrial network and induction of mitophagy. Mitochondrial targeting of DFO represents a way to deprive cancer cells of biologically active iron, which is incompatible with their proliferation and invasion, without disrupting systemic iron metabolism. Our findings highlight the importance of mitochondrial iron metabolism for cancer cells and demonstrate repurposing deferoxamine into an effective anti-cancer drug via mitochondrial targeting.

DOI: https://doi.org/10.1158/0008-5472.CAN-20-1628
Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2021 Feb 28. pii: 1672-7347(2021)02-0113-08. [Epub ahead of print]46(2): 113-120

Oligomycin A promotes radioresistance in HT29 colorectal cancer cells and its mechanisms.

Xiaofei Li, Ruifang Tian, Lihui Wang, Cong Xu, Hui Wu, Lan Liu, Chenghui Huang.

   OBJECTIVES: Radiotherapy is one of the main therapies for colorectal cancer, but radioresistance often leads to radiotherapy failure. To improve the radioresistance, we explore the effect of oligomycin A, the H+-ATP synthase inhibitor, on the sensitivity of HT29 colorectal cancer cells to irradiation and its underlying mechanisms.
METHODS: The effects of different concentrations of oligomycin A on the survival rate and glycolysis of HT29 colorectal cancer cells at different time points were investigated via MTT and glycolysis assay. siRNA-PFK1 was synthesized in vitro and transfected into HT29 cells. The effects of oligomycin A on radiosensitivity of HT29 colorectal cancer cells were measured via MTT and colony formation assay. Western blotting was used to detect the effect of oligomycin A on the expression of glycolytic enzyme PFK1. We compared difference between the effects of siRNA-PFK1 group and oligomycin A combined with siRNA-PFK1 group on cell survival and glycolysis. After 4 Gy X-ray irradiation, the effects of cell survival and glycolysis between the siRNA-PFK1 group and the oligomycin A combined with siRNA-PFK1 group were compared.
RESULTS: Compared with the 0 μmol/L oligomycin A group, the cell survival rate of HT29 cells treated with 4 μmol/L oligomycin A was significantly increased (P<0.05), and the glucose uptake, the lactic acid, and the ATP production were also significantly increased (all P<0.01). After X-ray irradiation at different doses (0, 2, 4, 6, and 8 Gy), the colony formation rate and cell survival rate of the 4 μmol/L oligomycin A treated group were significantly higher than those in the 0 μmol/L oligomycin A group (both P<0.01). The sensitization enhancement ratio of oligomycin A on HT29 colorectal cancer cells was 0.4886. The expression of PFK1 in the 4 μmol/L oligomycin A group was significantly higher than that in the 0 μmol/L oligomycin A group (P<0.001). The glycolysis level, colony formation rate, and cell survival rate of the siRNA-PFK1 HT29 cells group were significantly lower than those in the 0 μmol/L oligomycin A group (all P<0.05), while the results in the 4 μmol/L oligomycin A combined with siRNA-PFK1 group were significantly higher than those in the siRNA-PFK1 group (all P<0.001). After 4 Gy X-ray irradiation, the colony formation rate and cell survival rate in the siRNA-PFK1 group were decreased compared with those in the irradiation group (P<0.01 or P<0.001), while the results of the 4 μmol/L oligomycin A combined with siRNA-PFK1 group were significantly higher than those in the siRNA-PFK1 group (both P<0.001).
CONCLUSIONS: Oligomycin A can promote the radioresistance of HT29 colorectal cancer cells, which may be related to up-regulation of the PFK1 expression and increase of cell glycolysis.

Keywords:  colorectal cancer; glycolysis; oligomycin A; phosphofructokinase 1; radiosensitivity

DOI:  https://doi.org/10.11817/j.issn.1672-7347.2021.200063
Sci Adv. 2021 Mar;pii: eabd6280. [Epub ahead of print]7(11):

Oxidative phosphorylation enhances the leukemogenic capacity and resistance to chemotherapy of B cell acute lymphoblastic leukemia.

Chiqi Chen, Xiaoxin Hao, Xiaoyun Lai, Ligen Liu, Jun Zhu, Hongfang Shao, Dan Huang, Hao Gu, Tinghua Zhang, Zhuo Yu, Li Xie, Xiaocui Zhang, Yi Yang, Jun Xu, Yuzheng Zhao, Zhigang Lu, Junke Zheng.

How metabolic status controls the fates of different types of leukemia cells remains elusive. Using a SoNar-transgenic mouse line, we demonstrated that B cell acute lymphoblastic leukemia (B-ALL) cells had a preference in using oxidative phosphorylation. B-ALL cells with a low SoNar ratio (SoNar-low) had enhanced mitochondrial respiration capacity, mainly resided in the vascular niche, and were enriched with more functional leukemia-initiating cells than that of SoNar-high cells in a murine B-ALL model. The SoNar-low cells were more resistant to cytosine arabinoside (Ara-C) treatment. cyclic adenosine 3',5'-monophosphate response element-binding protein transactivated pyruvate dehydrogenase complex component X and cytidine deaminase to maintain the oxidative phosphorylation level and Ara-C-induced resistance. SoNar-low human primary B-ALL cells also had a preference for oxidative phosphorylation. Suppressing oxidative phosphorylation with several drugs sufficiently attenuated Ara-C-induced resistance. Our study provides a unique angle for understanding the potential connections between metabolism and B-ALL cell fates.

DOI: https://doi.org/10.1126/sciadv.abd6280
Blood Cancer Discov. 2021 Mar;2(2): 146-161

A Therapeutic Strategy for Preferential Targeting of TET2 Mutant and TET-dioxygenase Deficient Cells in Myeloid Neoplasms.

Yihong Guan, Anand D Tiwari, James G Phillips, Metis Hasipek, Dale R Grabowski, Simona Pagliuca, Priyanka Gopal, Cassandra M Kerr, Vera Adema, Tomas Radivoyevitch, Yvonne Parker, Daniel J Lindner, Manja Meggendorfer, Mohamed Abazeed, Mikkeal A Sekeres, Omar Y Mian, Torsten Haferlach, Jaroslaw P Maciejewski, Babal K Jha.

  TET2 is frequently mutated in myeloid neoplasms. Genetic TET2 deficiency leads to skewed myeloid differentiation and clonal expansion, but minimal residual TET activity is critical for survival of neoplastic progenitor and stem cells. Consistent with mutual exclusivity of TET2 and neomorphic IDH1/2 mutations, here we report that IDH1/2 mutant-derived 2-hydroxyglutarate is synthetically lethal to TET-dioxygenase deficient cells. In addition, a TET-selective small molecule inhibitor decreased cytosine hydroxymethylation and restricted clonal outgrowth of TET2 mutant, but not normal hematopoietic precursor cells in vitro and in vivo. While TET-inhibitor phenocopied somatic TET2 mutations, its pharmacologic effects on normal stem cells were, unlike mutations, reversible. Treatment with TET inhibitor suppressed the clonal evolution of TET2 mutant cells in murine models and TET2-mutated human leukemia xenografts. These results suggest that TET inhibitors may constitute a new class of targeted agents in TET2 mutant neoplasia.

Keywords:  2-hydroxygluterate; 5hmC; IDH; MDS; TET2; α-ketoglutarate

DOI:  https://doi.org/10.1158/2643-3230.BCD-20-0173
Trends Cell Biol. 2021 Mar 04. pii: S0962-8924(21)00028-3. [Epub ahead of print]

Balancing ER-Mitochondrial Ca2+ Fluxes in Health and Disease.

Jens Loncke, Allen Kaasik, Ilya Bezprozvanny, Jan B Parys, Martijn Kerkhofs, Geert Bultynck.

  Organelles cooperate with each other to control cellular homeostasis and cell functions by forming close connections through membrane contact sites. Important contacts are present between the endoplasmic reticulum (ER), the main intracellular Ca2+-storage organelle, and the mitochondria, the organelle responsible not only for the majority of cellular ATP production but also for switching on cell death processes. Several Ca2+-transport systems focalize at these contact sites, thereby enabling the efficient transmission of Ca2+ signals from the ER toward mitochondria. This provides tight control of mitochondrial functions at the microdomain level. Here, we discuss how ER-mitochondrial Ca2+ transfers support cell function and how their dysregulation underlies, drives, or contributes to pathogenesis and pathophysiology, with a major focus on cancer and neurodegeneration but also with attention to other diseases such as diabetes and rare genetic diseases.

Keywords:  Ca(2+) signaling; MAMs; cancer; contact sites; genetic diseases; neurodegeneration

DOI:  https://doi.org/10.1016/j.tcb.2021.02.003