bims-mibica Biomed News
on Mitochondrial bioenergetics in cancer
Issue of 2021–08–15
fifty papers selected by
Kelsey Fisher-Wellman, East Carolina University
  1. GOT1 inhibition promotes pancreatic cancer cell death by ferroptosis.
  2. Effect of Novel Antipsychotics on Energy Metabolism - In Vitro Study in Pig Brain Mitochondria.
  3. Defining the molecular mechanisms of the mitochondrial permeability transition through genetic manipulation of F-ATP synthase.
  4. Energy Metabolic Plasticity of Colorectal Cancer Cells as a Determinant of Tumor Growth and Metastasis.
  5. PERM1 interacts with the MICOS-MIB complex to connect the mitochondria and sarcolemma via ankyrin B.
  6. ATPAF1 deficiency impairs ATP synthase assembly and mitochondrial respiration.
  7. Acetyl-CoA-driven respiration in frozen muscle contributes to the diagnosis of mitochondrial disease.
  8. The requirement for pyruvate dehydrogenase in leukemogenesis depends on cell lineage.
  9. Metabolic impairment of non-small cell lung cancers by mitochondrial HSPD1 targeting.
  10. Reduced Shmt2 Expression Impairs Mitochondrial Folate Accumulation and Respiration, and Leads to Uracil Accumulation in Mouse Mitochondrial DNA.
  11. Enhanced Sensitivity of Nonsmall Cell Lung Cancer with Acquired Resistance to Epidermal Growth Factor Receptor-Tyrosine Kinase Inhibitors to Phenformin: The Roles of a Metabolic Shift to Oxidative Phosphorylation and Redox Balance.
  12. Sequential Enzyme-Activated Macrotheranostic Probe for Selective Tumor Mitochondria Targeting.
  13. Allostatic hypermetabolic response in PGC1α/β heterozygote mouse despite mitochondrial defects.
  14. Elesclomol induces copper-dependent ferroptosis in colorectal cancer cells via degradation of ATP7A.
  15. 3D neuronal mitochondrial morphology in axons, dendrites, and somata of the aging mouse hippocampus.
  16. Integration of chronological omics data reveals mitochondrial regulatory mechanisms during the development of hepatocellular carcinoma.
  17. Advances in Mitochondrial Medicine and Translational Research.
  18. Suppressed mitochondrial respiration via NOX5-mediated redox imbalance contributes to the antitumor activity of anlotinib in oral squamous cell carcinoma.
  19. Aberrant NAD+ metabolism underlies Zika virus-induced microcephaly.
  20. Targeting Pin1 renders pancreatic cancer eradicable by synergizing with immunochemotherapy.
  21. Myoglobin, expressed in brown adipose tissue of mice, regulates the content and activity of mitochondria and lipid droplets.
  22. Pre-clinical evidence that salinomycin is active against retinoblastoma via inducing mitochondrial dysfunction, oxidative damage and AMPK activation.
  23. Potent Synergistic Effect on c-Myc Driven Colorectal Cancers Using a Novel Indole-Substituted Quinoline with a Plk1 Inhibitor.
  24. Disturbed mitochondrial acetylation in accordance with the availability of acetyl groups in hepatocellular carcinoma.
  25. Genome-scale integration of transcriptome and metabolome unveils squalene synthase and dihydrofolate reductase as targets against AML cells resistant to chemotherapy.
  26. Erythroid mitochondrial retention triggers myeloid-dependent type I interferon in human SLE.
  27. Nicotinamide riboside has minimal impact on energy metabolism in mouse models of mild obesity.
  28. The tumor suppressor folliculin inhibits lactate dehydrogenase A and regulates the Warburg effect.
  29. NAD+ supplement potentiates tumor-killing function by rescuing defective TUB-mediated NAMPT transcription in tumor-infiltrated T cells.
  30. The m6A methyltransferase METTL3 modifies PGC-1α mRNA promoting mitochondrial dysfunction and oxLDL-induced inflammation in monocytes.
  31. Two parallel pathways connect glutamine metabolism and mTORC1 activity to regulate glutamoptosis.
  32. Dissecting the concordant and disparate roles of NDUFAF3 and NDUFAF4 in mitochondrial complex I biogenesis.
  33. Cycling cancer persister cells arise from lineages with distinct programs.
  34. ME2 Promotes Proneural-Mesenchymal Transition and Lipogenesis in Glioblastoma.
  35. Targeting of eIF6-driven translation induces a metabolic rewiring that reduces NAFLD and the consequent evolution to hepatocellular carcinoma.
  36. Pharmacologically targeting molecular motor promotes mitochondrial fission for anti-cancer.
  37. Heterogeneous response of cancer-associated fibroblasts to the glucose deprivation through mitochondrial calcium uniporter.
  38. An SLC1A5 Variant Transports Glutamine to Mitochondria in Cancer Cells.
  39. Colorectal Cancer Cell Differentiation Is Dependent on the Repression of Aerobic Glycolysis by NDRG2-TXNIP Axis.
  40. Metabolism and Innate Immunity Meet at the Mitochondria.
  41. Stem cell factor SOX2 confers ferroptosis resistance in lung cancer via upregulation of SLC7A11.
  42. Mitochondrial Metabolism Targeted Nanoplatform for Efficient Triple-Negative Breast Cancer Combination Therapy.
  43. Spontaneous hydrolysis and spurious metabolic properties of α-ketoglutarate esters.
  44. ATXN7L3B promotes hepatocellular carcinoma stemness and is downregulated by metformin.
  45. Mito-Mendelian Interactions Alter in vivo Glucose Metabolism and Insulin Sensitivity in Healthy Mice.
  46. Intercellular Transfer of Mitochondria between Senescent Cells through Cytoskeleton-Supported Intercellular Bridges Requires mTOR and CDC42 Signalling.
  47. A proteomic and phosphoproteomic landscape of KRAS mutant cancers identifies combination therapies.
  48. IceR improves proteome coverage and data completeness in global and single-cell proteomics.
  49. The survival and proliferation of osteosarcoma cells are dependent on the mitochondrial BIG3-PHB2 complex formation.
  50. Interacting evolutionary pressures drive mutation dynamics and health outcomes in aging blood.
  1. Nat Commun. 2021 08 11. 12(1): 4860
    GOT1 inhibition promotes pancreatic cancer cell death by ferroptosis.
    Daniel M Kremer, Barbara S Nelson, Lin Lin, Emily L Yarosz, Christopher J Halbrook, Samuel A Kerk, Peter Sajjakulnukit, Amy Myers, Galloway Thurston, Sean W Hou, Eileen S Carpenter, Anthony C Andren, Zeribe C Nwosu, Nicholas Cusmano, Stephanie Wisner, Nneka E Mbah, Mengrou Shan, Nupur K Das, Brian Magnuson, Andrew C Little, Milan R Savani, Johanna Ramos, Tina Gao, Stephen A Sastra, Carmine F Palermo, Michael A Badgley, Li Zhang, John M Asara, Samuel K McBrayer, Marina Pasca di Magliano, Howard C Crawford, Yatrik M Shah, Kenneth P Olive, Costas A Lyssiotis.
      Cancer metabolism is rewired to support cell survival in response to intrinsic and environmental stressors. Identification of strategies to target these adaptions is an area of active research. We previously described a cytosolic aspartate aminotransaminase (GOT1)-driven pathway in pancreatic cancer used to maintain redox balance. Here, we sought to identify metabolic dependencies following GOT1 inhibition to exploit this feature of pancreatic cancer and to provide additional insight into regulation of redox metabolism. Using pharmacological methods, we identify cysteine, glutathione, and lipid antioxidant function as metabolic vulnerabilities following GOT1 withdrawal. We demonstrate that targeting any of these pathways triggers ferroptosis, an oxidative, iron-dependent form of cell death, in GOT1 knockdown cells. Mechanistically, we reveal that GOT1 inhibition represses mitochondrial metabolism and promotes a catabolic state. Consequently, we find that this enhances labile iron availability through autophagy, which potentiates the activity of ferroptotic stimuli. Overall, our study identifies a biochemical connection between GOT1, iron regulation, and ferroptosis.
    DOI:  https://doi.org/10.1038/s41467-021-24859-2
  2. Mol Neurobiol. 2021 Aug 08.
    Effect of Novel Antipsychotics on Energy Metabolism - In Vitro Study in Pig Brain Mitochondria.
    Matej Ľupták, Zdeněk Fišar, Jana Hroudová.
      The identification and quantification of mitochondrial effects of novel antipsychotics (brexpiprazole, cariprazine, loxapine, and lurasidone) were studied in vitro in pig brain mitochondria. Selected parameters of mitochondrial metabolism, electron transport chain (ETC) complexes, citrate synthase (CS), malate dehydrogenase (MDH), monoamine oxidase (MAO), mitochondrial respiration, and total ATP and reactive oxygen species (ROS) production were evaluated and associated with possible adverse effects of drugs. All tested antipsychotics decreased the ETC activities (except for complex IV, which increased in activity after brexpiprazole and loxapine addition). Both complex I- and complex II-linked respiration were dose-dependently inhibited, and significant correlations were found between complex I-linked respiration and both complex I activity (positive correlation) and complex IV activity (negative correlation). All drugs significantly decreased mitochondrial ATP production at higher concentrations. Hydrogen peroxide production was significantly increased at 10 µM brexpiprazole and lurasidone and at 100 µM cariprazine and loxapine. All antipsychotics acted as partial inhibitors of MAO-A, brexpiprazole and loxapine partially inhibited MAO-B. Based on our results, novel antipsychotics probably lacked oxygen uncoupling properties. The mitochondrial effects of novel antipsychotics might contribute on their adverse effects, which are mostly related to decreased ATP production and increased ROS production, while MAO-A inhibition might contribute to their antidepressant effect, and brexpiprazole- and loxapine-induced MAO-B inhibition might likely promote neuroplasticity and neuroprotection. The assessment of drug-induced mitochondrial dysfunctions is important in development of new drugs as well as in the understanding of molecular mechanism of adverse or side drug effects.
    Keywords:  ATP; Dopamine system stabilizers; Mitochondrial respiration; Monoamine oxidase; Oxidative phosphorylation; Reactive oxygen species
    DOI:  https://doi.org/10.1007/s12035-021-02498-4
  3. Nat Commun. 2021 08 10. 12(1): 4835
    Defining the molecular mechanisms of the mitochondrial permeability transition through genetic manipulation of F-ATP synthase.
    Andrea Carrer, Ludovica Tommasin, Justina Šileikytė, Francesco Ciscato, Riccardo Filadi, Andrea Urbani, Michael Forte, Andrea Rasola, Ildikò Szabò, Michela Carraro, Paolo Bernardi.
      F-ATP synthase is a leading candidate as the mitochondrial permeability transition pore (PTP) but the mechanism(s) leading to channel formation remain undefined. Here, to shed light on the structural requirements for PTP formation, we test cells ablated for g, OSCP and b subunits, and ρ0 cells lacking subunits a and A6L. Δg cells (that also lack subunit e) do not show PTP channel opening in intact cells or patch-clamped mitoplasts unless atractylate is added. Δb and ΔOSCP cells display currents insensitive to cyclosporin A but inhibited by bongkrekate, suggesting that the adenine nucleotide translocator (ANT) can contribute to channel formation in the absence of an assembled F-ATP synthase. Mitoplasts from ρ0 mitochondria display PTP currents indistinguishable from their wild-type counterparts. In this work, we show that peripheral stalk subunits are essential to turn the F-ATP synthase into the PTP and that the ANT provides mitochondria with a distinct permeability pathway.
    DOI:  https://doi.org/10.1038/s41467-021-25161-x
  4. Front Oncol. 2021 ;11 698951
    Energy Metabolic Plasticity of Colorectal Cancer Cells as a Determinant of Tumor Growth and Metastasis.
    Leenu Reinsalu, Marju Puurand, Vladimir Chekulayev, Sten Miller, Igor Shevchuk, Kersti Tepp, Egle Rebane-Klemm, Natalja Timohhina, Anton Terasmaa, Tuuli Kaambre.
      Metabolic plasticity is the ability of the cell to adjust its metabolism to changes in environmental conditions. Increased metabolic plasticity is a defining characteristic of cancer cells, which gives them the advantage of survival and a higher proliferative capacity. Here we review some functional features of metabolic plasticity of colorectal cancer cells (CRC). Metabolic plasticity is characterized by changes in adenine nucleotide transport across the outer mitochondrial membrane. Voltage-dependent anion channel (VDAC) is the main protein involved in the transport of adenine nucleotides, and its regulation is impaired in CRC cells. Apparent affinity for ADP is a functional parameter that characterizes VDAC permeability and provides an integrated assessment of cell metabolic state. VDAC permeability can be adjusted via its interactions with other proteins, such as hexokinase and tubulin. Also, the redox conditions inside a cancer cell may alter VDAC function, resulting in enhanced metabolic plasticity. In addition, a cancer cell shows reprogrammed energy transfer circuits such as adenylate kinase (AK) and creatine kinase (CK) pathway. Knowledge of the mechanism of metabolic plasticity will improve our understanding of colorectal carcinogenesis.
    Keywords:  VDAC; adenylate kinase; aerobic glycolysis; creatine kinase; mitochondria; oxidative phosphorylation; tumor energy metabolism
    DOI:  https://doi.org/10.3389/fonc.2021.698951
  5. Nat Commun. 2021 08 12. 12(1): 4900
    PERM1 interacts with the MICOS-MIB complex to connect the mitochondria and sarcolemma via ankyrin B.
    Theresa Bock, Clara Türk, Sriram Aravamudhan, Lena Keufgens, Wilhelm Bloch, Dieu Hien Rozsivalova, Vanina Romanello, Leonardo Nogara, Bert Blaauw, Aleksandra Trifunovic, Thomas Braun, Marcus Krüger.
      Skeletal muscle subsarcolemmal mitochondria (SSM) and intermyofibrillar mitochondria subpopulations have distinct metabolic activity and sensitivity, though the mechanisms that localize SSM to peripheral areas of muscle fibers are poorly understood. A protein interaction study and complexome profiling identifies PERM1 interacts with the MICOS-MIB complex. Ablation of Perm1 in mice reduces muscle force, decreases mitochondrial membrane potential and complex I activity, and reduces the numbers of SSM in skeletal muscle. We demonstrate PERM1 interacts with the intracellular adaptor protein ankyrin B (ANKB) that connects the cytoskeleton to the plasma membrane. Moreover, we identify a C-terminal transmembrane helix that anchors PERM1 into the outer mitochondrial membrane. We conclude PERM1 functions in the MICOS-MIB complex and acts as an adapter to connect the mitochondria with the sarcolemma via ANKB.
    DOI:  https://doi.org/10.1038/s41467-021-25185-3
  6. Mitochondrion. 2021 Aug 07. pii: S1567-7249(21)00106-9. [Epub ahead of print]
    ATPAF1 deficiency impairs ATP synthase assembly and mitochondrial respiration.
    Zhou Zhou, Kailiang Zhang, Zhiheng Liu, Xu Gao, Kai Huang, Chen Chen, Wang Daowen, Qinglin Yang, Qinqiang Long.
      ATP11p and ATP12p are two nuclear-encoded mitochondrial chaperone proteins required for assembling the F1Fo-ATP synthase F1 sector. ATPAF1 and ATPAF2 are the mammalian homologs of ATP11p and ATP12p. However, the biochemical and physiological relevance of ATPAF1 and ATPAF2 in animal tissues with high energy-dependence remains unclear. To explore the in vivo role of ATP assembly and the effects of ATP synthase deficiency in animals, we have generated knockout (KO) mouse models of these assembly factors using CRISPR/Cas9 technology. While the Atpaf2-KO mice were embryonically lethal, Atpaf1-KO mice grew to adulthood but with smaller body sizes and elevated blood lactate later in life. We specifically investigated how ATPAF1 deficiency may affect ATP synthase biogenesis and mitochondrial respiration in the mouse heart, an organ highly energy-dependent. Western blots and Blue-Native electrophoresis (BN-PAGE) demonstrated a decreased F1 content and ATP synthase dimers in the Atpaf1-KO heart. Mitochondria from ATPAF1-deficient hearts showed ultrastructural abnormalities with condensed degenerated mitochondria, loss of cristae, and impaired respiratory capacity. ATP synthase deficiency also leads to impaired autophagy and mitochondrial dynamic. Consequently, decreased cardiac function was exhibited in adult Atpaf1-KO mice. The results provide strong support that ATPAF1 is essential for ATP synthase assembly and mitochondrial oxidative phosphorylation, thus playing a crucial role in maintaining cardiac structure and function in animals.
    Keywords:  ATP synthase assembly; mitochondria; mitochondrial dysfunction; oxidative phosphorylation (OXPHOS)
    DOI:  https://doi.org/10.1016/j.mito.2021.08.005
  7. Eur J Clin Invest. 2021 Sep;51(9): e13574
    Acetyl-CoA-driven respiration in frozen muscle contributes to the diagnosis of mitochondrial disease.
    Felippe Henrique Zuccolotto-Dos-Reis, Silvia Helena Andrião Escarso, Jackeline Souza Araujo, Enilza Maria Espreafico, Luciane Carla Alberici, Claudia Ferreira da Rosa Sobreira.
       BACKGROUND: Freezing human biopsies is common in clinical practice for storage. However, this technique disrupts mitochondrial membranes, hampering further analyses of respiratory function. To contribute to laboratorial diagnosis of mitochondrial diseases, this study sought to develop a respirometry approach using O2k (Oroboros Ins.) to measure the whole electron transport chain (ETC) activity in homogenates of frozen skeletal muscle biopsies.
    PATIENTS AND METHODS: We enrolled 16 patients submitted to muscle biopsy in the process of routine diagnostic investigation: four with mitochondrial disease and severe mitochondrial dysfunction; seven with exercise intolerance and multiple deletions of mitochondrial DNA, presenting mild to moderate mitochondrial dysfunction; five without mitochondrial disease, as controls. Whole homogenates of muscle fragments were prepared using grinder-type equipment. O2 consumption rates were normalized using citrate synthase activity.
    RESULTS: Transmission electron microscopy confirmed mitochondrial membrane discontinuation, indicating increased permeability of mitochondrial membranes in homogenates from frozen biopsies. O2 consumption rates in the presence of acetyl-CoA lead to maximum respiratory rates sensitive to rotenone, malonate and antimycin. This protocol of acetyl-CoA-driven respiration (ACoAR), applied in whole homogenates of frozen muscle, was sensitive enough to identify ETC abnormality, even in patients with mild to moderate mitochondrial dysfunction. We demonstrated adequate repeatability of ACoAR and found significant correlation between O2 consumption rates and enzyme activity assays of individual ETC complexes.
    CONCLUSIONS: We present preliminary data on a simple, low cost and reliable procedure to measure respiratory function in whole homogenates of frozen skeletal muscle biopsies, contributing to diagnosis of mitochondrial diseases in humans.
    Keywords:  acetyl-CoA-driven respiration; electron transport chain; frozen skeletal muscle biopsy; high-resolution respirometry; mitochondrial diseases; oxygen consumption rate
    DOI:  https://doi.org/10.1111/eci.13574
  8. Cell Metab. 2021 Aug 03. pii: S1550-4131(21)00332-6. [Epub ahead of print]
    The requirement for pyruvate dehydrogenase in leukemogenesis depends on cell lineage.
    Sojeong Jun, Swetha Mahesula, Thomas P Mathews, Misty S Martin-Sandoval, Zhiyu Zhao, Elena Piskounova, Michalis Agathocleous.
      Cancer cells are metabolically similar to their corresponding normal tissues. Differences between cancers and normal tissues may reflect reprogramming during transformation or maintenance of the metabolism of the specific normal cell type that originated the cancer. Here, we compare glucose metabolism in hematopoiesis and leukemia. Thymus T cell progenitors were glucose avid and oxidized more glucose in the tricarboxylic acid cycle through pyruvate dehydrogenase (PDH) as compared with other hematopoietic cells. PDH deletion decreased double-positive T cell progenitor cells but had no effect on hematopoietic stem cells, myeloid progenitors, or other hematopoietic cells. PDH deletion blocked the development of Pten-deficient T cell leukemia, but not the development of a Pten-deficient myeloid neoplasm. Therefore, the requirement for PDH in leukemia reflected the metabolism of the normal cell of origin independently of the driver genetic lesion. PDH was required to prevent pyruvate accumulation and maintain glutathione levels and redox homeostasis.
    Keywords:  T cell leukemia; double-positive thymocytes; glycolysis; hematopoietic stem cells; metabolism; pyruvate dehydrogenase; thymus
    DOI:  https://doi.org/10.1016/j.cmet.2021.07.016
  9. J Exp Clin Cancer Res. 2021 Aug 07. 40(1): 248
    Metabolic impairment of non-small cell lung cancers by mitochondrial HSPD1 targeting.
    Beatrice Parma, Vignesh Ramesh, Paradesi Naidu Gollavilli, Aarif Siddiqui, Luisa Pinna, Annemarie Schwab, Sabine Marschall, Shuman Zhang, Christian Pilarsky, Francesca Napoli, Marco Volante, Sophia Urbanczyk, Dirk Mielenz, Henrik Daa Schrøder, Marc Stemmler, Heiko Wurdak, Paolo Ceppi.
       BACKGROUND: The identification of novel targets is of paramount importance to develop more effective drugs and improve the treatment of non-small cell lung cancer (NSCLC), the leading cause of cancer-related deaths worldwide. Since cells alter their metabolic rewiring during tumorigenesis and along cancer progression, targeting key metabolic players and metabolism-associated proteins represents a valuable approach with a high therapeutic potential. Metabolic fitness relies on the functionality of heat shock proteins (HSPs), molecular chaperones that facilitate the correct folding of metabolism enzymes and their assembly in macromolecular structures.
    METHODS: Gene fitness was determined by bioinformatics analysis from available datasets from genetic screenings. HSPD1 expression was evaluated by immunohistochemistry from formalin-fixed paraffin-embedded tissues from NSCLC patients. Real-time proliferation assays with and without cytotoxicity reagents, colony formation assays and cell cycle analyses were used to monitor growth and drug sensitivity of different NSCLC cells in vitro. In vivo growth was monitored with subcutaneous injections in immune-deficient mice. Cell metabolic activity was analyzed through extracellular metabolic flux analysis. Specific knockouts were introduced by CRISPR/Cas9.
    RESULTS: We show heat shock protein family D member 1 (HSPD1 or HSP60) as a survival gene ubiquitously expressed in NSCLC and associated with poor patients' prognosis. HSPD1 knockdown or its chemical disruption by the small molecule KHS101 induces a drastic breakdown of oxidative phosphorylation, and suppresses cell proliferation both in vitro and in vivo. By combining drug profiling with transcriptomics and through a whole-genome CRISPR/Cas9 screen, we demonstrate that HSPD1-targeted anti-cancer effects are dependent on oxidative phosphorylation and validated molecular determinants of KHS101 sensitivity, in particular, the creatine-transporter SLC6A8 and the subunit of the cytochrome c oxidase complex COX5B.
    CONCLUSIONS: These results highlight mitochondrial metabolism as an attractive target and HSPD1 as a potential theranostic marker for developing therapies to combat NSCLC.
    Keywords:  HSPD1; KHS101; Metabolism; Non-small cell lung cancer; Targeting
    DOI:  https://doi.org/10.1186/s13046-021-02049-8
  10. J Nutr. 2021 Aug 12. pii: nxab211. [Epub ahead of print]
    Reduced Shmt2 Expression Impairs Mitochondrial Folate Accumulation and Respiration, and Leads to Uracil Accumulation in Mouse Mitochondrial DNA.
    Joanna L Fiddler, Yuwen Xiu, Jamie E Blum, Simon G Lamarre, Whitney N Phinney, Sally P Stabler, Margaret E Brosnan, John T Brosnan, Anna E Thalacker-Mercer, Martha S Field.
       BACKGROUND: Adequate cellular thymidylate (dTMP) pools are essential for preservation of nuclear and mitochondrial genome stability. Previous studies have indicated that disruption in nuclear dTMP synthesis leads to increased uracil misincorporation into DNA, affecting genome stability. To date, the effects of impaired mitochondrial dTMP synthesis in nontransformed tissues have been understudied.
    OBJECTIVES: This study aimed to determine the effects of decreased serine hydroxymethyltransferase 2 (Shmt2) expression and dietary folate deficiency on mitochondrial DNA (mtDNA) integrity and mitochondrial function in mouse tissues.
    METHODS: Liver mtDNA content, and uracil content in liver mtDNA, were measured in Shmt2+/- and Shmt2+/+ mice weaned onto either a folate-sufficient control diet (2 mg/kg folic acid; C) or a modified diet lacking folic acid (0 mg/kg folic acid) for 7 wk. Shmt2+/- and Shmt2+/+ mouse embryonic fibroblast (MEF) cells were cultured in defined culture medium containing either 0 or 25 nM folate (6S-5-formyl-tetrahydrofolate, folinate) to assess proliferative capacity and mitochondrial function. Chi-square tests, linear mixed models, and 2-factor ANOVA with Tukey post hoc analyses were used to analyze data.
    RESULTS: Shmt2 +/- mice exhibited a 48%-67% reduction in SHMT2 protein concentrations in tissues. Interestingly, Shmt2+/- mice consuming the folate-sufficient C diet exhibited a 25% reduction in total folate in liver mitochondria. There was also a >20-fold increase in uracil in liver mtDNA in Shmt2+/- mice consuming the C diet, and dietary folate deficiency also increased uracil content in mouse liver mtDNA from both Shmt2+/+ and Shmt2+/- mice. Furthermore, decreased Shmt2 expression in MEF cells reduced cell proliferation, mitochondrial membrane potential, and oxygen consumption rate.
    CONCLUSIONS: This study demonstrates that Shmt2 heterozygosity and dietary folate deficiency impair mitochondrial dTMP synthesis in mice, as evidenced by the increased uracil in mtDNA. In addition, Shmt2 heterozygosity impairs mitochondrial function in MEF cells. These findings suggest that elevated uracil in mtDNA may impair mitochondrial function.
    Keywords:  SHMT2; folate; one-carbon metabolism; oxygen consumption rate; thymidylate; uracil
    DOI:  https://doi.org/10.1093/jn/nxab211
  11. Oxid Med Cell Longev. 2021 ;2021 5428364
    Enhanced Sensitivity of Nonsmall Cell Lung Cancer with Acquired Resistance to Epidermal Growth Factor Receptor-Tyrosine Kinase Inhibitors to Phenformin: The Roles of a Metabolic Shift to Oxidative Phosphorylation and Redox Balance.
    Suntae Kim, Ji Hye Im, Wan Kyu Kim, Young Jae Choi, Ji-Yoon Lee, Sang Kyum Kim, Sun Jo Kim, Sung Won Kwon, Keon Wook Kang.
       Background: Although the efficacy of epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR- TKI) therapy has been proven in non-small cell lung cancer (NSCLC) patients, acquired resistance to EGFR-TKIs presents a serious clinical problem. Hence, the identification of new therapeutic strategy is needed to treat EGFR-TKI-resistant NSCLC.
    Methods: Acquired EGFR-TKI-resistant lung cancer cell lines (HCC827, H1993, and H292 cells with acquired resistance to gefitinib or erlotinib) were used for cell-based studies. IncuCyte live cell analysis system and XFp analyzer were used for the determination of cell proliferation and energy metabolism, respectively. In vivo anticancer effect of phenformin was assessed in xenografts implanting HCC827 and gefitinib-resistant HCC827 (HCC827 GR) cells.
    Results: HCC827 GR and erlotinib-resistant H1993 (H1993 ER) cells exhibited different metabolic properties compared with their respective parental cells, HCC827, and H1993. In EGFR-TKI-resistant NSCLC cells, glycolysis markers including the glucose consumption rate, intracellular lactate level, and extracellular acidification rate were decreased; however, mitochondrial oxidative phosphorylation (OXPHOS) markers including mitochondria-driven ATP production, mitochondrial membrane potential, and maximal OXPHOS capacity were increased. Cell proliferation and tumor growth were strongly inhibited by biguanide phenformin via targeting of mitochondrial OXPHOS complex 1 in EGFR-TKI-resistant NSCLC cells. Inhibition of OXPHOS resulted in a reduced NAD+/NADH ratio and intracellular aspartate levels. Recovery of glycolysis by hexokinase 2 overexpression in erlotinib-resistant H292 (H292 ER) cells significantly reduced the anticancer effects of phenformin.
    Conclusion: Long-term treatment with EGFR-TKIs causes reactivation of mitochondrial metabolism, resulting in vulnerability to OXPHOS inhibitor such as phenformin. We propose a new therapeutic option for NSCLC with acquired EGFR-TKI resistance that focuses on cancer metabolism.
    DOI:  https://doi.org/10.1155/2021/5428364
  12. Acta Biomater. 2021 Aug 06. pii: S1742-7061(21)00515-8. [Epub ahead of print]
    Sequential Enzyme-Activated Macrotheranostic Probe for Selective Tumor Mitochondria Targeting.
    Di Ma, Qingyu Zong, Yuchen Du, Fangzhou Yu, Xuan Xiao, Rong Sun, Yuan Guo, Xinhua Wei, Youyong Yuan.
      Subcellular organelle targeted imaging and therapy are of enormous interest in cancer theranostics. However, the lack of tumor-selective organelle targeting has compromised their efficacy and safety. In this work, we found that the near-infrared (NIR) fluorophore hemicyanine (CyNH2) can selectively target mitochondria with strong cytotoxicity through decreasing the mitochondrial membrane potential and increasing the intracellular reactive oxygen species (ROS) levels. A macrotheranostic probe (denoted as PLCy) based on conjugating CyNH2 with an acetylated lysine group was developed with masked fluorescence and cytotoxicity, which could both be unmasked through sequential activation by cancer cells overexpressing histone deacetylases (HDACs) and cathepsin L (CTSL) enzymes for selective cancer cell mitochondria-targeted imaging and therapy. In vitro and in vivo studies confirmed that the specific fluorescence turn-on and toxicity were restored in cancer cells and efficiently inhibited tumor growth. This macrotheranostic probe with sequential enzyme activation and mitochondrial targeting is expected to have promising applications in imaging-guided cancer therapy with high specificity and efficiency.
    Keywords:  fluorescent probe; hemicyanine; mitochondria; theranostic
    DOI:  https://doi.org/10.1016/j.actbio.2021.08.002
  13. FASEB J. 2021 Sep;35(9): e21752
    Allostatic hypermetabolic response in PGC1α/β heterozygote mouse despite mitochondrial defects.
    Sergio Rodriguez-Cuenca, Christopher J Lelliot, Mark Campbell, Gopal Peddinti, Maite Martinez-Uña, Camilla Ingvorsen, Ana Rita Dias, Joana Relat, Silvia Mora, Tuulia Hyötyläinen, Antonio Zorzano, Matej Orešič, Mikael Bjursell, Mohammad Bohlooly-Y, Daniel Lindén, Antonio Vidal-Puig.
      Aging, obesity, and insulin resistance are associated with low levels of PGC1α and PGC1β coactivators and defective mitochondrial function. We studied mice deficient for PGC1α and PGC1β [double heterozygous (DH)] to investigate their combined pathogenic contribution. Contrary to our hypothesis, DH mice were leaner, had increased energy dissipation, a pro-thermogenic profile in BAT and WAT, and improved carbohydrate metabolism compared to wild types. WAT showed upregulation of mitochondriogenesis/oxphos machinery upon allelic compensation of PGC1α4 from the remaining allele. However, DH mice had decreased mitochondrial OXPHOS and biogenesis transcriptomes in mitochondria-rich organs. Despite being metabolically healthy, mitochondrial defects in DH mice impaired muscle fiber remodeling and caused qualitative changes in the hepatic lipidome. Our data evidence first the existence of organ-specific compensatory allostatic mechanisms are robust enough to drive an unexpected phenotype. Second, optimization of adipose tissue bioenergetics is sufficient to maintain a healthy metabolic phenotype despite a broad severe mitochondrial dysfunction in other relevant metabolic organs. Third, the decrease in PGC1s in adipose tissue of obese and diabetic patients is in contrast with the robustness of the compensatory upregulation in the adipose of the DH mice.
    Keywords:  PGC-1alpha; adipose tissue; hepatic lipidome; lipotoxicity; mitochondrial dysfunction
    DOI:  https://doi.org/10.1096/fj.202100262RR
  14. Mol Oncol. 2021 Aug 14.
    Elesclomol induces copper-dependent ferroptosis in colorectal cancer cells via degradation of ATP7A.
    Wei Gao, Zhao Huang, Jiufei Duan, Edouard C Nice, Jie Lin, Canhua Huang.
      Cancer cells reprogram their copper metabolism to adapt to adverse microenvironments, such as oxidative stress. The copper chelator elesclomol has been reported to have considerable anticancer efficacy, but the underlying mechanisms remain largely unknown. In this study, we found that elesclomol-mediated copper overload inhibits colorectal cancer both in vitro and in vivo. Elesclomol alone promotes the degradation of the copper transporter ATP7A, which retards the proliferation of colorectal cancer cells. This property distinguishes it from several other copper chelators. Combinational treatment of elesclomol and copper leads to copper retention within mitochondria due to ATP7A loss, leading to ROS accumulation, which in turn promotes the degradation of SLC7A11, thus further enhancing oxidative stress and consequent ferroptosis in CRC cells. This effect accounts for the robust antitumor activity of elesclomol against colorectal cancer, which can be reversed by the administration of antioxidants and ferroptosis inhibitors, as well as the overexpression of ATP7A. In summary, our findings indicate that elesclomol-induced copper chelation inhibits colorectal cancer by targeting ATP7A and regulating ferroptosis.
    Keywords:  ATP7A; Colorectal cancer; Copper; Elesclomol; Ferroptosis
    DOI:  https://doi.org/10.1002/1878-0261.13079
  15. Cell Rep. 2021 Aug 10. pii: S2211-1247(21)00939-6. [Epub ahead of print]36(6): 109509
    3D neuronal mitochondrial morphology in axons, dendrites, and somata of the aging mouse hippocampus.
    Julie Faitg, Clay Lacefield, Tracey Davey, Kathryn White, Ross Laws, Stylianos Kosmidis, Amy K Reeve, Eric R Kandel, Amy E Vincent, Martin Picard.
      The brain's ability to process complex information relies on the constant supply of energy through aerobic respiration by mitochondria. Neurons contain three anatomically distinct compartments-the soma, dendrites, and projecting axons-which have different energetic and biochemical requirements, as well as different mitochondrial morphologies in cultured systems. In this study, we apply quantitative three-dimensional electron microscopy to map mitochondrial network morphology and complexity in the mouse brain. We examine somatic, dendritic, and axonal mitochondria in the dentate gyrus and cornu ammonis 1 (CA1) of the mouse hippocampus, two subregions with distinct principal cell types and functions. We also establish compartment-specific differences in mitochondrial morphology across these cell types between young and old mice, highlighting differences in age-related morphological recalibrations. Overall, these data define the nature of the neuronal mitochondrial network in the mouse hippocampus, providing a foundation to examine the role of mitochondrial morpho-function in the aging brain.
    Keywords:  3D reconstruction; SBF-SEM; aging; hippocampus; microscopy; mitochondria; morphology; morphometry; three-dimensional; topology
    DOI:  https://doi.org/10.1016/j.celrep.2021.109509
  16. PLoS One. 2021 ;16(8): e0256016
    Integration of chronological omics data reveals mitochondrial regulatory mechanisms during the development of hepatocellular carcinoma.
    J Noé García-Chávez, Verónica R Vásquez-Garzón, Mercedes G López, Saúl Villa-Treviño, Rafael Montiel.
      Mitochondria participate in multiple functions in eukaryotic cells. Although disruption of mitochondrial function has been associated with energetic deregulation in cancer, the chronological changes in mitochondria during cancer development remain unclear. With the aim to assess the role of mitochondria throughout cancer development, we analyzed samples chronologically obtained from induced hepatocellular carcinoma (HCC) in rats. In our analyses, we integrated mitochondrial proteomic data, mitochondrial metabolomic data and nuclear genome transcriptomic data. We used pathway over-representation and weighted gene co-expression network analysis (WGCNA) to integrate expression profiles of genes, miRNAs, proteins and metabolite levels throughout HCC development. Our results show that mitochondria are dynamic organelles presenting specific modifications in different stages of HCC development. We also found that mitochondrial proteomic profiles from tissues adjacent to nodules or tumor are determined more by the stage of HCC development than by tissue type, and we evaluated two models to predict HCC stage of the samples using proteomic profiles. Finally, we propose an omics integration pipeline to massively identify molecular features that could be further evaluated as key regulators, biomarkers or therapeutic targets. As an example, we show a group of miRNAs and transcription factors as candidates, responsible for mitochondrial metabolic modification in HCC.
    DOI:  https://doi.org/10.1371/journal.pone.0256016
  17. Mitochondrion. 2021 Aug 04. pii: S1567-7249(21)00102-1. [Epub ahead of print]
    Advances in Mitochondrial Medicine and Translational Research.
    Raviprasad Kuthethur, Keshava Prasad, Sanjiban Chakrabarty, Shama Prasada Kabekkodu, Keshav K Singh, Kumarasamy Thangaraj, Kapaettu Satyamoorthy.
      Current knowledge of mitochondrial biology and function has provided with tools and technologies that helped a better understanding of the molecular etiology of complex mitochondrial disorders. Dual genetic control of this subcellular organelle function regulates various signaling mechanisms which are essential for metabolism, bioenergetics, fatty acid biosynthesis, and DNA replication & repair. Understanding nuclear mitochondrial crosstalk through advanced genomics as well as clinical perspectives is the overall basis of mitochondrial research and medicine, also the sole objective of Society for Mitochondrial Medicine and Research (SMRM) - India. The eighth virtual international conference on 'Advances in Mitochondrial Medicine and Translational Research' was organized at the Manipal School of Life Sciences, MAHE, Manipal, India, during 6 - 7 November 2020. The aim of the virtual conference was to highlight the recent advances and future perspectives that represent comprehensive clinical and fundamental research interests in the area of mitochondrial biology of human diseases. To systematically present the various findings in mitochondrial biology, the meeting was themed with specific aspects comprising (a) mitochondrial disorders: clinical & genomic perspectives, (b) mitochondria in cancer, (c) mitochondrial metabolism & disorders, and (d) mitochondrial diseases & therapy. This report provides an overview of the recent advancements in the area of mitochondrial biology and medicine that was discussed at the conference.
    Keywords:  Anterograde and retrograde signaling; Cancer and mitochondria; Mitochondrial disorders; Mitochondrial metabolism; Therapeutics and mitochondria; miRNAs and mitochondria
    DOI:  https://doi.org/10.1016/j.mito.2021.08.001
  18. J Genet Genomics. 2021 Jul 15. pii: S1673-8527(21)00201-0. [Epub ahead of print]
    Suppressed mitochondrial respiration via NOX5-mediated redox imbalance contributes to the antitumor activity of anlotinib in oral squamous cell carcinoma.
    Zhexun Huang, Qiao Su, Wuguo Li, Hui Ren, Huiqiang Huang, Anxun Wang.
      Anlotinib, a novel multitarget tyrosine kinase inhibitor, has shown promising results in the management of various carcinomas. This study aimed to investigate the antitumor activity of anlotinib in oral squamous cell carcinoma (OSCC) and the underlying molecular mechanism. A retrospective clinical study revealed that anlotinib improved the median progression-free survival (mPFS) and median overall survival (mOS) of patients with recurrent and metastatic (R/M) OSCC, respectively. Functional studies revealed that anlotinib markedly inhibited in vitro proliferation of OSCC cells and impeded in vivo tumor growth of OSCC patient-derived xenograft models. Mechanistically, RNA-sequencing identified that oxidative stress, oxidative phosphorylation and AKT/mTOR signaling were involved in anlotinib-treated OSCC cells. Anlotinib upregulated NADPH oxidase 5 (NOX5) expression, elevated reactive oxygen species (ROS) production, impaired mitochondrial respiration, and promoted apoptosis. Moreover, anlotinb also inhibited phospho-Akt (p-AKT) expression and elevated p-eIF2α expression in OSCC cells. NOX5 knockdown attenuated these inhibitory effects and cytotoxicity in anlotinib-treated OSCC cells. Collectively, we demonstrated that anlotinib monotherapy demonstrated favorable anticancer activity and manageable toxicities in patients with R/M OSCC. The antitumor activity of anlotinib in OSCC may be mainly involved in the suppression of mitochondrial respiration via NOX5-mediated redox imbalance and the AKT/eIF2α pathway.
    Keywords:  Anlotinib; Mitochondrial respiration function; NOX5; Oral squamous cell carcinoma; Oxidative phosphorylation; Oxidative stress
    DOI:  https://doi.org/10.1016/j.jgg.2021.06.014
  19. Nat Metab. 2021 Aug 12.
    Aberrant NAD+ metabolism underlies Zika virus-induced microcephaly.
    Huanhuan Pang, Yisheng Jiang, Jie Li, Yushen Wang, Meng Nie, Nan Xiao, Shuo Wang, Zhihong Song, Fansen Ji, Yafei Chang, Yu Zheng, Ke Yao, LiAng Yao, Shao Li, Peng Li, Lei Song, Xun Lan, Zhiheng Xu, Zeping Hu.
      Zika virus (ZIKV) infection during pregnancy can cause microcephaly in newborns, yet the underlying mechanisms remain largely unexplored. Here, we reveal extensive and large-scale metabolic reprogramming events in ZIKV-infected mouse brains by performing a multi-omics study comprising transcriptomics, proteomics, phosphoproteomics and metabolomics approaches. Our proteomics and metabolomics analyses uncover dramatic alteration of nicotinamide adenine dinucleotide (NAD+)-related metabolic pathways, including oxidative phosphorylation, TCA cycle and tryptophan metabolism. Phosphoproteomics analysis indicates that MAPK and cyclic GMP-protein kinase G signaling may be associated with ZIKV-induced microcephaly. Notably, we demonstrate the utility of our rich multi-omics datasets with follow-up in vivo experiments, which confirm that boosting NAD+ by NAD+ or nicotinamide riboside supplementation alleviates cell death and increases cortex thickness in ZIKV-infected mouse brains. Nicotinamide riboside supplementation increases the brain and body weight as well as improves the survival in ZIKV-infected mice. Our study provides a comprehensive resource of biological data to support future investigations of ZIKV-induced microcephaly and demonstrates that metabolic alterations can be potentially exploited for developing therapeutic strategies.
    DOI:  https://doi.org/10.1038/s42255-021-00437-0
  20. Cell. 2021 Aug 06. pii: S0092-8674(21)00879-5. [Epub ahead of print]
    Targeting Pin1 renders pancreatic cancer eradicable by synergizing with immunochemotherapy.
    Kazuhiro Koikawa, Shin Kibe, Futoshi Suizu, Nobufumi Sekino, Nami Kim, Theresa D Manz, Benika J Pinch, Dipikaa Akshinthala, Ana Verma, Giorgio Gaglia, Yutaka Nezu, Shizhong Ke, Chenxi Qiu, Kenoki Ohuchida, Yoshinao Oda, Tae Ho Lee, Babara Wegiel, John G Clohessy, Nir London, Sandro Santagata, Gerburg M Wulf, Manuel Hidalgo, Senthil K Muthuswamy, Masafumi Nakamura, Nathanael S Gray, Xiao Zhen Zhou, Kun Ping Lu.
      Pancreatic ductal adenocarcinoma (PDAC) is characterized by notorious resistance to current therapies attributed to inherent tumor heterogeneity and highly desmoplastic and immunosuppressive tumor microenvironment (TME). Unique proline isomerase Pin1 regulates multiple cancer pathways, but its role in the TME and cancer immunotherapy is unknown. Here, we find that Pin1 is overexpressed both in cancer cells and cancer-associated fibroblasts (CAFs) and correlates with poor survival in PDAC patients. Targeting Pin1 using clinically available drugs induces complete elimination or sustained remissions of aggressive PDAC by synergizing with anti-PD-1 and gemcitabine in diverse model systems. Mechanistically, Pin1 drives the desmoplastic and immunosuppressive TME by acting on CAFs and induces lysosomal degradation of the PD-1 ligand PD-L1 and the gemcitabine transporter ENT1 in cancer cells, besides activating multiple cancer pathways. Thus, Pin1 inhibition simultaneously blocks multiple cancer pathways, disrupts the desmoplastic and immunosuppressive TME, and upregulates PD-L1 and ENT1, rendering PDAC eradicable by immunochemotherapy.
    Keywords:  Pin1; cancer immune evasion; cancer-associated fibroblasts; chemotherapy; combination therapy; immuni checkpoint therapy; pancreatic cancer; targeted therapy; tumor immune microenvironment; tumor microenvironment
    DOI:  https://doi.org/10.1016/j.cell.2021.07.020
  21. Biochim Biophys Acta Mol Cell Biol Lipids. 2021 Aug 09. pii: S1388-1981(21)00154-2. [Epub ahead of print] 159026
    Myoglobin, expressed in brown adipose tissue of mice, regulates the content and activity of mitochondria and lipid droplets.
    Mostafa A Aboouf, Julia Armbruster, Markus Thiersch, Max Gassmann, Axel Gödecke, Erich Gnaiger, Glen Kristiansen, Anne Bicker, Thomas Hankeln, Hao Zhu, Thomas A Gorr.
      The identification of novel physiological regulators that stimulate energy expenditure through brown adipose tissue (BAT) activity in substrate catalysis is of utmost importance to understand and treat metabolic diseases. Myoglobin (MB), known to store or transport oxygen in heart and skeletal muscles, has recently been found to bind fatty acids with physiological constants in its oxygenated form (i.e., MBO2). Here, we investigated the in vivo effect of MB expression on BAT activity. In particular, we studied mitochondrial function and lipid metabolism as essential determinants of energy expenditure in this tissue. We show in a MB-null (MBko) mouse model that MB expression in BAT impacts on the activity of brown adipocytes in a twofold manner: i) by elevating mitochondrial density plus maximal respiration capacity, and through that, by stimulating BAT oxidative metabolism along with the organelles` uncoupled respiration; and ii) by influencing the free fatty acids pool towards a palmitate-enriched composition and shifting the lipid droplet (LD) equilibrium towards higher counts of smaller droplets. These metabolic changes were accompanied by the up-regulated expression of thermogenesis markers UCP1, CIDEA, CIDEC, PGC1-α and PPAR-α in the BAT of MB wildtype (MBwt) mice. Along with the emergence of the "browning" BAT morphology, MBwt mice exhibited a leaner phenotype when compared to MBko littermates at 20 weeks of age. Our data shed novel insights into MB's role in linking oxygen and lipid-based thermogenic metabolism. The findings suggest potential new strategies of targeting the MB pathway to treat metabolic disorders related to diminishing energy expenditure.
    Keywords:  Brown adipose tissue; Lipid droplet; Mitochondria; Myoglobin; OXPHOS; Respirometry
    DOI:  https://doi.org/10.1016/j.bbalip.2021.159026
  22. J Bioenerg Biomembr. 2021 Aug 08.
    Pre-clinical evidence that salinomycin is active against retinoblastoma via inducing mitochondrial dysfunction, oxidative damage and AMPK activation.
    Jing Li, Yao Min.
      The poor outcomes in retinoblastoma necessitate new treatments. Salinomycin is an attractive candidate, and has demonstrated selective anti-cancer properties in different cancer types. This work addressed the efficacy of salinomycin in retinoblastoma models and probe the associated mechanisms. Cellular functional assays were conducted to determine the effects salinomycin in vitro. Xenograft retinoblastoma mouse model was established to investigate the efficacy of salinomycin in vivo. Biochemical assays were conducted to analyze the mechanism of salinomycin's action focusing on mitochondrial functions, energy reduction-related signaling pathways. Salinomycin has positive effects towards retinoblastoma cells regardless of heterogeneity through suppressing growth and inducing apoptosis. Salinomycin also specifically inhibits cells displaying stemness and highly invasive phenotypes. Using retinoblastoma xenograft mouse model, we show that salinomycin at non-toxic dose effectively inhibits growth and induces apoptosis. Mechanistic studies show that salinomycin inhibits mitochondrial respiration via specifically suppressing complex I and II activities, reduces mitochondrial membrane potential and decreases energy reduction, followed by induction of oxidative stress and damage, AMPK activation and mTOR inhibition. Our study highlights that adding salinomycin to the existing treatment armamentarium for retinoblastoma is beneficial.
    Keywords:  AMPK/mTOR; Mitochondria respiration; Retinoblastoma; Salinomycin
    DOI:  https://doi.org/10.1007/s10863-021-09915-2
  23. Mol Cancer Ther. 2021 Aug 10. pii: molcanther.1017.2020. [Epub ahead of print]
    Potent Synergistic Effect on c-Myc Driven Colorectal Cancers Using a Novel Indole-Substituted Quinoline with a Plk1 Inhibitor.
    Yanqi Xie, Wen Zhang, Lichao Guo, Liliia M Kril, Kristin L Begley, Vitaliy M Sviripa, Xi Chen, Xifu Liu, Eun Y Lee, Daheng He, Chi Wang, Tianyan Gao, Xiaoqi Liu, B Mark Evers, David S Watt, Chunming Liu.
      Developing effective treatments for colorectal cancers through combinations of small-molecule approaches and immunotherapies present intriguing possibilities for managing these otherwise intractable cancers. During a broad-based, screening effort against multiple colorectal cancer cell lines, we identified indole-substituted quinolines (ISQs), such as N7,N7-dimethyl-3-(1-methyl-1H-indol-3-yl)quinoline-2,7-diamine (ISQ-1), as potent in vitro inhibitors of several cancer cell lines. We found that ISQ-1 inhibited Wnt signaling, a main driver in the pathway governing colorectal cancer development, and ISQ-1 also activated adenosine monophosphate kinase (AMPK), a cellular energy-homeostasis master regulator. We explored the effect of ISQs on cell metabolism. Seahorse assays measuring oxygen consumption rate (OCR) indicated that ISQ-1 inhibited complex I (i.e., NADH ubiquinone oxidoreductase) in the mitochondrial, electron transport chain (ETC). In addition, ISQ-1 treatment showed remarkable synergistic depletion of oncogenic c-Myc protein level in vitro and induced strong tumor remission in vivo when administered together with BI2536, a polo-like kinase-1 (Plk1) inhibitor. These studies point toward the potential value of dual drug therapies targeting the ETC and Plk-1 for the treatment of c-Myc-driven cancers.
    DOI:  https://doi.org/10.1158/1535-7163.MCT-20-1017
  24. Mitochondrion. 2021 Aug 07. pii: S1567-7249(21)00105-7. [Epub ahead of print]
    Disturbed mitochondrial acetylation in accordance with the availability of acetyl groups in hepatocellular carcinoma.
    Xiaowen Hu, Dandan Wang, Liya Sun, Yan Gao, Daizhan Zhou, Xuemei Tong, Jing Li, Hui Lin, Ying Qing, Shujiao Du, Xuhan Yang, Jie Jiang, Guoquan Yan, Zhiyun Wei, Qingyu Wang, Juan Zhang, Lin He, Chunling Wan.
      As an essential post-translational modification, acetylation participates in various cellular processes and shows aberrances during tumorigenesis. Owing to its modification substrate, acetyl-CoA, acetylation is postulated as a depot for acetyl groups and evolve to build a connection between epigenetics and metabolism. Here we depict a distinct acetylome atlas of hepatocellular carcinoma from the perspectives of both protein acetylation and acetyl-CoA metabolism. We found that tumor acetylome demonstrated a compartment-dependent alteration that the acetylation level of mitochondrial proteins tended to be decreased while nuclear proteins were highly acetylated. In addition, elevated expression of ATP-citrate synthase (ACLY) was observed in tumors, which would facilitate histone acetylation by transporting mitochondrial acetyl coenzyme A to the nucleus. A hypothetical model of the oncogenic acetylome was proposed that growing demands for histone acetylation in tumor cells would drive the relocalization of acetyl-CoA to the nucleus, which may contribute to the global deacetylation of mitochondrial proteins to support the nuclear acetyl-CoA pool in an ACLY-dependent manner. Our findings are thought-provoking on the potential linkage between epigenetics and metabolism in the progression of tumorigenesis.
    Keywords:  acetyl coenzyme A metabolism; compartment-characterized regulation; hepatocellular carcinoma; post-translational modification; protein acetylation; proteome
    DOI:  https://doi.org/10.1016/j.mito.2021.08.004
  25. Comput Struct Biotechnol J. 2021 ;19 4059-4066
    Genome-scale integration of transcriptome and metabolome unveils squalene synthase and dihydrofolate reductase as targets against AML cells resistant to chemotherapy.
    Effrosyni Karakitsou, Carles Foguet, Miriam G Contreras Mostazo, Nina Kurrle, Frank Schnütgen, Martin Michaelis, Jindrich Cinatl, Silvia Marin, Marta Cascante.
      The development of resistance to chemotherapeutic agents, such as Doxorubicin (DOX) and cytarabine (AraC), is one of the greatest challenges to the successful treatment of Acute Myeloid Leukemia (AML). Such acquisition is often underlined by a metabolic reprogramming that can provide a therapeutic opportunity, as it can lead to the emergence of vulnerabilities and dependencies to be exploited as targets against the resistant cells. In this regard, genome-scale metabolic models (GSMMs) have emerged as powerful tools to integrate multiple layers of data to build cancer-specific models and identify putative metabolic vulnerabilities. Here, we use genome-scale metabolic modelling to reconstruct a GSMM of the THP1 AML cell line and two derivative cell lines, one with acquired resistance to AraC and the second with acquired resistance to DOX. We also explore how, adding to the transcriptomic layer, the metabolomic layer enhances the selectivity of the resulting condition specific reconstructions. The resulting models enabled us to identify and experimentally validate that drug-resistant THP1 cells are sensitive to the FDA-approved antifolate methotrexate. Moreover, we discovered and validated that the resistant cell lines could be selectively targeted by inhibiting squalene synthase, providing a new and promising strategy to directly inhibit cholesterol synthesis in AML drug resistant cells.
    Keywords:  Acute Myeloid Leukemia (AML); Drug resistance; Drug targets; Metabolic models; Metabolic vulnerabilities
    DOI:  https://doi.org/10.1016/j.csbj.2021.06.049
  26. Cell. 2021 Aug 03. pii: S0092-8674(21)00880-1. [Epub ahead of print]
    Erythroid mitochondrial retention triggers myeloid-dependent type I interferon in human SLE.
    Simone Caielli, Jacob Cardenas, Adriana Almeida de Jesus, Jeanine Baisch, Lynnette Walters, Jean Philippe Blanck, Preetha Balasubramanian, Cristy Stagnar, Marina Ohouo, Seunghee Hong, Lorien Nassi, Katie Stewart, Julie Fuller, Jinghua Gu, Jacques F Banchereau, Tracey Wright, Raphaela Goldbach-Mansky, Virginia Pascual.
      Emerging evidence supports that mitochondrial dysfunction contributes to systemic lupus erythematosus (SLE) pathogenesis. Here we show that programmed mitochondrial removal, a hallmark of mammalian erythropoiesis, is defective in SLE. Specifically, we demonstrate that during human erythroid cell maturation, a hypoxia-inducible factor (HIF)-mediated metabolic switch is responsible for the activation of the ubiquitin-proteasome system (UPS), which precedes and is necessary for the autophagic removal of mitochondria. A defect in this pathway leads to accumulation of red blood cells (RBCs) carrying mitochondria (Mito+ RBCs) in SLE patients and in correlation with disease activity. Antibody-mediated internalization of Mito+ RBCs induces type I interferon (IFN) production through activation of cGAS in macrophages. Accordingly, SLE patients carrying both Mito+ RBCs and opsonizing antibodies display the highest levels of blood IFN-stimulated gene (ISG) signatures, a distinctive feature of SLE.
    Keywords:  CANDLE syndrome; HIF2a; autoimmunity; cGAS; human erythropoiesis; interferon; mitochondrial DNA; mitophagy; proteasome; systemic lupus erythematosus
    DOI:  https://doi.org/10.1016/j.cell.2021.07.021
  27. J Endocrinol. 2021 Aug 01. pii: JOE-21-0123.R1. [Epub ahead of print]
    Nicotinamide riboside has minimal impact on energy metabolism in mouse models of mild obesity.
    David M Cartwright, Lucy A Oakey, Rachel S Fletcher, Craig L Doig, Silke Heising, Dean P Larner, Daniela Nasteska, Caitlin E Berry, Sam R Heaselgrave, Christian Ludwig, David J Hodson, Gareth G Lavery, Antje Garten.
      Supplementation with precursors of nicotinamide adenine dinucleotide (NAD) has been shown to prevent and reverse insulin resistance, mitochondrial dysfunction and liver damage in mouse models of diet-induced obesity. We asked whether beneficial effects of supplementation with the NAD precursor nicotinamide riboside (NR) are dependent on mouse strain. We compared the effects of NR supplementation on whole-body energy metabolism and mitochondrial function in mildly obese C57BL/6N and C57BL/6J mice, two commonly used strains to investigate metabolism. Male C57BL/6N and C57BL/6J mice were fed a high-fat diet (HFD) or standard chow with or without NR supplementation for 8 weeks. Body and organ weights, glucose tolerance and metabolic parameters as well as mitochondrial O2 flux in liver and muscle fibers were assessed. We found that NR supplementation had no influence on body or organ weight, glucose metabolism or hepatic lipid accumulation, energy expenditure or metabolic flexibility, but increased mitochondrial respiration in soleus muscle in both mouse strains. Strain-dependent differences were detected for body and fat depot weight, fasting blood glucose, hepatic lipid accumulation and energy expenditure. We conclude that, in mild obesity, NR supplementation does not alter metabolic phenotype in two commonly used laboratory mouse strains.
    DOI:  https://doi.org/10.1530/JOE-21-0123
  28. Nat Struct Mol Biol. 2021 Aug;28(8): 662-670
    The tumor suppressor folliculin inhibits lactate dehydrogenase A and regulates the Warburg effect.
    Mark R Woodford, Alexander J Baker-Williams, Rebecca A Sager, Sarah J Backe, Adam R Blanden, Fiza Hashmi, Priyanka Kancherla, Alessandro Gori, David R Loiselle, Matteo Castelli, Stefano A Serapian, Giorgio Colombo, Timothy A Haystead, Sandra M Jensen, William G Stetler-Stevenson, Stewart N Loh, Laura S Schmidt, W Marston Linehan, Alaji Bah, Dimitra Bourboulia, Gennady Bratslavsky, Mehdi Mollapour.
      Aerobic glycolysis in cancer cells, also known as the 'Warburg effect', is driven by hyperactivity of lactate dehydrogenase A (LDHA). LDHA is thought to be a substrate-regulated enzyme, but it is unclear whether a dedicated intracellular protein also regulates its activity. Here, we identify the human tumor suppressor folliculin (FLCN) as a binding partner and uncompetitive inhibitor of LDHA. A flexible loop within the amino terminus of FLCN controls movement of the LDHA active-site loop, tightly regulating its enzyme activity and, consequently, metabolic homeostasis in normal cells. Cancer cells that experience the Warburg effect show FLCN dissociation from LDHA. Treatment of these cells with a decapeptide derived from the FLCN loop region causes cell death. Our data suggest that the glycolytic shift of cancer cells is the result of FLCN inactivation or dissociation from LDHA. Together, FLCN-mediated inhibition of LDHA provides a new paradigm for the regulation of glycolysis.
    DOI:  https://doi.org/10.1038/s41594-021-00633-2
  29. Cell Rep. 2021 Aug 10. pii: S2211-1247(21)00946-3. [Epub ahead of print]36(6): 109516
    NAD+ supplement potentiates tumor-killing function by rescuing defective TUB-mediated NAMPT transcription in tumor-infiltrated T cells.
    Yuetong Wang, Fei Wang, Lihua Wang, Shizhen Qiu, Yufeng Yao, Chenxu Yan, Xuexue Xiong, Xuyong Chen, Quanquan Ji, Jian Cao, Ganglong Gao, Dake Li, Liye Zhang, Zhiqian Guo, Ruoning Wang, Haopeng Wang, Gaofeng Fan.
      Although tumor-infiltrating lymphocytes (TILs) maintain their ability to proliferate, persist, and eradicate tumors, they are frequently dysfunctional in situ. By performing both whole-genome CRISPR and metabolic inhibitor screens, we identify that nicotinamide phosphoribosyltransferase (NAMPT) is required for T cell activation. NAMPT is low in TILs, and its expression is controlled by the transcriptional factor Tubby (TUB), whose activity depends on the T cell receptor-phospholipase C gamma (TCR-PLCγ) signaling axis. The intracellular level of NAD+, whose synthesis is dependent on the NAMPT-mediated salvage pathway, is also decreased in TILs. Liquid chromatography-mass spectrometry (LC-MS) and isotopic labeling studies confirm that NAD+ depletion led to suppressed glycolysis, disrupted mitochondrial function, and dampened ATP synthesis. Excitingly, both adoptive CAR-T and anti-PD1 immune checkpoint blockade mouse models demonstrate that NAD+ supplementation enhanced the tumor-killing efficacy of T cells. Collectively, this study reveals that an impaired TCR-TUB-NAMPT-NAD+ axis leads to T cell dysfunction in the tumor microenvironment, and an over-the-counter nutrient supplement of NAD+ could boost T-cell-based immunotherapy.
    Keywords:  CAR-T; NAD(+) supplement; NAMPT; PD-1; T cell activation; TUB; cancer immunotherapy
    DOI:  https://doi.org/10.1016/j.celrep.2021.109516
  30. J Biol Chem. 2021 Aug 07. pii: S0021-9258(21)00860-7. [Epub ahead of print] 101058
    The m6A methyltransferase METTL3 modifies PGC-1α mRNA promoting mitochondrial dysfunction and oxLDL-induced inflammation in monocytes.
    Xinning Zhang, Xin Li, Hongti Jia, Guoshun An, Juhua Ni.
      Mitochondrial biogenesis and energy metabolism are essential for regulating the inflammatory state of monocytes. This state is partially controlled by peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α), a coactivator that regulates mitochondrial biogenesis and energy metabolism. Disruption of these processes can also contribute to the initiation of chronic inflammatory diseases, such as pulmonary fibrosis, atherosclerosis and rheumatoid arthritis. Methyltransferase-like 3 (METTL3)-dependent N6-methyladenosine (m6A) methylation has recently been shown to regulate a variety of inflammatory processes. However, the role of m6A mRNA methylation in affecting mitochondrial metabolism in monocytes under inflammation is unclear, nor is there an established relationship between m6A methylation and PGC-1α. In this study, we identified a novel mechanism by which METTL3 acts during oxidized low-density lipoprotein (oxLDL)-induced monocyte inflammation, where METTL3 and YTH N6-methyladenosine RNA binding protein 2 (YTHDF2) cooperatively modify PGC-1α mRNA, mediating its degradation, decreasing PGC-1α protein levels, and thereby enhancing the inflammatory response. METTL3 coordinated with YTHDF2 to suppress the expression of PGC-1α, as well as that of cytochrome c (CYCS) and NADH:ubiquinone oxidoreductase subunit C2 (NDUFC2), and reduced ATP production and oxygen consumption rate (OCR). This subsequently increased the accumulation of cellular and mitochondrial reactive oxygen species (ROS) and the levels of proinflammatory cytokines in inflammatory monocytes. These data may provide new insights into the role of METTL3-dependent m6A modification of PGC-1α mRNA in the monocyte inflammation response. These data also contribute to a more comprehensive understanding of the pathogenesis of monocyte-macrophage inflammation-associated diseases, such as pulmonary fibrosis, atherosclerosis and rheumatoid arthritis.
    Keywords:  METTL3; PGC-1α; RNA methylation; inflammation; m(6)A modification; mitochondria; monocytes; post-transcriptional regulation
    DOI:  https://doi.org/10.1016/j.jbc.2021.101058
  31. Nat Commun. 2021 08 10. 12(1): 4814
    Two parallel pathways connect glutamine metabolism and mTORC1 activity to regulate glutamoptosis.
    Clément Bodineau, Mercedes Tomé, Sarah Courtois, Ana S H Costa, Marco Sciacovelli, Benoit Rousseau, Elodie Richard, Pierre Vacher, Carlos Parejo-Pérez, Emilie Bessede, Christine Varon, Pierre Soubeyran, Christian Frezza, Piedad Del Socorro Murdoch, Victor H Villar, Raúl V Durán.
      Glutamoptosis is the induction of apoptotic cell death as a consequence of the aberrant activation of glutaminolysis and mTORC1 signaling during nutritional imbalance in proliferating cells. The role of the bioenergetic sensor AMPK during glutamoptosis is not defined yet. Here, we show that AMPK reactivation blocks both the glutamine-dependent activation of mTORC1 and glutamoptosis in vitro and in vivo. We also show that glutamine is used for asparagine synthesis and the GABA shunt to produce ATP and to inhibit AMPK, independently of glutaminolysis. Overall, our results indicate that glutamine metabolism is connected with mTORC1 activation through two parallel pathways: an acute alpha-ketoglutarate-dependent pathway; and a secondary ATP/AMPK-dependent pathway. This dual metabolic connection between glutamine and mTORC1 must be considered for the future design of therapeutic strategies to prevent cell growth in diseases such as cancer.
    DOI:  https://doi.org/10.1038/s41467-021-25079-4
  32. iScience. 2021 Aug 20. 24(8): 102869
    Dissecting the concordant and disparate roles of NDUFAF3 and NDUFAF4 in mitochondrial complex I biogenesis.
    Anjaneyulu Murari, Shauna-Kay Rhooms, Christian Garcia, Tong Liu, Hong Li, Bibhuti Mishra, Cassie Deshong, Edward Owusu-Ansah.
      Distinct sub-assemblies (modules) of mitochondrial complex I (CI) are assembled with the assistance of CI Assembly Factors (CIAFs) through mechanisms that are incompletely defined. Here, using genetic analyses in Drosophila, we report that when either of the CIAFs - NDUFAF3 or NDUFAF4 - is disrupted, biogenesis of the Q-, N-, and PP-b-modules of CI is impaired. This is due, at least in part, to the compromised integration of NDUFS3 and NDUFS5 into the Q-, and PP-b-modules, respectively, coupled with a destabilization of another CIAF, TIMMDC1, in assembly intermediates. Notably, forced expression of NDUFAF4 rescues the biogenesis defects in the Q-module and some aspects of the defects in the PP-b-module of CI when NDUFAF3 is disrupted. Altogether, our studies furnish new fundamental insights into the mechanism by which NDUFAF3 and NDUFAF4 regulate CI assembly and raises the possibility that certain point mutations in NDUFAF3 may be rescued by overexpression of NDUFAF4.
    Keywords:  metabolic engineering; molecular genetics; molecular mechanism of gene regulation
    DOI:  https://doi.org/10.1016/j.isci.2021.102869
  33. Nature. 2021 Aug 11.
    Cycling cancer persister cells arise from lineages with distinct programs.
    Yaara Oren, Michael Tsabar, Michael S Cuoco, Liat Amir-Zilberstein, Heidie F Cabanos, Jan-Christian Hütter, Bomiao Hu, Pratiksha I Thakore, Marcin Tabaka, Charles P Fulco, William Colgan, Brandon M Cuevas, Sara A Hurvitz, Dennis J Slamon, Amy Deik, Kerry A Pierce, Clary Clish, Aaron N Hata, Elma Zaganjor, Galit Lahav, Katerina Politi, Joan S Brugge, Aviv Regev.
      Non-genetic mechanisms have recently emerged as important drivers of cancer therapy failure1, where some cancer cells can enter a reversible drug-tolerant persister state in response to treatment2. Although most cancer persisters remain arrested in the presence of the drug, a rare subset can re-enter the cell cycle under constitutive drug treatment. Little is known about the non-genetic mechanisms that enable cancer persisters to maintain proliferative capacity in the presence of drugs. To study this rare, transiently resistant, proliferative persister population, we developed Watermelon, a high-complexity expressed barcode lentiviral library for simultaneous tracing of each cell's clonal origin and proliferative and transcriptional states. Here we show that cycling and non-cycling persisters arise from different cell lineages with distinct transcriptional and metabolic programs. Upregulation of antioxidant gene programs and a metabolic shift to fatty acid oxidation are associated with persister proliferative capacity across multiple cancer types. Impeding oxidative stress or metabolic reprogramming alters the fraction of cycling persisters. In human tumours, programs associated with cycling persisters are induced in minimal residual disease in response to multiple targeted therapies. The Watermelon system enabled the identification of rare persister lineages that are preferentially poised to proliferate under drug pressure, thus exposing new vulnerabilities that can be targeted to delay or even prevent disease recurrence.
    DOI:  https://doi.org/10.1038/s41586-021-03796-6
  34. Front Oncol. 2021 ;11 715593
    ME2 Promotes Proneural-Mesenchymal Transition and Lipogenesis in Glioblastoma.
    Mengting Yang, Xi Chen, Junyao Zhang, Ermeng Xiong, Qianqian Wang, Wenjing Fang, Li Li, Fei Fei, Aihua Gong.
      Malic enzyme 2 (ME2) catalyzes the formation of pyruvate from malic acid and is abnormally expressed in some tumors. However, the exact effects of ME2 on proneural-mesenchymal transition (PMT) and lipogenesis in glioblastoma multiforme (GBM) remain unexplored. Here, we found that ME2 expression was significantly higher in GBM than in normal brain tissues and negatively correlated with overall survival of patients with GBM. Furthermore, we demonstrated that ME2 was positively correlated with mesenchymal features in GBM and promoted proliferation, migration, and invasion of glioma cells. Moreover, ME2 upregulated the expression of mesenchymal markers (N-cadherin, vimentin, YKL40, and MET), whereas it inhibited the expression of proneural maker OLIG2, indicating that ME2 might promote PMT in GBM. We also found that ME2 inhibited the production of mitochondrial reactive oxygen species and AMPK phosphorylation, resulting in SREBP-1 maturation and nuclear localization and enhancing the ACSS2 lipogenesis pathway. Taken together, these results suggest that ME2 promotes PMT and is linked with reprogramming of lipogenesis via AMPK-SREBP-1-ACSS2 signaling in GBM. Therefore, ME2 has potential as a new classification marker in GBM and could provide a new approach to glioma treatment.
    Keywords:  AMPK; glioblastoma; lipogenesis; malic enzyme 2; proneural–mesenchymal transition
    DOI:  https://doi.org/10.3389/fonc.2021.715593
  35. Nat Commun. 2021 08 12. 12(1): 4878
    Targeting of eIF6-driven translation induces a metabolic rewiring that reduces NAFLD and the consequent evolution to hepatocellular carcinoma.
    Alessandra Scagliola, Annarita Miluzio, Gabriele Ventura, Stefania Oliveto, Chiara Cordiglieri, Nicola Manfrini, Delia Cirino, Sara Ricciardi, Luca Valenti, Guido Baselli, Roberta D'Ambrosio, Marco Maggioni, Daniela Brina, Alberto Bresciani, Stefano Biffo.
      A postprandial increase of translation mediated by eukaryotic Initiation Factor 6 (eIF6) occurs in the liver. Its contribution to steatosis and disease is unknown. In this study we address whether eIF6-driven translation contributes to disease progression. eIF6 levels increase throughout the progression from Non-Alcoholic Fatty Liver Disease (NAFLD) to hepatocellular carcinoma. Reduction of eIF6 levels protects the liver from disease progression. eIF6 depletion blunts lipid accumulation, increases fatty acid oxidation (FAO) and reduces oncogenic transformation in vitro. In addition, eIF6 depletion delays the progression from NAFLD to hepatocellular carcinoma, in vivo. Mechanistically, eIF6 depletion reduces the translation of transcription factor C/EBPβ, leading to a drop in biomarkers associated with NAFLD progression to hepatocellular carcinoma and preserves mitochondrial respiration due to the maintenance of an alternative mTORC1-eIF4F translational branch that increases the expression of transcription factor YY1. We provide proof-of-concept that in vitro pharmacological inhibition of eIF6 activity recapitulates the protective effects of eIF6 depletion. We hypothesize the existence of a targetable, evolutionarily conserved translation circuit optimized for lipid accumulation and tumor progression.
    DOI:  https://doi.org/10.1038/s41467-021-25195-1
  36. Acta Pharm Sin B. 2021 Jul;11(7): 1853-1866
    Pharmacologically targeting molecular motor promotes mitochondrial fission for anti-cancer.
    Yi Qian, Meimei Zhao, Qinghua Han, Jingkang Wang, Lixi Liao, Heng Yang, Dan Liu, Pengfei Tu, Hong Liang, Kewu Zeng.
      Mitochondrial shape rapidly changes by dynamic balance of fusion and fission to adjust to constantly changing energy demands of cancer cells. Mitochondrial dynamics balance is exactly regulated by molecular motor consisted of myosin and actin cytoskeleton proteins. Thus, targeting myosin-actin molecular motor is considered as a promising strategy for anti-cancer. In this study, we performed a proof-of-concept study with a natural-derived small-molecule J13 to test the feasibility of anti-cancer therapeutics via pharmacologically targeting molecular motor. Here, we found J13 could directly target myosin-9 (MYH9)-actin molecular motor to promote mitochondrial fission progression, and markedly inhibited cancer cells survival, proliferation and migration. Mechanism study revealed that J13 impaired MYH9-actin interaction to inactivate molecular motor, and caused a cytoskeleton-dependent mitochondrial dynamics imbalance. Moreover, stable isotope labeling with amino acids in cell culture (SILAC) technology-coupled with pulldown analysis identified HSPA9 as a crucial adaptor protein connecting MYH9-actin molecular motor to mitochondrial fission. Taken together, we reported the first natural small-molecule directly targeting MYH9-actin molecular motor for anti-cancer translational research. Besides, our study also proved the conceptual practicability of pharmacologically disrupting mitochondrial fission/fusion dynamics in human cancer therapy.
    Keywords:  Anti-cancer; CAM, chick embryo chorioallantoic membrane; CETSA, cellular thermal shift assay; Co-IP, co-immunoprecipitation; DAPI, 4′,6-diamidino-2-phenylindole; ER, endoplasmic reticulum; HE, hematoxylin–eosin staining; HSPA9; HSPA9, heat-shock protein A9; HUVEC, human umbilical vein endothelial cells; IHC, immunohistochemistry; LIHC, liver hepatocellular carcinoma; Liver hepatocellular carcinoma; MMP, mitochondrial membrane potential; MYH9; MYH9, myosin-9; Mitochondrial fission; Molecular motor; SILAC, stable isotope labeling with amino acids in cell culture; SPR, surface plasmon resonance; Small molecule; TUNEL, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling; Target identification
    DOI:  https://doi.org/10.1016/j.apsb.2021.01.011
  37. Exp Cell Res. 2021 Aug 09. pii: S0014-4827(21)00331-1. [Epub ahead of print] 112778
    Heterogeneous response of cancer-associated fibroblasts to the glucose deprivation through mitochondrial calcium uniporter.
    Sung-Hyun Hwang, Yeseul Yang, Jae-Ha Jung, Yongbaek Kim.
      Cancer-associated fibroblasts (CAFs) are an abundant component of the tumor microenvironment and have distinct features from normal fibroblasts (NFs). However, the discriminative nature of heterogeneous CAFs under glucose starvation remains unknown. In this study, we investigated the changes in the mitochondrial calcium concentration and relevant intracellular machinery in CAFs under glucose-deficient conditions. Xenografted tumor masses were dissected into multiple pieces and subjected to the CAF isolation using magnetically activated cell sorting (MACS). NFs were separated from the normal lung and skin. Under glucose starvation, CAFs from the tumor mass exhibited heterogeneity in cell proliferation, ATP production and calcium concentration. Compared to NFs, mitochondrial calcium concentration was significantly higher in glucose-starved CAFs with upregulation of mitochondrial calcium uniporter (MCU) that led to enhancement of ATP production and cell growth. Intriguingly, treatment of glucose-starved CAFs with oligomycin increased apoptosis by disrupted calcium homeostasis following overactivation of the mPTP. Moreover, oligomycin-induced apoptosis was mitigated by calcium chelation. This study demonstrated that the discriminative calcium influx to mitochondria through MCU coordinated cell growth and apoptosis in glucose-starved CAFs but not in NFs.
    Keywords:  Cancer-associated fibroblasts; Glucose-deficient condition; Mitochondrial calcium; Mitochondrial calcium uniporter; TGF-β
    DOI:  https://doi.org/10.1016/j.yexcr.2021.112778
  38. Cancer Discov. 2020 Feb;10(2): 173
    An SLC1A5 Variant Transports Glutamine to Mitochondria in Cancer Cells.
      A variant of SLC1A5 (SLC1A5_var) fueled pancreatic cancer cells' mitochondria with glutamine.
    DOI:  https://doi.org/10.1158/2159-8290.CD-RW2020-003
  39. Dig Dis Sci. 2021 Aug 09.
    Colorectal Cancer Cell Differentiation Is Dependent on the Repression of Aerobic Glycolysis by NDRG2-TXNIP Axis.
    Junbi Hu, Lin Feng, Mudan Ren, Yan Zhao, Guifang Lu, Xinlan Lu, Yarui Li, Xin Wang, Xin Bu, Shuai Wang, Liangliang Shen, Shuixiang He.
       BACKGROUND: Poorly differentiated colorectal cancers are more aggressive. Metabolism reprogramming is a significant hallmark in cancer, and aerobic glycolysis is common. However, how cancer cells reprogramming glucose metabolism contributes to cell differentiation was largely unknown. Previous studies have reported that tumor suppressor NDRG2 could promote colorectal cancers differentiation.
    AIMS: This study aims to demonstrate that NDRG2 promotes the differentiation of colorectal cancers, potentially through the inhibition of aerobic glycolysis via TXNIP induction.
    METHODS: Western blotting, qRT-PCR and immunohistochemical staining were used to detect the expression of related molecules. MTT assay was used to reflect cell viability and proliferation. Immunofluorescent assay was performed to identify the expression and distribution of molecules. Luciferase analysis and CHIP assays were used to investigate the mechanism. Bioinformatic analysis was performed to predict the relevance.
    RESULTS: In colorectal cancers, NDRG2 could inhibit cell proliferation, reduce glucose uptake and decrease expression of key glycolysis enzymes. Upregulated NDRG2 is associated with differentiated cancer. However, deletion of TXNIP, a classic glucose metabolism inhibitor, could obviously alter the function of NDRG2 in differentiation, glucose uptake, expression of key glycolysis enzymes and proliferation. Mechanistically, high glucose flux promotes the activity of TXNIP promoter. And NDRG2 promotes the occupancy of transcription factor Mondo A on TXNIP promoter, predominantly through the suppression of c-myc, which could complete with Mondo A binding to TXNIP promoter. In clinical samples, high expression of TXNIP indicates good prognosis and outcome.
    CONCLUSIONS: NDRG2-dependent induction of TXNIP is critical for the aerobic glycolysis during colorectal cancers differentiation.
    Keywords:  Colorectal cancer; Differentiation; Glycolysis; NDRG2; TXNIP
    DOI:  https://doi.org/10.1007/s10620-021-07188-8
  40. Front Cell Dev Biol. 2021 ;9 720490
    Metabolism and Innate Immunity Meet at the Mitochondria.
    Amir Bahat, Thomas MacVicar, Thomas Langer.
      Mitochondria are master regulators of metabolism and have emerged as key signalling organelles of the innate immune system. Each mitochondrion harbours potent agonists of inflammation, including mitochondrial DNA (mtDNA), which are normally shielded from the rest of the cell and extracellular environment and therefore do not elicit detrimental inflammatory cascades. Mitochondrial damage and dysfunction can lead to the cytosolic and extracellular exposure of mtDNA, which triggers inflammation in a number of diseases including autoimmune neurodegenerative disorders. However, recent research has revealed that the extra-mitochondrial exposure of mtDNA is not solely a negative consequence of mitochondrial damage and pointed to an active role of mitochondria in innate immunity. Metabolic cues including nucleotide imbalance can stimulate the release of mtDNA from mitochondria in order to drive a type I interferon response. Moreover, important effectors of the innate immune response to pathogen infection, such as the mitochondrial antiviral signalling protein (MAVS), are located at the mitochondrial surface and modulated by the cellular metabolic status and mitochondrial dynamics. In this review, we explore how and why metabolism and innate immunity converge at the mitochondria and describe how mitochondria orchestrate innate immune signalling pathways in different metabolic scenarios. Understanding how cellular metabolism and metabolic programming of mitochondria are translated into innate immune responses bears relevance to a broad range of human diseases including cancer.
    Keywords:  CGAS; MAVS; STING; innate immunity; metabolism; mitochondria; mitochondrial DNA
    DOI:  https://doi.org/10.3389/fcell.2021.720490
  41. Cancer Res. 2021 Aug 12. pii: canres.0567.2021. [Epub ahead of print]
    Stem cell factor SOX2 confers ferroptosis resistance in lung cancer via upregulation of SLC7A11.
    Xinbo Wang, Yueqing Chen, Xudong Wang, Hongling Tian, Yanjin Wang, Jiali Jin, Zezhi Shan, Yu'e Liu, Zhenyu Cai, Xinyuan Tong, Yi Luan, Xiao Tan, Bing Luan, Xin Ge, Hongbin Ji, Xuejun Jiang, Ping Wang.
      Ferroptosis is a lipid peroxidation-dependent cell death caused by metabolic dysfunction. Ferroptosis-associated enzymes are promising therapeutic targets for cancer treatment. However, such therapeutic strategies show limited efficacy due to drug resistance and other largely unknown underlying mechanisms. Here we report that cystine transporter SLC7A11 is upregulated in lung cancer stem-like cells (CSLC) and can be activated by stem cell transcriptional factor SOX2. Mutation of SOX2 binding site in SLC7A11 promoter reduced SLC7A11 expression and increased sensitivity to ferroptosis in cancer cells. Oxidation at Cys265 of SOX2 inhibited its activity and decreased the self-renewal capacity of CSLCs. Moreover, tumors with high SOX2 expression were more resistant to ferroptosis, and SLC7A11 expression was positively correlated with SOX2 in both mouse and human lung cancer tissue. Together, our study provides a mechanism by which cancer cells evade ferroptosis and suggests that oxidation of SOX2 can be a potential therapeutic target for cancer treatment.
    DOI:  https://doi.org/10.1158/0008-5472.CAN-21-0567
  42. Adv Healthc Mater. 2021 Aug 13. e2100978
    Mitochondrial Metabolism Targeted Nanoplatform for Efficient Triple-Negative Breast Cancer Combination Therapy.
    Lu Lu, Genhua Liu, Chuanchuan Lin, Ke Li, Tingting He, Jixi Zhang, Zhong Luo, Kaiyong Cai.
      Tumor reprogram pathway of mitochondrial metabolism is an emerging approach for malignant tumor treatment, such as triple-negative breast cancer. In this study, a tumor/mitochondria cascaded targeting, adenosine-triphosphate (ATP) responsive nanocarrier of zeolitic imidazolate framework-90 (ZIF-90) for breast cancer combination therapy is reported. Atovaquone (AVO) and hemin are loaded into ZIF-90, then a peptide iRGD with tumor-targeting ability is modified on the ZIF-90 nanoplatform. Hemin can specifically degrade BTB and CNC homology1 (BACH1), resulting in the changes of mitochondrial metabolism, and AVO acts as the inhibitor of the electron transport chain (ETC). The degradation of BACH1 using hemin can effectively improve the anti-tumor efficiency of mitochondrial metabolism inhibitor AVO, by increasing dependency on mitochondrial respiration. This nanoplatform displays both tumor-targeting and mitochondria-targeting capacity with high level of ATP responsive drug release behavior. The specific characteristic of mitochondria-targeting ability of this nanoplatform can increase the accumulation of AVO in the mitochondria, and in turn, can effectively improve the inhibition of the ETC. Both in vitro and in vivo results reveal that this composite nanocarrier has excellent tumor inhibition ability with limited side effects. Accordingly, this study provides an attractive strategy in the mitochondrial metabolism for cancer targeted therapy.
    Keywords:  BACH1; cascaded targeting; electron transport chains; mitochondrial metabolism; zeolitic imidazolate framework-90
    DOI:  https://doi.org/10.1002/adhm.202100978
  43. Nat Commun. 2021 08 12. 12(1): 4905
    Spontaneous hydrolysis and spurious metabolic properties of α-ketoglutarate esters.
    Seth J Parker, Joel Encarnación-Rosado, Kate E R Hollinshead, David M Hollinshead, Leonard J Ash, Juan A K Rossi, Elaine Y Lin, Albert S W Sohn, Mark R Philips, Drew R Jones, Alec C Kimmelman.
      α-ketoglutarate (KG), also referred to as 2-oxoglutarate, is a key intermediate of cellular metabolism with pleiotropic functions. Cell-permeable esterified analogs are widely used to study how KG fuels bioenergetic and amino acid metabolism and DNA, RNA, and protein hydroxylation reactions, as cellular membranes are thought to be impermeable to KG. Here we show that esterified KG analogs rapidly hydrolyze in aqueous media, yielding KG that, in contrast to prevailing assumptions, imports into many cell lines. Esterified KG analogs exhibit spurious KG-independent effects on cellular metabolism, including extracellular acidification, arising from rapid hydrolysis and de-protonation of α-ketoesters, and significant analog-specific inhibitory effects on glycolysis or mitochondrial respiration. We observe that imported KG decarboxylates to succinate in the cytosol and contributes minimally to mitochondrial metabolism in many cell lines cultured in normal conditions. These findings demonstrate that nuclear and cytosolic KG-dependent reactions may derive KG from functionally distinct subcellular pools and sources.
    DOI:  https://doi.org/10.1038/s41467-021-25228-9
  44. Biochem Biophys Res Commun. 2021 Aug 06. pii: S0006-291X(21)01149-9. [Epub ahead of print]573 1-8
    ATXN7L3B promotes hepatocellular carcinoma stemness and is downregulated by metformin.
    Bi Chen, Jong-Ho Cha, Meisi Yan, Nengqi Cao, Peng Ye, Xiuwen Yan, Wen-Hao Yang.
      Hepatocellular carcinoma (HCC) is the major cause of liver cancer-associated morality. Metformin, used for treating type 2 diabetes, has antitumor activity and reduces the risk of some diabetes-related tumors, such as liver and breast cancer. However, the mechanisms underlying metformin's effects in HCC remain unclear. To identify genes associated with metformin treatment in HCC, we conducted transcriptomic and proteomic analyses in HCC cells treated with or without metformin. We identified 41 differentially expressed genes upon metformin treatment. Among them, Ataxin 7 Like 3B (ATXN7L3B), which is a negative regulator of the Spt-Ada-Gcn5 acetyltransferase (SAGA) deubiquitinase (DUB) module and has relatively unknown functions in cancer, attracted our attention. We observed that metformin reduced ATXN7L3B level in HCC cells. ATXN7L3B expression was significantly negatively correlated with survival in liver cancer patients. We also demonstrated that ATXN7L3B promoted HCC stemness. Metformin treatment decreased ATXN7L3B-induced tumor-initiating ability in a HCC mouse model, implying that metformin may inhibit cancer stemness by downregulating ATXN7L3B. Our study supports the antitumor activity of metformin and its potential as an anticancer drug for HCC treatment.
    Keywords:  ATXN7L3B; Cancer stemness; Hepatocellular carcinoma; Metformin
    DOI:  https://doi.org/10.1016/j.bbrc.2021.08.009
  45. Am J Physiol Endocrinol Metab. 2021 08 09.
    Mito-Mendelian Interactions Alter in vivo Glucose Metabolism and Insulin Sensitivity in Healthy Mice.
    Melissa June Sammy, Ashley W Connelly, Jamelle A Brown, Cassie Holleman, Kirk M Habegger, Scott W Ballinger.
      The regulation of euglycemia is essential for human health with both chronic hypoglycemia and hyperglycemia having detrimental effects. Diabetes risk increases with age and exhibits racial disparity. Interestingly, mitochondrial DNA (mtDNA) damage accumulates with age and its sequence varies with geographic maternal origins (maternal race). From these two observations, we hypothesized that mtDNA background may contribute to glucose metabolism and insulin sensitivity. Pronuclear transfer was used to generate Mitochondrial-Nuclear eXchange (MNX) mice to directly test this hypothesis, by assessing physiologic parameters of glucose metabolism in nuclear isogenic C57BL/6J mice harboring either a C57BL/6J (C57n:C57mt wild-type - control) or C3H/HeN mtDNA (C57n:C3Hmt - MNX). All mice were fed normal chow diets. MNX mice were significantly leaner, had lower leptin levels and were more insulin sensitive, with lower modified Homeostatic Model Assessment of Insulin Resistance (mHOMA-IR) values and enhanced insulin action when compared to their control counterparts. Further interrogation of muscle insulin signaling revealed higher phosphorylated Akt/total Akt ratios in MNX animals relative to control, consistent with greater insulin sensitivity. Overall, these results are consistent with the hypothesis that different mtDNA combinations on the same nuclear DNA (nDNA) background can significantly impact glucose metabolism and insulin sensitivity in healthy mice.
    Keywords:  Mitochondrial Nuclear eXchange; adiposity; glucose metabolism; insulin sensitivity; mitochondrial DNA
    DOI:  https://doi.org/10.1152/ajpendo.00069.2021
  46. Oxid Med Cell Longev. 2021 ;2021 6697861
    Intercellular Transfer of Mitochondria between Senescent Cells through Cytoskeleton-Supported Intercellular Bridges Requires mTOR and CDC42 Signalling.
    Hannah E Walters, Lynne S Cox.
      Cellular senescence is a state of irreversible cell proliferation arrest induced by various stressors including telomere attrition, DNA damage, and oncogene induction. While beneficial as an acute response to stress, the accumulation of senescent cells with increasing age is thought to contribute adversely to the development of cancer and a number of other age-related diseases, including neurodegenerative diseases for which there are currently no effective disease-modifying therapies. Non-cell-autonomous effects of senescent cells have been suggested to arise through the SASP, a wide variety of proinflammatory cytokines, chemokines, and exosomes secreted by senescent cells. Here, we report an additional means of cell communication utilised by senescent cells via large numbers of membrane-bound intercellular bridges-or tunnelling nanotubes (TNTs)-containing the cytoskeletal components actin and tubulin, which form direct physical connections between cells. We observe the presence of mitochondria in these TNTs and show organelle transfer through the TNTs to adjacent cells. While transport of individual mitochondria along single TNTs appears by time-lapse studies to be unidirectional, we show by differentially labelled co-culture experiments that organelle transfer through TNTs can occur between different cells of equivalent cell age, but that senescent cells, rather than proliferating cells, appear to be predominant mitochondrial donors. Using small molecule inhibitors, we demonstrate that senescent cell TNTs are dependent on signalling through the mTOR pathway, which we further show is mediated at least in part through the downstream actin-cytoskeleton regulatory factor CDC42. These findings have significant implications for the development of senomodifying therapies, as they highlight the need to account for local direct cell-cell contacts as well as the SASP in order to treat cancer and diseases of ageing in which senescence is a key factor.
    DOI:  https://doi.org/10.1155/2021/6697861
  47. Mol Cell. 2021 Jul 30. pii: S1097-2765(21)00590-6. [Epub ahead of print]
    A proteomic and phosphoproteomic landscape of KRAS mutant cancers identifies combination therapies.
    Zhiwei Liu, Yingluo Liu, Lili Qian, Shangwen Jiang, Xiameng Gai, Shu Ye, Yuehong Chen, Xiaomin Wang, Linhui Zhai, Jun Xu, Congying Pu, Jing Li, Fuchu He, Min Huang, Minjia Tan.
      KRAS mutant cancer, characterized by the activation of a plethora of phosphorylation signaling pathways, remains a major challenge for cancer therapy. Despite recent advancements, a comprehensive profile of the proteome and phosphoproteome is lacking. This study provides a proteomic and phosphoproteomic landscape of 43 KRAS mutant cancer cell lines across different tissue origins. By integrating transcriptomics, proteomics, and phosphoproteomics, we identify three subsets with distinct biological, clinical, and therapeutic characteristics. The integrative analysis of phosphoproteome and drug sensitivity information facilitates the identification of a set of drug combinations with therapeutic potentials. Among them, we demonstrate that the combination of DOT1L and SHP2 inhibitors is an effective treatment specific for subset 2 of KRAS mutant cancers, corresponding to a set of TCGA clinical tumors with the poorest prognosis. Together, this study provides a resource to better understand KRAS mutant cancer heterogeneity and identify new therapeutic possibilities.
    Keywords:  DOT1L; KRAS mutation; SHP2; cancer; drug sensitivity; heterogeneity; phosphoproteomics; proteomics; subtype; therapy
    DOI:  https://doi.org/10.1016/j.molcel.2021.07.021
  48. Nat Commun. 2021 08 09. 12(1): 4787
    IceR improves proteome coverage and data completeness in global and single-cell proteomics.
    Mathias Kalxdorf, Torsten Müller, Oliver Stegle, Jeroen Krijgsveld.
      Label-free proteomics by data-dependent acquisition enables the unbiased quantification of thousands of proteins, however it notoriously suffers from high rates of missing values, thus prohibiting consistent protein quantification across large sample cohorts. To solve this, we here present IceR (Ion current extraction Re-quantification), an efficient and user-friendly quantification workflow that combines high identification rates of data-dependent acquisition with low missing value rates similar to data-independent acquisition. Specifically, IceR uses ion current information for a hybrid peptide identification propagation approach with superior quantification precision, accuracy, reliability and data completeness compared to other quantitative workflows. Applied to plasma and single-cell proteomics data, IceR enhanced the number of reliably quantified proteins, improved discriminability between single-cell populations, and allowed reconstruction of a developmental trajectory. IceR will be useful to improve performance of large scale global as well as low-input proteomics applications, facilitated by its availability as an easy-to-use R-package.
    DOI:  https://doi.org/10.1038/s41467-021-25077-6
  49. Cancer Sci. 2021 Aug 07.
    The survival and proliferation of osteosarcoma cells are dependent on the mitochondrial BIG3-PHB2 complex formation.
    Shunichi Toki, Tetsuro Yoshimaru, Yosuke Matsushita, Hitoshi Aihara, Masaya Ono, Koichi Tsuneyama, Koichi Sairyo, Toyomasa Katagiri.
      Previous studies reported the critical role of the brefeldin A-inhibited guanine nucleotide exchange protein 3-prohibitin 2 (BIG3-PHB2) complex in modulating estrogen signaling activation in breast cancer cells, yet its pathophysiological roles in osteosarcoma (OS) cells remain elusive. Here, we report a novel function of BIG3-PHB2 in OS malignancy. BIG3-PHB2 complexes were localized mainly in mitochondria in OS cells, unlike in estrogen-dependent breast cancer cells. Depletion of endogenous BIG3 expression by small interfering RNA (siRNA) treatment led to significant inhibition of OS cell growth. Disruption of BIG3-PHB2 complex formation by treatment with specific peptide inhibitor also resulted in significant dose-dependent suppression of OS cell growth, migration, and invasion resulting from G2/M-phase arrest and in PARP cleavage, ultimately leading to PARP-1/AIF pathway activation-dependent apoptosis in OS cells. Subsequent proteomic and bioinformatic pathway analyses revealed that disruption of the BIG3-PHB2 complex might lead to downregulation of inner mitochondrial membrane protein complex activity. Our findings indicate that the mitochondrial BIG3-PHB2 complex might regulate PARP-1/AIF pathway-dependent apoptosis during OS cell proliferation and progression and that disruption of this complex may be a promising therapeutic strategy for OS.
    Keywords:  BIG3; PHB2; mitochondria; osteosarcoma; peptide inhibitor
    DOI:  https://doi.org/10.1111/cas.15099
  50. Nat Commun. 2021 Aug 13. 12(1): 4921
    Interacting evolutionary pressures drive mutation dynamics and health outcomes in aging blood.
    Kimberly Skead, Armande Ang Houle, Sagi Abelson, Mawusse Agbessi, Vanessa Bruat, Boxi Lin, David Soave, Liran Shlush, Stephen Wright, John Dick, Quaid Morris, Philip Awadalla.
      Age-related clonal hematopoiesis (ARCH) is characterized by age-associated accumulation of somatic mutations in hematopoietic stem cells (HSCs) or their pluripotent descendants. HSCs harboring driver mutations will be positively selected and cells carrying these mutations will rise in frequency. While ARCH is a known risk factor for blood malignancies, such as Acute Myeloid Leukemia (AML), why some people who harbor ARCH driver mutations do not progress to AML remains unclear. Here, we model the interaction of positive and negative selection in deeply sequenced blood samples from individuals who subsequently progressed to AML, compared to healthy controls, using deep learning and population genetics. Our modeling allows us to discriminate amongst evolutionary classes with high accuracy and captures signatures of purifying selection in most individuals. Purifying selection, acting on benign or mildly damaging passenger mutations, appears to play a critical role in preventing disease-predisposing clones from rising to dominance and is associated with longer disease-free survival. Through exploring a range of evolutionary models, we show how different classes of selection shape clonal dynamics and health outcomes thus enabling us to better identify individuals at a high risk of malignancy.
    DOI:  https://doi.org/10.1038/s41467-021-25172-8
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