bims-mibica Biomed News
on Mitochondrial bioenergetics in cancer
Issue of 2021‒05‒30
48 papers selected by
Kelsey Fisher-Wellman, East Carolina University
  1. Ethyl Isopropyl Amiloride Decreases Oxidative Phosphorylation and Increases Mitochondrial Fusion in Clonal Untransformed and Cancer Cells.
  2. Mass-spectrometry-based proteomics reveals mitochondrial supercomplexome plasticity.
  3. Generation of somatic mitochondrial DNA-replaced cells for mitochondrial dysfunction treatment.
  4. Excess Exogenous Pyruvate Inhibits Lactate Dehydrogenase Activity in Live Cells in a MCT1-Dependent Manner.
  5. ECSIT is a critical limiting factor for cardiac function.
  6. Preserved skeletal muscle oxidative capacity in older adults despite decreased cardiorespiratory fitness with aging.
  7. Mitochondrial localization and moderated activity are key to murine erythroid enucleation.
  8. Medium-chain acyl CoA dehydrogenase protects mitochondria from lipid peroxidation in glioblastoma.
  9. Genome-wide CRISPRi screening identifies OCIAD1 as a prohibitin client and regulatory determinant of mitochondrial Complex III assembly in human cells.
  10. Sulfide catabolism ameliorates hypoxic brain injury.
  11. Mitochondrial DNA variation and cancer.
  12. Mammary epithelial cells have lineage-rooted metabolic identities.
  13. Elevated type I interferon responses potentiate metabolic dysfunction, inflammation, and accelerated aging in mtDNA mutator mice.
  14. Calibration and measurement of mitochondrial pH in intact adult rat cardiomyocytes.
  15. NADPH supply and the contribution of NAD(P)+ transhydrogenase (NNT) to H2O2 balance in skeletal muscle mitochondria.
  16. Chromosomal copy number heterogeneity predicts survival rates across cancers.
  17. SIRT5 IS A DRUGGABLE METABOLIC VULNERABILITY IN ACUTE MYELOID LEUKEMIA.
  18. Metabolic reprogramming of terminally exhausted CD8+ T cells by IL-10 enhances anti-tumor immunity.
  19. Fatty acid β-oxidation promotes breast cancer stemness and metastasis via the miRNA-328-3p-CPT1A pathway.
  20. Iron supplementation regulates the progression of high fat diet induced obesity and hepatic steatosis via mitochondrial signaling pathways.
  21. The Functions, Methods, and Mobility of Mitochondrial Transfer Between Cells.
  22. Pan-cancer characterization of metabolism-related biomarkers identifies potential therapeutic targets.
  23. Mitochondrial efflux of citrate and isocitrate is fully dispensable for glucose-stimulated insulin secretion and pancreatic islet β-cell function.
  24. Structures of the human LONP1 protease reveal regulatory steps involved in protease activation.
  25. Chronic cold exposure induces autophagy to promote fatty acid oxidation, mitochondrial turnover, and thermogenesis in brown adipose tissue.
  26. ND‑09 inhibits chronic myeloid leukemia K562 cell growth by regulating BCR‑ABL signaling.
  27. Targeting NF-κB-dependent alkaliptosis for the treatment of venetoclax-resistant acute myeloid leukemia cells.
  28. pH-activated, mitochondria-targeted, and redox-responsive delivery of paclitaxel nanomicelles to overcome drug resistance and suppress metastasis in lung cancer.
  29. High doses of butyrate induce a reversible body temperature drop through transient proton leak in mitochondria of brain neurons.
  30. Single channel properties of mitochondrial large conductance potassium channel formed by BK-VEDEC splice variant.
  31. N6-Methyladenosine on mRNA facilitates a phase-separated nuclear body that suppresses myeloid leukemic differentiation.
  32. CaMKII oxidation is a critical performance/disease trade-off acquired at the dawn of vertebrate evolution.
  33. Mitochondrial transplantation therapy inhibit carbon tetrachloride-induced liver injury through scavenging free radicals and protecting hepatocytes.
  34. Extracellular vesicles activate ATM-Chk2 signaling pathway through the intercellular transfer of mitochondrial DNA in HBV-infected human hepatocytes.
  35. Mitochondrial ROS signals prevent excessive immune response.
  36. Restoration of energy homeostasis by SIRT6 extends healthy lifespan.
  37. Insulin action, type 2 diabetes, and branched-chain amino acids: a two-way street.
  38. Epigenetic induction of mitochondrial fission is required for maintenance of liver cancer initiating cells.
  39. VCP relocalization limits mitochondrial activity, GSH depletion, and ferroptosis during starvation in PC3 prostate cancer cells.
  40. Mitocytosis, a migrasome-mediated mitochondrial quality-control process.
  41. Single-cell metabolic imaging reveals a SLC2A3-dependent glycolytic burst in motile endothelial cells.
  42. A Quinol Anion as Catalytic Intermediate Coupling Proton Translocation With Electron Transfer in E. coli Respiratory Complex I.
  43. Mitochondrial targeted meganuclease as a platform to eliminate mutant mtDNA in vivo.
  44. The mitochondrial fission factor FIS1 promotes stemness of human lung cancer stem cells via mitophagy.
  45. Mitohormesis reprogrammes macrophage metabolism to enforce tolerance.
  46. Small molecule inhibitors of the mitochondrial ClpXP protease possess cytostatic potential and re-sensitize chemo-resistant cancers.
  47. Hexose-6-phosphate dehydrogenase blockade reverses prostate cancer drug resistance in xenograft models by glucocorticoid inactivation.
  48. Perm1 promotes cardiomyocyte mitochondrial biogenesis and protects against hypoxia/reoxygenation-induced damage in mice.
  1. Am J Physiol Cell Physiol. 2021 05 26.
    Ethyl Isopropyl Amiloride Decreases Oxidative Phosphorylation and Increases Mitochondrial Fusion in Clonal Untransformed and Cancer Cells.
    Sagar S Manoli, Kyle Kisor, Bradley A Webb, Diane L Barber.
      Many cancer cells, regardless of their tissue origin or genetic landscape, have increased expression or activity of the plasma membrane Na-H exchanger NHE1 and a higher intracellular pH (pHi) compared with untransformed cells. A current perspective that remains to be validated is that increased NHE1 activity and pHi enable a Warburg-like metabolic reprogramming of increased glycolysis and decreased mitochondrial oxidative phosphorylation. We tested this perspective and find it is not accurate for clonal pancreatic and breast cancer cells. Using the pharmacological reagent ethyl isopropyl amiloride (EIPA) to inhibit NHE1 activity and decrease pHi, we observe no change in glycolysis, as indicated by secreted lactate and intracellular pyruvate, despite confirming increased activity of the glycolytic enzyme phosphofructokinase-1 at higher pH. Also, in contrast to predictions, we find a significant decrease in oxidative phosphorylation with EIPA, as indicated by oxygen consumption rate (OCR). Decreased OCR with EIPA is not associated with changes in pathways that fuel oxidative phosphorylation or with mitochondrial membrane potential but occurs with a change in mitochondrial dynamics that includes a significant increase in elongated mitochondrial networks, suggesting increased fusion. These findings conflict with current paradigms on increased pHi inhibiting oxidative phosphorylation and increased oxidative phosphorylation being associated with mitochondrial fusion. Moreover, these findings raise questions on the suggested use of EIPA-like compounds to limit metabolic reprogramming in cancer cells.
    Keywords:  NHE1; cancer metabolism; glycolysis; intracellular pH; mitochondria
    DOI:  https://doi.org/10.1152/ajpcell.00001.2021
  2. Cell Rep. 2021 May 25. pii: S2211-1247(21)00525-8. [Epub ahead of print]35(8): 109180
    Mass-spectrometry-based proteomics reveals mitochondrial supercomplexome plasticity.
    Alba Gonzalez-Franquesa, Ben Stocks, Sabina Chubanava, Helle B Hattel, Roger Moreno-Justicia, Lone Peijs, Jonas T Treebak, Juleen R Zierath, Atul S Deshmukh.
      Mitochondrial respiratory complex subunits assemble in supercomplexes. Studies of supercomplexes have typically relied upon antibody-based quantification, often limited to a single subunit per respiratory complex. To provide a deeper insight into mitochondrial and supercomplex plasticity, we combine native electrophoresis and mass spectrometry to determine the supercomplexome of skeletal muscle from sedentary and exercise-trained mice. We quantify 422 mitochondrial proteins within 10 supercomplex bands in which we show the debated presence of complexes II and V. Exercise-induced mitochondrial biogenesis results in non-stoichiometric changes in subunits and incorporation into supercomplexes. We uncover the dynamics of supercomplex-related assembly proteins and mtDNA-encoded subunits after exercise. Furthermore, exercise affects the complexing of Lactb, an obesity-associated mitochondrial protein, and ubiquinone biosynthesis proteins. Knockdown of ubiquinone biosynthesis proteins leads to alterations in mitochondrial respiration. Our approach can be applied to broad biological systems. In this instance, comprehensively analyzing respiratory supercomplexes illuminates previously undetectable complexity in mitochondrial plasticity.
    Keywords:  complexome; exercise; mitochondrial respiratory complexes; mitochondrial supercomplexes; oxidative phosphorylation; protein complexes
    DOI:  https://doi.org/10.1016/j.celrep.2021.109180
  3. Sci Rep. 2021 May 25. 11(1): 10897
    Generation of somatic mitochondrial DNA-replaced cells for mitochondrial dysfunction treatment.
    Hideki Maeda, Daisuke Kami, Ryotaro Maeda, Akira Shikuma, Satoshi Gojo.
      Mitochondrial diseases currently have no cure regardless of whether the cause is a nuclear or mitochondrial genome mutation. Mitochondrial dysfunction notably affects a wide range of disorders in aged individuals, including neurodegenerative diseases, cancers, and even senescence. Here, we present a procedure to generate mitochondrial DNA-replaced somatic cells with a combination of a temporal reduction in endogenous mitochondrial DNA and coincubation with exogeneous isolated mitochondria. Heteroplasmy in mitochondrial disease patient-derived fibroblasts in which the mutant genotype was dominant over the wild-type genotype was reversed. Mitochondrial disease patient-derived fibroblasts regained respiratory function and showed lifespan extension. Mitochondrial membranous components were utilized as a vehicle to deliver the genetic materials into endogenous mitochondria-like horizontal genetic transfer in prokaryotes. Mitochondrial DNA-replaced cells could be a resource for transplantation to treat maternal inherited mitochondrial diseases.
    DOI:  https://doi.org/10.1038/s41598-021-90316-1
  4. J Biol Chem. 2021 May 19. pii: S0021-9258(21)00568-8. [Epub ahead of print] 100775
    Excess Exogenous Pyruvate Inhibits Lactate Dehydrogenase Activity in Live Cells in a MCT1-Dependent Manner.
    Yi Rao, Seth Gammon, Margie N Sutton, Niki M Zacharias, Pratip Bhattacharya, David Piwnica-Worms.
      Cellular pyruvate is an essential metabolite at the crossroads of glycolysis and oxidative phosphorylation, capable of supporting fermentative glycolysis by reduction to lactate mediated by lactate dehydrogenase (LDH) among other functions. Several inherited diseases of mitochondrial metabolism impact extracellular (plasma) pyruvate concentrations, and [1-13C]pyruvate infusion is used in isotope-labeled metabolic tracing studies, including hyperpolarized magnetic resonance spectroscopic imaging (MRSI). However, how these extracellular pyruvate sources impact intracellular metabolism is not clear. Herein, we examined the effects of excess exogenous pyruvate on intracellular LDH activity, extracellular acidification rates (ECAR) as a measure of lactate production, and hyperpolarized [1-13C]pyruvate-to-[1-13C]lactate conversion rates across a panel of tumor and normal cells. Combined LDH activity and LDHB/LDHA expression analysis intimated various hetero-tetrameric isoforms comprised of LDHA and LDHB in tumor cells, not only canonical LDHA. Millimolar concentrations of exogenous pyruvate induced substrate inhibition of LDH activity in both enzymatic assays ex vivo and in live cells, abrogated glycolytic ECAR, and inhibited hyperpolarized [1-13C]pyruvate-to-[1-13C]lactate conversion rates in cellulo. Importantly, the extent of exogenous pyruvate-induced inhibition of LDH and glycolytic ECAR in live cells was highly dependent on pyruvate influx, functionally mediated by monocarboxylate transporter-1 (MCT1) localized to the plasma membrane. These data provided evidence that highly concentrated bolus injections of pyruvate in vivo may transiently inhibit LDH activity in a tissue type- and MCT1-dependent manner. Maintaining plasma pyruvate at submillimolar concentrations could potentially minimize transient metabolic perturbations, improve pyruvate therapy, and enhance quantification of metabolic studies, including hyperpolarized [1-13C]pyruvate MRSI and stable isotope tracer experiments.
    Keywords:  (13)C magnetic resonance spectroscopy; LDH; MRSI; [1-(13)C]pyruvate; cancer metabolism; hyperpolarized (13)C; lactate dehydrogenase; stable isotope
    DOI:  https://doi.org/10.1016/j.jbc.2021.100775
  5. JCI Insight. 2021 May 25. pii: 142801. [Epub ahead of print]
    ECSIT is a critical limiting factor for cardiac function.
    Linan Xu, Fiachra Humphries, Nezira Delagic, Bingwei Wang, Ashling Holland, Kevin S Edgar, Jose R Hombrebueno, Donna Beer Stolz, Ewa Oleszycka, Aoife M Rodgers, Nadezhda Glezeva, Kenneth McDonald, Chris J Watson, Mark T Ledwidge, Rebecca J Ingram, David J Grieve, Paul N Moynagh.
      ECSIT is a protein with roles in early development, activation of the transcription factor NFB and production of mitochondrial reactive oxygen species (mROS) that facilitates clearance of intracellular bacteria like Salmonella. ECSIT is also an important assembly factor for mitochondrial complex I. Unlike the murine form of Ecsit (mEcsit), we demonstrate here that human ECSIT (hECSIT) to be highly labile. In order to explore if the instability of hECSIT affects functions previously ascribed to its murine counterpart, we created a novel transgenic mouse in which the murine Ecsit gene is replaced by the human ECSIT gene. The humanised mouse has low levels of hECSIT protein in keeping with its intrinsic instability. Whereas low level expression of hECSIT was capable of fully compensating for mEcsit in its roles in early development and activation of the NFB pathway, macrophages from humanised mice showed impaired clearance of Salmonella that was associated with reduced production of mROS. Notably, severe cardiac hypertrophy manifested in ageing humanised mice leading to premature death. The cellular and molecular basis to this phenotype is delineated by showing that low levels of human ECSIT protein leads to marked reduction in assembly and activity of mitochondrial complex I with impaired oxidative phosphorylation and reduced production of ATP. Cardiac tissue from humanised hECSIT mice also shows reduced mitochondrial fusion and more fission but impaired clearance of fragmented mitochondria. A cardiomyocyte-intrinsic role for Ecsit in mitochondrial function and cardioprotection is also demonstrated. We also show that cardiac fibrosis and damage in humans correlates with low expression of human ECSIT. In summary, our findings identify a new role for ECSIT in cardioprotection whilst also generating a valuable new experimental model to study mitochondrial dysfunction and cardiac pathophysiology.
    Keywords:  Cardiology; Cardiovascular disease; Metabolism; Mitochondria
    DOI:  https://doi.org/10.1172/jci.insight.142801
  6. J Physiol. 2021 May 25.
    Preserved skeletal muscle oxidative capacity in older adults despite decreased cardiorespiratory fitness with aging.
    Xiaoyan Zhang, Hawley E Kunz, Kevin Gries, Corey R Hart, Eric C Polley, Ian R Lanza.
      KEY POINTS: Healthy older adults exhibit lower cardiorespiratory fitness (VO2 peak) than young in the absence of any age-related difference in skeletal muscle mitochondrial capacity, suggesting central hemodynamics plays a larger role in age-related declines in VO2 peak. Total physical activity did not differ by age, but moderate-to-vigorous physical activity was lower in older compared to young adults. Moderate-to-vigorous physical activity is associated with VO2 peak and muscle oxidative capacity, but physical inactivity cannot entirely explain the age-related reduction in VO2 peak.ABSTRACT: Declining fitness (VO2 peak) is a hallmark of aging and believed to arise from decreased oxygen delivery and reduced muscle oxidative capacity. Physical activity is a modifiable lifestyle factor that is critical when evaluating the effects of age on parameters of fitness and energy metabolism. The objective was to evaluate the effects of age and sex on VO2 peak, muscle mitochondrial physiology, and physical activity in young and older adults. An additional objective was to assess the contribution of skeletal muscle oxidative capacity to age-related reductions in VO2 peak and determine if age-related variation in VO2 peak and muscle oxidative capacity could be explained on the basis of physical activity levels. 23 young and 52 older men and women completed measurements of VO2 peak, mitochondrial physiology in permeabilized muscle fibers, and free-living physical activity by accelerometry. Regression analyses were used to evaluate associations between age and VO2 peak, mitochondrial function, and physical activity. Significant age-related reductions were observed for VO2 peak (P<0.001), but not muscle mitochondrial capacity. Total daily step counts did not decrease with age, but older adults showed lower moderate-to-vigorous physical activity, which was associated with VO2 peak (R2 = 0.323, P<0.001) and muscle oxidative capacity (R2 = 0.086, P = 0.011). After adjusting for sex and physical activity, age was negatively associated with VO2 peak but not muscle oxidative capacity. Healthy older adults exhibit lower VO2 peak but preserved mitochondrial capacity compared to young. Physical activity, particularly moderate-to-vigorous, is a key factor in observed age-related changes in fitness and muscle oxidative capacity, but cannot entirely explain the age-related reduction in VO2 peak. This article is protected by copyright. All rights reserved.
    Keywords:  ageing; mitochondria; physical activity; skeletal muscle
    DOI:  https://doi.org/10.1113/JP281691
  7. Blood Adv. 2021 05 25. 5(10): 2490-2504
    Mitochondrial localization and moderated activity are key to murine erythroid enucleation.
    Raymond Liang, Vijay Menon, Jiajing Qiu, Tasleem Arif, Santosh Renuse, Miao Lin, Roberta Nowak, Boris Hartmann, Nikos Tzavaras, Deanna L Benson, Jerry E Chipuk, Miguel Fribourg, Akhilesh Pandey, Velia Fowler, Saghi Ghaffari.
      Mammalian red blood cells (RBCs), which primarily contain hemoglobin, exemplify an elaborate maturation process, with the terminal steps of RBC generation involving extensive cellular remodeling. This encompasses alterations of cellular content through distinct stages of erythroblast maturation that result in the expulsion of the nucleus (enucleation) followed by the loss of mitochondria and all other organelles and a transition to anaerobic glycolysis. Whether there is any link between erythroid removal of the nucleus and the function of any other organelle, including mitochondria, remains unknown. Here we demonstrate that mitochondria are key to nuclear clearance. Using live and confocal microscopy and high-throughput single-cell imaging, we show that before nuclear polarization, mitochondria progressively move toward one side of maturing erythroblasts and aggregate near the nucleus as it extrudes from the cell, a prerequisite for enucleation to proceed. Although we found active mitochondrial respiration is required for nuclear expulsion, levels of mitochondrial activity identify distinct functional subpopulations, because terminally maturing erythroblasts with low relative to high mitochondrial membrane potential are at a later stage of maturation, contain greatly condensed nuclei with reduced open chromatin-associated acetylation histone marks, and exhibit higher enucleation rates. Lastly, to our surprise, we found that late-stage erythroblasts sustain mitochondrial metabolism and subsequent enucleation, primarily through pyruvate but independent of in situ glycolysis. These findings demonstrate the critical but unanticipated functions of mitochondria during the erythroblast enucleation process. They are also relevant to the in vitro production of RBCs as well as to disorders of the erythroid lineage.
    DOI:  https://doi.org/10.1182/bloodadvances.2021004259
  8. Cancer Discov. 2021 May 26. pii: candisc.1437.2020. [Epub ahead of print]
    Medium-chain acyl CoA dehydrogenase protects mitochondria from lipid peroxidation in glioblastoma.
    Francesca Puca, Fei Yu, Caterina Bartolacci, Piergiorgio Pettazzoni, Alessandro Carugo, Emmet Huang-Hobbs, Jintan Liu, Ciro Zanca, Federica Carbone, Edoardo Del Poggetto, Joy Gumin, Pushan Dasgupta, Sahil Seth, Sanjana Srinivasan, Frederick F Lang, Erik P Sulman, Philip L Lorenzi, Lin Tan, Mengrou Shan, Zachary P Tolstyka, Maureen Kachman, Li Zhang, Sisi Gao, Angela K Deem, Giannicola Genovese, Pier Paolo Scaglioni, Costas A Lyssiotis, Andrea Viale, Giulio F Draetta.
      Glioblastoma (GBM) is highly resistant to chemo- and immune-based therapies and targeted inhibitors. To identify novel drug targets, we screened orthotopically implanted, patient-derived glioblastoma sphere-forming cells (GSCs) using an RNAi library to probe essential tumor cell metabolic programs. This identified high dependence on mitochondrial fatty acid metabolism. We focused on medium-chain acyl-CoA dehydrogenase (MCAD), which oxidizes medium-chain fatty acids (MCFAs), due to its consistently high score and high expression among models and upregulation in GBM compared to normal brain. Beyond the expected energetics impairment, MCAD depletion in primary GBM models induced an irreversible cascade of detrimental metabolic effects characterized by accumulation of unmetabolized MCFAs, which induced lipid peroxidation and oxidative stress, irreversible mitochondrial damage, and apoptosis. Our data uncover a novel protective role for MCAD to clear lipid molecules that may cause lethal cell damage, suggesting that therapeutic targeting of MCFA catabolism could exploit a key metabolic feature of GBM.
    DOI:  https://doi.org/10.1158/2159-8290.CD-20-1437
  9. Elife. 2021 May 26. pii: e67624. [Epub ahead of print]10
    Genome-wide CRISPRi screening identifies OCIAD1 as a prohibitin client and regulatory determinant of mitochondrial Complex III assembly in human cells.
    Maxence Le Vasseur, Jonathan Friedman, Marco Jost, Jiawei Xu, Justin Yamada, Martin Kampmann, Max A Horlbeck, Michelle R Salemi, Brett S Phinney, Jonathan S Weissman, Jodi Nunnari.
      Dysfunction of the mitochondrial electron transport chain (mETC) is a major cause of human mitochondrial diseases. To identify determinants of mETC function, we screened a genome-wide human CRISPRi library under oxidative metabolic conditions with selective inhibition of mitochondrial Complex III and identified ovarian carcinoma immunoreactive antigen (OCIA) domain-containing protein 1 (OCIAD1) as a Complex III assembly factor. We find that OCIAD1 is an inner mitochondrial membrane protein that forms a complex with supramolecular prohibitin assemblies. Our data indicate that OCIAD1 is required for maintenance of normal steady-state levels of Complex III and the proteolytic processing of the catalytic subunit cytochrome c1 (CYC1). In OCIAD1 depleted mitochondria, unprocessed CYC1 is hemylated and incorporated into Complex III. We propose that OCIAD1 acts as an adaptor within prohibitin assemblies to stabilize and/or chaperone CYC1 and to facilitate its proteolytic processing by the IMMP2L protease.
    Keywords:  Complex III; cell biology; cytochrome c1; electron transport chain; human; mitochondria; prohibitin; protease
    DOI:  https://doi.org/10.7554/eLife.67624
  10. Nat Commun. 2021 May 25. 12(1): 3108
    Sulfide catabolism ameliorates hypoxic brain injury.
    Eizo Marutani, Masanobu Morita, Shuichi Hirai, Shinichi Kai, Robert M H Grange, Yusuke Miyazaki, Fumiaki Nagashima, Lisa Traeger, Aurora Magliocca, Tomoaki Ida, Tetsuro Matsunaga, Daniel R Flicker, Benjamin Corman, Naohiro Mori, Yumiko Yamazaki, Annabelle Batten, Rebecca Li, Tomohiro Tanaka, Takamitsu Ikeda, Akito Nakagawa, Dmitriy N Atochin, Hideshi Ihara, Benjamin A Olenchock, Xinggui Shen, Motohiro Nishida, Kenjiro Hanaoka, Christopher G Kevil, Ming Xian, Donald B Bloch, Takaaki Akaike, Allyson G Hindle, Hozumi Motohashi, Fumito Ichinose.
      The mammalian brain is highly vulnerable to oxygen deprivation, yet the mechanism underlying the brain's sensitivity to hypoxia is incompletely understood. Hypoxia induces accumulation of hydrogen sulfide, a gas that inhibits mitochondrial respiration. Here, we show that, in mice, rats, and naturally hypoxia-tolerant ground squirrels, the sensitivity of the brain to hypoxia is inversely related to the levels of sulfide:quinone oxidoreductase (SQOR) and the capacity to catabolize sulfide. Silencing SQOR increased the sensitivity of the brain to hypoxia, whereas neuron-specific SQOR expression prevented hypoxia-induced sulfide accumulation, bioenergetic failure, and ischemic brain injury. Excluding SQOR from mitochondria increased sensitivity to hypoxia not only in the brain but also in heart and liver. Pharmacological scavenging of sulfide maintained mitochondrial respiration in hypoxic neurons and made mice resistant to hypoxia. These results illuminate the critical role of sulfide catabolism in energy homeostasis during hypoxia and identify a therapeutic target for ischemic brain injury.
    DOI:  https://doi.org/10.1038/s41467-021-23363-x
  11. Nat Rev Cancer. 2021 May 27.
    Mitochondrial DNA variation and cancer.
    Piotr K Kopinski, Larry N Singh, Shiping Zhang, Marie T Lott, Douglas C Wallace.
      Variation in the mitochondrial DNA (mtDNA) sequence is common in certain tumours. Two classes of cancer mtDNA variants can be identified: de novo mutations that act as 'inducers' of carcinogenesis and functional variants that act as 'adaptors', permitting cancer cells to thrive in different environments. These mtDNA variants have three origins: inherited variants, which run in families, somatic mutations arising within each cell or individual, and variants that are also associated with ancient mtDNA lineages (haplogroups) and are thought to permit adaptation to changing tissue or geographic environments. In addition to mtDNA sequence variation, mtDNA copy number and perhaps transfer of mtDNA sequences into the nucleus can contribute to certain cancers. Strong functional relevance of mtDNA variation has been demonstrated in oncocytoma and prostate cancer, while mtDNA variation has been reported in multiple other cancer types. Alterations in nuclear DNA-encoded mitochondrial genes have confirmed the importance of mitochondrial metabolism in cancer, affecting mitochondrial reactive oxygen species production, redox state and mitochondrial intermediates that act as substrates for chromatin-modifying enzymes. Hence, subtle changes in the mitochondrial genotype can have profound effects on the nucleus, as well as carcinogenesis and cancer progression.
    DOI:  https://doi.org/10.1038/s41568-021-00358-w
  12. Nat Metab. 2021 May;3(5): 665-681
    Mammary epithelial cells have lineage-rooted metabolic identities.
    Mathepan Jeya Mahendralingam, Hyeyeon Kim, Curtis William McCloskey, Kazeera Aliar, Alison Elisabeth Casey, Pirashaanthy Tharmapalan, Davide Pellacani, Vladimir Ignatchenko, Mar Garcia-Valero, Luis Palomero, Ankit Sinha, Jennifer Cruickshank, Ronak Shetty, Ravi N Vellanki, Marianne Koritzinsky, Vid Stambolic, Mina Alam, Aaron David Schimmer, Hal Kenneth Berman, Connie J Eaves, Miquel Angel Pujana, Thomas Kislinger, Rama Khokha.
      Cancer metabolism adapts the metabolic network of its tissue of origin. However, breast cancer is not a disease of a single origin. Multiple epithelial populations serve as the culprit cell of origin for specific breast cancer subtypes, yet our knowledge of the metabolic network of normal mammary epithelial cells is limited. Using a multi-omic approach, here we identify the diverse metabolic programmes operating in normal mammary populations. The proteomes of basal, luminal progenitor and mature luminal cell populations revealed enrichment of glycolysis in basal cells and of oxidative phosphorylation in luminal progenitors. Single-cell transcriptomes corroborated lineage-specific metabolic identities and additional intra-lineage heterogeneity. Mitochondrial form and function differed across lineages, with clonogenicity correlating with mitochondrial activity. Targeting oxidative phosphorylation and glycolysis with inhibitors exposed lineage-rooted metabolic vulnerabilities of mammary progenitors. Bioinformatics indicated breast cancer subtypes retain metabolic features of their putative cell of origin. Thus, lineage-rooted metabolic identities of normal mammary cells may underlie breast cancer metabolic heterogeneity and targeting these vulnerabilities could advance breast cancer therapy.
    DOI:  https://doi.org/10.1038/s42255-021-00388-6
  13. Sci Adv. 2021 May;pii: eabe7548. [Epub ahead of print]7(22):
    Elevated type I interferon responses potentiate metabolic dysfunction, inflammation, and accelerated aging in mtDNA mutator mice.
    Yuanjiu Lei, Camila Guerra Martinez, Sylvia Torres-Odio, Samantha L Bell, Christine E Birdwell, Joshua D Bryant, Carl W Tong, Robert O Watson, Laura Ciaccia West, A Phillip West.
      Mitochondrial dysfunction is a key driver of inflammatory responses in human disease. However, it remains unclear whether alterations in mitochondria-innate immune cross-talk contribute to the pathobiology of mitochondrial disorders and aging. Using the polymerase gamma (POLG) mutator model of mitochondrial DNA instability, we report that aberrant activation of the type I interferon (IFN-I) innate immune axis potentiates immunometabolic dysfunction, reduces health span, and accelerates aging in mutator mice. Mechanistically, elevated IFN-I signaling suppresses activation of nuclear factor erythroid 2-related factor 2 (NRF2), which increases oxidative stress, enhances proinflammatory cytokine responses, and accelerates metabolic dysfunction. Ablation of IFN-I signaling attenuates hyperinflammatory phenotypes by restoring NRF2 activity and reducing aerobic glycolysis, which combine to lessen cardiovascular and myeloid dysfunction in aged mutator mice. These findings further advance our knowledge of how mitochondrial dysfunction shapes innate immune responses and provide a framework for understanding mitochondria-driven immunopathology in POLG-related disorders and aging.
    DOI:  https://doi.org/10.1126/sciadv.abe7548
  14. STAR Protoc. 2021 Jun 18. 2(2): 100543
    Calibration and measurement of mitochondrial pH in intact adult rat cardiomyocytes.
    Meng Gao, Yuan Qin, Anqi Li, Hanyu Liu, Lei Chen, Bilin Liu, Ying Zhang, Yufei Gao, Guohua Gong.
      Mitochondrial pH is a vital parameter of the mitochondrial environment, which determines the rate of many mitochondrial functions, including metabolism, membrane potential, fate, etc. Abnormal mitochondrial pH is always closely related to the health status of cells. Analyzing mitochondrial pH can serve as a proxy for mitochondrial and cellular function. This protocol describes the use of SNARF-1 AM, a pH-sensitive fluorophore, to measure mitochondrial pH. This protocol details the steps to evaluate mitochondrial pH in live adult cardiomyocytes using confocal microscopy. The protocol can be adapted to other adherent cell types. For complete details on the use and execution of this protocol, please refer to Wei-LaPierre et al. (2013).
    Keywords:  Cell Biology; Metabolism; Microscopy; Molecular/Chemical Probes
    DOI:  https://doi.org/10.1016/j.xpro.2021.100543
  15. Arch Biochem Biophys. 2021 May 24. pii: S0003-9861(21)00183-1. [Epub ahead of print] 108934
    NADPH supply and the contribution of NAD(P)+ transhydrogenase (NNT) to H2O2 balance in skeletal muscle mitochondria.
    Tiago R Figueira, Annelise Francisco, Juliana A Ronchi, Guilherme R R M Dos Santos, William Dos Santos, Jason R Treberg, Roger F Castilho.
      H2O2 is endogenously generated and its removal in the matrix of skeletal muscle mitochondria (SMM) is dependent on NADPH likely provided by NAD(P)+ transhydrogenase (NNT) and isocitrate dehydrogenase (IDH2). Importantly, NNT activity is linked to mitochondrial protonmotive force. Here, we demonstrate the presence of NNT function in detergent-solubilized and intact functional SMM isolated from rats and wild type (Nnt+/+) mice, but not in SMM from congenic mice carrying a mutated NNT gene (Nnt-/-). Further comparisons between SMM from both Nnt mouse genotypes revealed that the NADPH supplied by NNT supports up to 600 pmol/mg/min of H2O2 removal under selected conditions. Surprisingly, SMM from Nnt-/- mice removed exogenous H2O2 at wild-type levels and exhibited a maintained or even decreased net emission of endogenous H2O2 when substrates that support Krebs cycle reactions were present (e.g., pyruvate plus malate or palmitoylcarnitine plus malate). These results may be explained by a compensation for the lack of NNT, since the total activities of concurrent NADP+-reducing enzymes (IDH2, malic enzymes and glutamate dehydrogenase) were ∼70% elevated in Nnt-/- mice. Importantly, respiratory rates were similar between SMM from both Nnt genotypes despite differing NNT contributions to H2O2 removal and their implications for an evolving concept in the literature are discussed. We concluded that NNT is capable of meaningfully sustaining NADPH-dependent H2O2 removal in intact SMM. Nonetheless, if the available substrates favor non-NNT sources of NADPH, the H2O2 removal by SMM is maintained in Nnt-/- mice SMM.
    Keywords:  Antioxidant; C57BL/6J; Krebs cycle; Oxidative stress; Redox balance
    DOI:  https://doi.org/10.1016/j.abb.2021.108934
  16. Nat Commun. 2021 May 27. 12(1): 3188
    Chromosomal copy number heterogeneity predicts survival rates across cancers.
    Erik van Dijk, Tom van den Bosch, Kristiaan J Lenos, Khalid El Makrini, Lisanne E Nijman, Hendrik F B van Essen, Nico Lansu, Michiel Boekhout, Joris H Hageman, Rebecca C Fitzgerald, Cornelis J A Punt, Jurriaan B Tuynman, Hugo J G Snippert, Geert J P L Kops, Jan Paul Medema, Bauke Ylstra, Louis Vermeulen, Daniël M Miedema.
      Survival rates of cancer patients vary widely within and between malignancies. While genetic aberrations are at the root of all cancers, individual genomic features cannot explain these distinct disease outcomes. In contrast, intra-tumour heterogeneity (ITH) has the potential to elucidate pan-cancer survival rates and the biology that drives cancer prognosis. Unfortunately, a comprehensive and effective framework to measure ITH across cancers is missing. Here, we introduce a scalable measure of chromosomal copy number heterogeneity (CNH) that predicts patient survival across cancers. We show that the level of ITH can be derived from a single-sample copy number profile. Using gene-expression data and live cell imaging we demonstrate that ongoing chromosomal instability underlies the observed heterogeneity. Analysing 11,534 primary cancer samples from 37 different malignancies, we find that copy number heterogeneity can be accurately deduced and predicts cancer survival across tissues of origin and stages of disease. Our results provide a unifying molecular explanation for the different survival rates observed between cancer types.
    DOI:  https://doi.org/10.1038/s41467-021-23384-6
  17. Blood Cancer Discov. 2021 May;2(3): 266-287
    SIRT5 IS A DRUGGABLE METABOLIC VULNERABILITY IN ACUTE MYELOID LEUKEMIA.
    Dongqing Yan, Anca Franzini, Anthony D Pomicter, Brayden J Halverson, Orlando Antelope, Clinton C Mason, Jonathan M Ahmann, Anna V Senina, Nadeem A Vellore, Courtney L Jones, Matthew S Zabriskie, Hein Than, Michael J Xiao, Alexandria van Scoyk, Ami B Patel, Phillip M Clair, William L Heaton, Shawn C Owen, Joshua L Andersen, Christina M Egbert, Julie A Reisz, Angelo D'Alessandro, James E Cox, Kevin C Gantz, Hannah M Redwine, Siddharth M Iyer, Jamshid S Khorashad, Nima Rajabi, Christian A Olsen, Thomas O'Hare, Michael W Deininger.
      We discovered that the survival and growth of many primary acute myeloid leukemia (AML) samples and cell lines, but not normal CD34+ cells, are dependent on SIRT5, a lysine deacylase implicated in regulating multiple metabolic pathways. Dependence on SIRT5 is genotype-agnostic and extends to RAS- and p53-mutated AML. Results were comparable between SIRT5 knockdown and SIRT5 inhibition using NRD167, a potent and selective SIRT5 inhibitor. Apoptosis induced by SIRT5 disruption is preceded by reductions in oxidative phosphorylation and glutamine utilization, and an increase in mitochondrial superoxide that is attenuated by ectopic superoxide dismutase 2. These data indicate that SIRT5 controls and coordinates several key metabolic pathways in AML and implicate SIRT5 as a vulnerability in AML.
    DOI:  https://doi.org/10.1158/2643-3230.bcd-20-0168
  18. Nat Immunol. 2021 Jun;22(6): 746-756
    Metabolic reprogramming of terminally exhausted CD8+ T cells by IL-10 enhances anti-tumor immunity.
    Yugang Guo, Yu-Qing Xie, Min Gao, Yang Zhao, Fabien Franco, Mathias Wenes, Imran Siddiqui, Alessio Bevilacqua, Haiping Wang, Hanshuo Yang, Bing Feng, Xin Xie, Catherine M Sabatel, Benjamin Tschumi, Amphun Chaiboonchoe, Yuxi Wang, Weimin Li, Weihua Xiao, Werner Held, Pedro Romero, Ping-Chih Ho, Li Tang.
      T cell exhaustion presents one of the major hurdles to cancer immunotherapy. Among exhausted CD8+ tumor-infiltrating lymphocytes, the terminally exhausted subset contributes directly to tumor cell killing owing to its cytotoxic effector function. However, this subset does not respond to immune checkpoint blockades and is difficult to be reinvigorated with restored proliferative capacity. Here, we show that a half-life-extended interleukin-10-Fc fusion protein directly and potently enhanced expansion and effector function of terminally exhausted CD8+ tumor-infiltrating lymphocytes by promoting oxidative phosphorylation, a process that was independent of the progenitor exhausted T cells. Interleukin-10-Fc was a safe and highly efficient metabolic intervention that synergized with adoptive T cell transfer immunotherapy, leading to eradication of established solid tumors and durable cures in the majority of treated mice. These findings show that metabolic reprogramming by upregulating mitochondrial pyruvate carrier-dependent oxidative phosphorylation can revitalize terminally exhausted T cells and enhance the response to cancer immunotherapy.
    DOI:  https://doi.org/10.1038/s41590-021-00940-2
  19. Cancer Gene Ther. 2021 May 27.
    Fatty acid β-oxidation promotes breast cancer stemness and metastasis via the miRNA-328-3p-CPT1A pathway.
    Feng Zeng, Mingkang Yao, Yun Wang, Wei Zheng, Shengshan Liu, Zeyu Hou, Xiaoming Cheng, Suhong Sun, Taolang Li, Hongyuan Zhao, Yi Luo, Jiang Li.
      MicroRNAs (miRNA) have been shown to be associated with tumor diagnosis, prognosis, and therapeutic response. MiR-328-3p plays a significant role in breast cancer growth; however, its actual function and how it modulates specific biological functions is poorly understood. Here, miR-328-3p was significantly downregulated in breast cancer, especially in patients with metastasis. Mitochondrial carnitine palmitoyl transferase 1a (CPT1A) is a downstream target gene in the miR-328-3p-regulated pathway. Furthermore, the miR-328-3p/CPT1A/fatty acid β-oxidation/stemness axis was shown responsible for breast cancer metastasis. Collectively, this study revealed that miR-328-3p is a potential therapeutic target for the treatment of breast cancer patients with metastasis, and also a model for the miRNA-fatty acid β-oxidation-stemness axis, which may assist inunderstanding the cancer stem cell signaling functions of miRNA.
    DOI:  https://doi.org/10.1038/s41417-021-00348-y
  20. Sci Rep. 2021 May 24. 11(1): 10753
    Iron supplementation regulates the progression of high fat diet induced obesity and hepatic steatosis via mitochondrial signaling pathways.
    Naho Kitamura, Yoko Yokoyama, Hiroki Taoka, Utana Nagano, Shotaro Hosoda, Tanon Taworntawat, Anna Nakamura, Yoko Ogawa, Kazuo Tsubota, Mitsuhiro Watanabe.
      Disruption of iron metabolism is closely related to metabolic diseases. Iron deficiency is frequently associated with obesity and hepatic steatosis. However, the effects of iron supplementation on obesity and energy metabolism remain unclear. Here we show that a high-fat diet supplemented with iron reduces body weight gain and hepatic lipid accumulation in mice. Iron supplementation was found to reduce mitochondrial morphological abnormalities and upregulate gene transcription involved in mitochondrial function and beta oxidation in the liver and skeletal muscle. In both these tissues, iron supplementation increased the expression of genes involved in heme or iron-sulfur (Fe-S) cluster synthesis. Heme and Fe-S cluster, which are iron prosthetic groups contained in electron transport chain complex subunits, are essential for mitochondrial respiration. The findings of this study demonstrated that iron regulates mitochondrial signaling pathways-gene transcription of mitochondrial component molecules synthesis and their energy metabolism. Overall, the study elucidates the molecular basis underlying the relationship between iron supplementation and obesity and hepatic steatosis progression, and the role of iron as a signaling molecule.
    DOI:  https://doi.org/10.1038/s41598-021-89673-8
  21. Front Oncol. 2021 ;11 672781
    The Functions, Methods, and Mobility of Mitochondrial Transfer Between Cells.
    Yiming Qin, Xin Jiang, Qi Yang, Jiaqi Zhao, Qiong Zhou, Yanhong Zhou.
      Mitochondria are vital organelles in cells, regulating energy metabolism and apoptosis. Mitochondrial transcellular transfer plays a crucial role during physiological and pathological conditions, such as rescuing recipient cells from bioenergetic deficit and tumorigenesis. Studies have shown several structures that conduct transcellular transfer of mitochondria, including tunneling nanotubes (TNTs), extracellular vesicles (EVs), and Cx43 gap junctions (GJs). The intra- and intercellular transfer of mitochondria is driven by a transport complex. Mitochondrial Rho small GTPase (MIRO) may be the adaptor that connects the transport complex with mitochondria, and myosin XIX is the motor protein of the transport complex, which participates in the transcellular transport of mitochondria through TNTs. In this review, the roles of TNTs, EVs, GJs, and related transport complexes in mitochondrial transcellular transfer are discussed in detail, as well as the formation mechanisms of TNTs and EVs. This review provides the basis for the development of potential clinical therapies targeting the structures of mitochondrial transcellular transfer.
    Keywords:  Cx43 gap junction; Miro; extracellular vesicles; mitochondria; myosin XIX; transcellular transport; tunneling nanotubes
    DOI:  https://doi.org/10.3389/fonc.2021.672781
  22. J Transl Med. 2021 May 24. 19(1): 219
    Pan-cancer characterization of metabolism-related biomarkers identifies potential therapeutic targets.
    Guoshu Bi, Yunyi Bian, Jiaqi Liang, Jiacheng Yin, Runmei Li, Mengnan Zhao, Yiwei Huang, Tao Lu, Cheng Zhan, Hong Fan, Qun Wang.
      BACKGROUND: Generally, cancer cells undergo metabolic reprogramming to adapt to energetic and biosynthetic requirements that support their uncontrolled proliferation. However, the mutual relationship between two critical metabolic pathways, glycolysis and oxidative phosphorylation (OXPHOS), remains poorly defined.METHODS: We developed a "double-score" system to quantify glycolysis and OXPHOS in 9668 patients across 33 tumor types from The Cancer Genome Atlas and classified them into four metabolic subtypes. Multi-omics bioinformatical analyses was conducted to detect metabolism-related molecular features.
    RESULTS: Compared with patients with low glycolysis and high OXPHOS (LGHO), those with high glycolysis and low OXPHOS (HGLO) were consistently associated with worse prognosis. We identified common dysregulated molecular features between different metabolic subgroups across multiple cancers, including gene, miRNA, transcription factor, methylation, and somatic alteration, as well as investigated their mutual interfering relationships.
    CONCLUSION: Overall, this work provides a comprehensive atlas of metabolic heterogeneity on a pan-cancer scale and identified several potential drivers of metabolic rewiring, suggesting corresponding prognostic and therapeutic utility.
    Keywords:  Glycolysis; Metabolism; Oxidative phosphorylation; Pan-cancer; Warburg effect
    DOI:  https://doi.org/10.1186/s12967-021-02889-0
  23. Diabetes. 2021 May 26. pii: db210037. [Epub ahead of print]
    Mitochondrial efflux of citrate and isocitrate is fully dispensable for glucose-stimulated insulin secretion and pancreatic islet β-cell function.
    Casey J Bauchle, Kristen E Rohli, Cierra K Boyer, Vidhant Pal, Jonathan V Rocheleau, Siming Liu, Yumi Imai, Eric B Taylor, Samuel B Stephens.
      The defining feature of pancreatic islet β-cell function is the precise coordination of changes in blood glucose levels with insulin secretion to regulate systemic glucose homeostasis. While ATP has long been heralded as a critical metabolic coupling factor to trigger insulin release, glucose-derived metabolites have been suggested to further amplify fuel-stimulated insulin secretion. The mitochondrial export of citrate and isocitrate through the citrate-isocitrate carrier (CIC) has been suggested to initiate a key pathway that amplifies glucose-stimulated insulin secretion, though the physiological significance of β-cell CIC to glucose homeostasis has not been established. Here, we generated constitutive and adult CIC β-cell knockout mice and demonstrate these animals have normal glucose tolerance, similar responses to diet-induced obesity, and identical insulin secretion responses to various fuel secretagogues. Glucose-stimulated NADPH production was impaired in β-cell CIC KO islets, whereas glutathione reduction was retained. Furthermore, suppression of the downstream enzyme, cytosolic isocitrate dehydrogenase, Idh1, inhibited insulin secretion in wild type islets, but failed to impact β-cell function in β-cell CIC KO islets. Our data demonstrate that the mitochondrial citrate-isocitrate carrier is not required for glucose-stimulated insulin secretion, and that additional complexities exist for the role of Idh1 and NADPH in the regulation of β-cell function.
    DOI:  https://doi.org/10.2337/db21-0037
  24. Nat Commun. 2021 May 28. 12(1): 3239
    Structures of the human LONP1 protease reveal regulatory steps involved in protease activation.
    Mia Shin, Edmond R Watson, Albert S Song, Jeffrey T Mindrebo, Scott J Novick, Patrick R Griffin, R Luke Wiseman, Gabriel C Lander.
      The human mitochondrial AAA+ protein LONP1 is a critical quality control protease involved in regulating diverse aspects of mitochondrial biology including proteostasis, electron transport chain activity, and mitochondrial transcription. As such, genetic or aging-associated imbalances in LONP1 activity are implicated in pathologic mitochondrial dysfunction associated with numerous human diseases. Despite this importance, the molecular basis for LONP1-dependent proteolytic activity remains poorly defined. Here, we solved cryo-electron microscopy structures of human LONP1 to reveal the underlying molecular mechanisms governing substrate proteolysis. We show that, like bacterial Lon, human LONP1 adopts both an open and closed spiral staircase orientation dictated by the presence of substrate and nucleotide. Unlike bacterial Lon, human LONP1 contains a second spiral staircase within its ATPase domain that engages substrate as it is translocated toward the proteolytic chamber. Intriguingly, and in contrast to its bacterial ortholog, substrate binding within the central ATPase channel of LONP1 alone is insufficient to induce the activated conformation of the protease domains. To successfully induce the active protease conformation in substrate-bound LONP1, substrate binding within the protease active site is necessary, which we demonstrate by adding bortezomib, a peptidomimetic active site inhibitor of LONP1. These results suggest LONP1 can decouple ATPase and protease activities depending on whether AAA+ or both AAA+ and protease domains bind substrate. Importantly, our structures provide a molecular framework to define the critical importance of LONP1 in regulating mitochondrial proteostasis in health and disease.
    DOI:  https://doi.org/10.1038/s41467-021-23495-0
  25. iScience. 2021 May 21. 24(5): 102434
    Chronic cold exposure induces autophagy to promote fatty acid oxidation, mitochondrial turnover, and thermogenesis in brown adipose tissue.
    Winifred W Yau, Kiraely Adam Wong, Jin Zhou, Nivetha Kanakaram Thimmukonda, Yajun Wu, Boon-Huat Bay, Brijesh Kumar Singh, Paul Michael Yen.
      Autophagy plays an important role in lipid breakdown, mitochondrial turnover, and mitochondrial function during brown adipose tissue (BAT) activation by thyroid hormone, but its role in BAT during adaptive thermogenesis remains controversial. Here, we examined BAT from mice exposed to 72 h of cold challenge as well as primary brown adipocytes treated with norepinephrine and found increased autophagy as well as increased β-oxidation, mitophagy, mitochondrial turnover, and mitochondrial activity. To further understand the role of autophagy of BAT in vivo, we generated BAT-specific Atg5 knockout (Atg5cKO) mice and exposed them to cold for 72 h. Interestingly, BAT-specific Atg5cKO mice were unable to maintain body temperature after chronic cold exposure and displayed deranged mitochondrial morphology and reactive oxygen species damage in their BAT. Our findings demonstrate the critical role of autophagy in adaptive thermogenesis, fatty acid metabolism, and mitochondrial function in BAT during chronic cold exposure.
    Keywords:  Cell Biology; Endocrine System Physiology; Molecular Physiology
    DOI:  https://doi.org/10.1016/j.isci.2021.102434
  26. Oncol Rep. 2021 Jul;pii: 136. [Epub ahead of print]46(1):
    ND‑09 inhibits chronic myeloid leukemia K562 cell growth by regulating BCR‑ABL signaling.
    Yan-Hong Liu, Man Zhu, Pan-Pan Lei, Xiao-Yan Pan, Wei-Na Ma.
      Chronic myeloid leukemia (CML) accounts for approximately 15% of new adult leukemia cases. The fusion gene BCR‑ABL is an important biological basis and target for CML. In the present study, a novel compound, ND‑09, was developed and its inhibitory effect and mechanism of action on CML growth were evaluated using RT‑PCR and western blot analysis. The results showed that ND‑09 demonstrated a high level of inhibitory action toward CML cells overexpressing BCR‑ABL and induced K562 cell apoptosis through the mitochondrial pathway. Notably, combined ND‑09 and BCR‑ABL siRNA treatment could better inhibit cell proliferation and induce apoptosis in K562 cells. Furthermore, this growth effect of BCR‑ABL siRNA could be fully rescued by transfection with BCR‑ABL. ND‑09 exhibited a good fit within BCR‑ABL and occupied its ATP‑binding pocket, thus altering BCR‑ABL kinase activity. Therefore, ND‑09 downregulated the phosphorylation of BCR‑ABL and ABL, ultimately inhibiting the downstream signaling pathways in K562 cells. These findings suggest that ND‑09 induces growth arrest in CML cells by targeting BCR‑ABL.
    Keywords:  BCR‑ABL; ND‑09; cell apoptosis; cell cycle; chronic myeloid leukemia
    DOI:  https://doi.org/10.3892/or.2021.8087
  27. Biochem Biophys Res Commun. 2021 May 22. pii: S0006-291X(21)00820-2. [Epub ahead of print]562 55-61
    Targeting NF-κB-dependent alkaliptosis for the treatment of venetoclax-resistant acute myeloid leukemia cells.
    Shan Zhu, Jiao Liu, Rui Kang, Minghua Yang, Daolin Tang.
      Venetoclax is a highly selective BCL2 inhibitor widely used in the treatment of leukemia, especially chronic lymphocytic leukemia and acute myeloid leukemia (AML). However, long-term use of venetoclax may lead to secondary drug resistance, which constitutes an important obstacle to prolonging the duration of the therapeutic response. Here, we show that the acquired resistance to venetoclax in human AML cell lines depends on NF-κB activation rather than on the upregulation of anti-apoptotic BCL2L1 expression. Moreover, alkaliptosis induced by the small molecular compound JTC801, but not necroptosis and ferroptosis, inhibits the growth of venetoclax-resistant AML cells in vitro and in xenograft mouse models. Mechanistically, NF-κB-mediated CA9 downregulation is required for intracellular pH upregulation, thereby inducing alkaliptosis in venetoclax-resistant cells. These findings provide a new strategy to selectively remove venetoclax-resistant AML cells.
    Keywords:  Acute myeloid leukemia; Alkaliptosis; BCL2; Drug resistance; Venetoclax
    DOI:  https://doi.org/10.1016/j.bbrc.2021.05.049
  28. J Nanobiotechnology. 2021 May 22. 19(1): 152
    pH-activated, mitochondria-targeted, and redox-responsive delivery of paclitaxel nanomicelles to overcome drug resistance and suppress metastasis in lung cancer.
    He Wang, Wenwen Shi, Danning Zeng, Qiudi Huang, Jiacui Xie, Huaying Wen, Jinfang Li, Xiyong Yu, Linghao Qin, Yi Zhou.
      BACKGROUND: Mitochondria play a role in the occurrence, development, drug resistance, metastasis, and other functions of cancer and thus are a drug target. An acid-activated mitochondria-targeting drug nanocarrier with redox-responsive function was constructed in the present study. However, whether this vector can precisely delivery paclitaxel (PTX) to enhance therapeutic efficacy in drug-resistant lung cancer is unknown.RESULTS: Acid-cleavable dimethylmaleic anhydride (DA) was used to modify pluronic P85-conjugated mitochondria-targeting triphenylphosphonium (TPP) using disulfide bonds as intermediate linkers (DA-P85-SS-TPP and DA-P-SS-T). The constructed nanocarriers demonstrated enhanced cellular uptake and selective mitochondrial targeting at extracellular pH characteristic for a tumor (6.5) and were characterized by extended circulation in the blood. TPP promoted the targeting of the DA-P-SS-T/PTX nanomicelles to the mitochondrial outer membrane to decrease the membrane potential and ATP level, resulting in inhibition of P-glycoprotein and suppression of drug resistance and cancer metastasis. PTX was also rapidly released in the presence of high glutathione (GSH) levels and directly diffused into the mitochondria, resulting in apoptosis of drug-resistant lung cancer cells.
    CONCLUSIONS: These promising results indicated that acid-activated mitochondria-targeting and redox-responsive nanomicelles potentially represent a significant advancement in cancer treatment. GRAPHIC ABSTARCT.
    Keywords:  Drug resistance; Lung cancer; Metastasis; Redox-responsive; pH-Activated Mitochondria-Targeted Delivery
    DOI:  https://doi.org/10.1186/s12951-021-00895-4
  29. Life Sci. 2021 May 19. pii: S0024-3205(21)00600-7. [Epub ahead of print]278 119614
    High doses of butyrate induce a reversible body temperature drop through transient proton leak in mitochondria of brain neurons.
    Yanhong Xu, Shiqiao Peng, Xinyu Cao, Shengnan Qian, Shuang Shen, Juntao Luo, Xiaoying Zhang, Hongbin Sun, Wei L Shen, Weiping Jia, Jianping Ye.
      AIMS: Sodium butyrate (SB) is a major product of gut microbiota with signaling activity in the human body. It has become a dietary supplement in the treatment of intestinal disorders. However, the toxic effect of overdosed SB and treatment strategy remain unknown. The two issues are addressed in current study.MATERIALS AND METHODS: SB (0.3-2.5 g/kg) was administrated through a single peritoneal injection in mice. The core body temperature and mitochondrial function in the brown adipose tissue and brain were monitored. Pharmacodynamics, targeted metabolomics, electron microscope, oxygen consumption rate and gene knockdown were employed to dissect the mechanism for the toxic effect.
    KEY FINDINGS: The temperature was reduced by SB (1.2-2.5 g/kg) in a dose-dependent manner in mice for 2-4 h. In the brain, the effect was associated with SB elevation and neurotransmitter reduction. Metabolites changes were seen in the glycolysis, TCA cycle and pentose phosphate pathways. Adenine nucleotide translocase (ANT) was activated by butyrate for proton transportation leading to a transient potential collapse through proton leak. The SB activity was attenuated by ANT inhibition from gene knockdown or pharmacological blocker. ROS was elevated by SB for the increased ANT activity in proton leak in Neuro-2a.
    SIGNIFICANCE: Excessive SB generated an immediate and reversible toxic effect for inhibition of body temperature through transient mitochondrial dysfunction in the brain. The mechanism was quick activation of ANT proteins for potential collapse in mitochondria. ROS may be a factor in the ANT activation by SB.
    Keywords:  ANT; Energy metabolism; MPTP; Mitochondria; Sodium butyrate
    DOI:  https://doi.org/10.1016/j.lfs.2021.119614
  30. Sci Rep. 2021 May 25. 11(1): 10925
    Single channel properties of mitochondrial large conductance potassium channel formed by BK-VEDEC splice variant.
    Shur Gałecka, Bogusz Kulawiak, Piotr Bednarczyk, Harpreet Singh, Adam Szewczyk.
      The activation of mitochondrial large conductance calcium-activated potassium (mitoBKCa) channels increases cell survival during ischemia/reperfusion injury of cardiac cells. The basic biophysical and pharmacological properties of mitoBKCa correspond to the properties of the BKCa channels from the plasma membrane. It has been suggested that the VEDEC splice variant of the KCNMA1 gene product encoding plasma membrane BKCa is targeted toward mitochondria. However there has been no direct evidence that this protein forms a functional channel in mitochondria. In our study, we used HEK293T cells to express the VEDEC splice variant and observed channel activity in mitochondria using the mitoplast patch-clamp technique. For the first time, we found that transient expression with the VEDEC isoform resulted in channel activity with the conductance of 290 ± 3 pS. The channel was voltage-dependent and activated by calcium ions. Moreover, the activity of the channel was stimulated by the potassium channel opener NS11021 and inhibited by hemin and paxilline, which are known BKCa channel blockers. Immunofluorescence experiments confirmed the partial colocalization of the channel within the mitochondria. From these results, we conclude that the VEDEC isoform of the BKCa channel forms a functional channel in the inner mitochondrial membrane. Additionally, our data show that HEK293T cells are a promising experimental model for expression and electrophysiological studies of mitochondrial potassium channels.
    DOI:  https://doi.org/10.1038/s41598-021-90465-3
  31. Cancer Cell. 2021 May 12. pii: S1535-6108(21)00225-7. [Epub ahead of print]
    N6-Methyladenosine on mRNA facilitates a phase-separated nuclear body that suppresses myeloid leukemic differentiation.
    Yuanming Cheng, Wei Xie, Brian F Pickering, Karen L Chu, Angela M Savino, Xuejing Yang, Hanzhi Luo, Diu Tt Nguyen, Shanlan Mo, Ersilia Barin, Anthony Velleca, Thomas M Rohwetter, Dinshaw J Patel, Samie R Jaffrey, Michael G Kharas.
      N6-Methyladenosine (m6A) on mRNAs mediates different biological processes and its dysregulation contributes to tumorigenesis. How m6A dictates its diverse molecular and cellular effects in leukemias remains unknown. We found that YTHDC1 is the essential m6A reader in myeloid leukemia from a genome-wide CRISPR screen and that m6A is required for YTHDC1 to undergo liquid-liquid phase separation and form nuclear YTHDC1-m6A condensates (nYACs). The number of nYACs increases in acute myeloid leukemia (AML) cells compared with normal hematopoietic stem and progenitor cells. AML cells require the nYACs to maintain cell survival and the undifferentiated state that is critical for leukemia maintenance. Furthermore, nYACs enable YTHDC1 to protect m6A-mRNAs from the PAXT complex and exosome-associated RNA degradation. Collectively, m6A is required for the formation of a nuclear body mediated by phase separation that maintains mRNA stability and control cancer cell survival and differentiation.
    Keywords:  RNA methylation; RNA-binding proteins; differentiation; myeloid leukemia; phase separation
    DOI:  https://doi.org/10.1016/j.ccell.2021.04.017
  32. Nat Commun. 2021 May 26. 12(1): 3175
    CaMKII oxidation is a critical performance/disease trade-off acquired at the dawn of vertebrate evolution.
    Qinchuan Wang, Erick O Hernández-Ochoa, Meera C Viswanathan, Ian D Blum, Danh C Do, Jonathan M Granger, Kevin R Murphy, An-Chi Wei, Susan Aja, Naili Liu, Corina M Antonescu, Liliana D Florea, C Conover Talbot, David Mohr, Kathryn R Wagner, Sergi Regot, Richard M Lovering, Peisong Gao, Mario A Bianchet, Mark N Wu, Anthony Cammarato, Martin F Schneider, Gabriel S Bever, Mark E Anderson.
      Antagonistic pleiotropy is a foundational theory that predicts aging-related diseases are the result of evolved genetic traits conferring advantages early in life. Here we examine CaMKII, a pluripotent signaling molecule that contributes to common aging-related diseases, and find that its activation by reactive oxygen species (ROS) was acquired more than half-a-billion years ago along the vertebrate stem lineage. Functional experiments using genetically engineered mice and flies reveal ancestral vertebrates were poised to benefit from the union of ROS and CaMKII, which conferred physiological advantage by allowing ROS to increase intracellular Ca2+ and activate transcriptional programs important for exercise and immunity. Enhanced sensitivity to the adverse effects of ROS in diseases and aging is thus a trade-off for positive traits that facilitated the early and continued evolutionary success of vertebrates.
    DOI:  https://doi.org/10.1038/s41467-021-23549-3
  33. Bioeng Transl Med. 2021 May;6(2): e10209
    Mitochondrial transplantation therapy inhibit carbon tetrachloride-induced liver injury through scavenging free radicals and protecting hepatocytes.
    Zizhen Zhao, Yixue Hou, Wei Zhou, Rajendiran Keerthiga, Ailing Fu.
      Carbon tetrachloride (CCl4)-induced liver injury is predominantly caused by free radicals, in which mitochondrial function of hepatocytes is impaired, accompanying with the production of ROS and decreased ATP energy supply in animals intoxicated with CCl4. Here we explored a novel therapeutic approach, mitochondrial transplantation therapy, for treating the liver injury. The results showed that mitochondria entered hepatocytes through macropinocytosis pathway, and thereby cell viability was recovered in a concentration-dependent manner. Mitochondrial therapy could increase ATP supply and reduce free radical damage. In liver injury model of mice, mitochondrial therapy significantly improved liver function and prevented tissue fibrogenesis. Transcriptomic data revealed that mitochondrial unfold protein response (UPRmt), a protective transcriptional response of mitochondria-to-nuclear retrograde signaling, would be triggered after mitochondrial administration. Then the anti-oxidant genes were up-regulated to scavenge free radicals. The mitochondrial function was rehabilitated through the transcriptional activation of respiratory chain enzyme and mitophage-associated genes. The protective response re-balanced the cellular homeostasis, and eventually enhanced stress resistance that is linked to cell survival. The efficacy of mitochondrial transplantation therapy in the animals would suggest a novel approach for treating liver injury caused by toxins.
    Keywords:  UPRmt; energy supply; free radical; mitochondrial therapy
    DOI:  https://doi.org/10.1002/btm2.10209
  34. FASEB J. 2021 Jun;35(6): e21680
    Extracellular vesicles activate ATM-Chk2 signaling pathway through the intercellular transfer of mitochondrial DNA in HBV-infected human hepatocytes.
    Hiromichi Dansako, Youki Ueda, Shinya Satoh, Nobuyuki Kato.
      Hepatitis B virus (HBV) is a human hepatotropic pathogen causing hepatocellular carcinoma. We recently obtained HBV-susceptible immortalized human hepatocyte NKNT-3 by exogenously expressing NTCP and its derived cell clones, #28.3.8 and #28.3.25.13 exhibiting different levels of HBV susceptibility. In the present study, we showed that HBV infection activated the ATM-Chk2 signaling pathway in #28.3.25.13 cells but not in #28.3.8 cells. Both the cell culture supernatant and extracellular vesicles (EVs) derived from HBV-infected #28.3.25.13 cells also activated the ATM-Chk2 signaling pathway in naïve #28.3.25.13 cells. Interestingly, EVs derived from HBV-infected #28.3.25.13 cells included higher level of mitochondrial DNA (mtDNA) than those from HBV-infected #28.3.8 cells. Based on our results, we propose the novel model that EVs mediate the activation of ATM-Chk2 signaling pathway by the intercellular transfer of mtDNA in HBV-infected human hepatocyte.
    Keywords:  ATM-Chk2 signaling pathway; extracellular vesicles; hepatitis B virus; mitochondrial DNA
    DOI:  https://doi.org/10.1096/fj.202002678R
  35. Nat Metab. 2021 May;3(5): 588-589
    Mitochondrial ROS signals prevent excessive immune response.
    Kim Zarse, Michael Ristow.
      
    DOI:  https://doi.org/10.1038/s42255-021-00387-7
  36. Nat Commun. 2021 May 28. 12(1): 3208
    Restoration of energy homeostasis by SIRT6 extends healthy lifespan.
    A Roichman, S Elhanati, M A Aon, I Abramovich, A Di Francesco, Y Shahar, M Y Avivi, M Shurgi, A Rubinstein, Y Wiesner, A Shuchami, Z Petrover, I Lebenthal-Loinger, O Yaron, A Lyashkov, C Ubaida-Mohien, Y Kanfi, B Lerrer, P J Fernández-Marcos, M Serrano, E Gottlieb, R de Cabo, H Y Cohen.
      Aging leads to a gradual decline in physical activity and disrupted energy homeostasis. The NAD+-dependent SIRT6 deacylase regulates aging and metabolism through mechanisms that largely remain unknown. Here, we show that SIRT6 overexpression leads to a reduction in frailty and lifespan extension in both male and female B6 mice. A combination of physiological assays, in vivo multi-omics analyses and 13C lactate tracing identified an age-dependent decline in glucose homeostasis and hepatic glucose output in wild type mice. In contrast, aged SIRT6-transgenic mice preserve hepatic glucose output and glucose homeostasis through an improvement in the utilization of two major gluconeogenic precursors, lactate and glycerol. To mediate these changes, mechanistically, SIRT6 increases hepatic gluconeogenic gene expression, de novo NAD+ synthesis, and systemically enhances glycerol release from adipose tissue. These findings show that SIRT6 optimizes energy homeostasis in old age to delay frailty and preserve healthy aging.
    DOI:  https://doi.org/10.1038/s41467-021-23545-7
  37. Mol Metab. 2021 May 24. pii: S2212-8778(21)00106-X. [Epub ahead of print] 101261
    Insulin action, type 2 diabetes, and branched-chain amino acids: a two-way street.
    Phillip J White, Robert W McGarrah, Mark A Herman, James R Bain, Svati H Shah, Christopher B Newgard.
      BACKGROUND: A strong association of obesity and insulin resistance with increased circulating levels of branched-chain and aromatic amino acids and decreased glycine levels has been recognized in human subjects for decades.SCOPE OF REVIEW: More recently, human metabolomic and genetic studies have confirmed and expanded upon these observations, accompanied by a surge in preclinical studies that have identified mechanisms involved in perturbation of amino acid homeostasis, how these events are connected to dysregulated glucose and lipid metabolism, and how elevations in branched-chain amino acids (BCAA) may participate in development of insulin resistance, type 2 diabetes (T2D) and other cardiometabolic diseases and conditions.
    MAJOR CONCLUSIONS: In human cohorts, BCAA and related metabolites are now well established as among the strongest biomarkers of obesity, insulin resistance, T2D, and cardiovascular diseases. Lowering of BCAA and branched-chain ketoacid (BCKA) levels by feeding of a BCAA-restricted diet or by activation of the rate limiting enzyme in BCAA catabolism, branched-chain ketoacid dehydrogenase (BCKDH) in rodent models of obesity has clear salutary effects on glucose and lipid homeostasis, but BCAA restriction has more modest effects in shorter-term studies in human T2D subjects. Feeding of rats with diets enriched in sucrose or fructose results in induction of the ChREBP transcription factor in liver to increase expression of the BCKDH kinase (BDK) and suppress expression of its phosphatase (PPM1K) resulting in inactivation of BCKDH and activation of the key lipogenic enzyme ATP-citrate lyase (ACLY). These and other emergent links between BCAA, glucose and lipid metabolism motivate ongoing studies of possible causal actions of BCAA and related metabolites in development of cardiometabolic diseases.
    Keywords:  Nutrition; branched-chain amino acids; insulin resistance; lipogenesis; metabolic diseases
    DOI:  https://doi.org/10.1016/j.molmet.2021.101261
  38. Cancer Res. 2021 May 28. pii: canres.0436.2021. [Epub ahead of print]
    Epigenetic induction of mitochondrial fission is required for maintenance of liver cancer initiating cells.
    Miaoling Tang, Meisongzhu Yang, Geyan Wu, Shuang Mo, Xingui Wu, Shuxia Zhang, Ruyuan Yu, Yameng Hu, Yingru Xu, Ziwen Li, Xinyi Liao, Jun Li, Libing Song.
      Mitochondrial dynamics play vital roles in the tumorigenicity and malignancy of various types of cancers by promoting the tumor-initiating potential of cancer cells, suggesting that targeting crucial factors that drive mitochondrial dynamics may lead to promising anticancer therapies. In the current study, we report that overexpression of mitochondrial fission factor (MFF), which is upregulated significantly in liver cancer initiating cells (LCIC), promotes mitochondrial fission and enhances stemness and tumor-initiating capability in non-LCICs. MFF-induced mitochondrial fission evoked mitophagy and asymmetric stem cell division and promoted a metabolic shift from oxidative phosphorylation to glycolysis that decreased mitochondrial reactive oxygen species (ROS) production, which prevented ROS-mediated degradation of the pluripotency transcription factor OCT4. CRISPR affinity purification in situ of regulatory elements (CAPTURE) showed that T-Box transcription factor 19 (TBX19), which is overexpressed uniquely in LCICs compared to non-LCICs and liver progenitor cells, forms a complex with PRMT1 on the MFF promoter in LCICs, eliciting epigenetic histone H4R3me2a/H3K9ac-mediated transactivation of MFF. Targeting PRMT1 using furamidine, a selective pharmacological inhibitor, suppressed TBX19-induced mitochondrial fission, leading to a profound loss of self-renewal potential and tumor-initiating capacity of LCICs. These findings unveil a novel mechanism underlying mitochondrial fission-mediated cancer stemness and suggest that regulation of mitochondrial fission via inhibition of PRMT1 may be an attractive therapeutic option for liver cancer treatment.
    DOI:  https://doi.org/10.1158/0008-5472.CAN-21-0436
  39. Genes Cells. 2021 May 25.
    VCP relocalization limits mitochondrial activity, GSH depletion, and ferroptosis during starvation in PC3 prostate cancer cells.
    Promise Ogor, Tomoki Yoshida, Masaaki Koike, Akira Kakizuka.
      During periods of crisis, cells must compensate to survive. To this end, cells may need to alter the subcellular localization of crucial proteins. Here, we show that during starvation, VCP, the most abundant soluble ATPase, relocalizes and forms aggregate-like structures at peri-nuclear regions in PC3 prostate cancer cells. This movement is associated with a lowered metabolic state, in which mitochondrial activity and ROS production are reduced. VCP appears to explicitly sense glutamine levels, as removal of glutamine from complete medium triggered VCP relocalization and its addition to starvation media blunted VCP relocalization. Cells cultured in Gln(+) starvation media exhibited uniformly distributed VCP in the cytoplasm (free VCP) and underwent ferroptotic cell death, which was associated with a decrease in GSH levels. Moreover, the addition of a VCP inhibitor, CB-5083, in starvation media prevented VCP relocalization and triggered ferroptotic cell death. Likewise, expression of GFP-fused VCP proteins, irrespective of ATPase activities, displayed free VCP and triggered cell death during starvation. These results indicate that free VCP is essential for the maintenance of mitochondrial function and that PC3 cells employ a strategy of VCP self-aggregation to suppress mitochondrial activity in order to escape cell death during starvation, a novel VCP-mediated survival mechanism.
    DOI:  https://doi.org/10.1111/gtc.12872
  40. Cell. 2021 May 27. pii: S0092-8674(21)00530-4. [Epub ahead of print]184(11): 2896-2910.e13
    Mitocytosis, a migrasome-mediated mitochondrial quality-control process.
    Haifeng Jiao, Dong Jiang, Xiaoyu Hu, Wanqing Du, Liangliang Ji, Yuzhuo Yang, Xiaopeng Li, Takami Sho, Xuan Wang, Ying Li, Yu-Ting Wu, Yau-Huei Wei, Xiaoyu Hu, Li Yu.
      Damaged mitochondria need to be cleared to maintain the quality of the mitochondrial pool. Here, we report mitocytosis, a migrasome-mediated mitochondrial quality-control process. We found that, upon exposure to mild mitochondrial stresses, damaged mitochondria are transported into migrasomes and subsequently disposed of from migrating cells. Mechanistically, mitocytosis requires positioning of damaged mitochondria at the cell periphery, which occurs because damaged mitochondria avoid binding to inward motor proteins. Functionally, mitocytosis plays an important role in maintaining mitochondrial quality. Enhanced mitocytosis protects cells from mitochondrial stressor-induced loss of mitochondrial membrane potential (MMP) and mitochondrial respiration; conversely, blocking mitocytosis causes loss of MMP and mitochondrial respiration under normal conditions. Physiologically, we demonstrate that mitocytosis is required for maintaining MMP and viability in neutrophils in vivo. We propose that mitocytosis is an important mitochondrial quality-control process in migrating cells, which couples mitochondrial homeostasis with cell migration.
    Keywords:  migrasome; mitochondrial quality control; mitochondrion; mitocytosis; mitosome
    DOI:  https://doi.org/10.1016/j.cell.2021.04.027
  41. Nat Metab. 2021 May;3(5): 714-727
    Single-cell metabolic imaging reveals a SLC2A3-dependent glycolytic burst in motile endothelial cells.
    David Wu, Devin L Harrison, Teodora Szasz, Chih-Fan Yeh, Tzu-Pin Shentu, Angelo Meliton, Ru-Ting Huang, Zhengjie Zhou, Gökhan M Mutlu, Jun Huang, Yun Fang.
      Single-cell motility is spatially heterogeneous and driven by metabolic energy. Directly linking cell motility to cell metabolism is technically challenging but biologically important. Here, we use single-cell metabolic imaging to measure glycolysis in individual endothelial cells with genetically encoded biosensors capable of deciphering metabolic heterogeneity at subcellular resolution. We show that cellular glycolysis fuels endothelial activation, migration and contraction and that sites of high lactate production colocalize with active cytoskeletal remodelling within an endothelial cell. Mechanistically, RhoA induces endothelial glycolysis for the phosphorylation of cofilin and myosin light chain in order to reorganize the cytoskeleton and thus control cell motility; RhoA activation triggers a glycolytic burst through the translocation of the glucose transporter SLC2A3/GLUT3 to fuel the cellular contractile machinery, as demonstrated across multiple endothelial cell types. Our data indicate that Rho-GTPase signalling coordinates energy metabolism with cytoskeleton remodelling to regulate endothelial cell motility.
    DOI:  https://doi.org/10.1038/s42255-021-00390-y
  42. Front Chem. 2021 ;9 672969
    A Quinol Anion as Catalytic Intermediate Coupling Proton Translocation With Electron Transfer in E. coli Respiratory Complex I.
    Franziska Nuber, Luca Mérono, Sabrina Oppermann, Johannes Schimpf, Daniel Wohlwend, Thorsten Friedrich.
      Energy-converting NADH:ubiquinone oxidoreductase, respiratory complex I, plays a major role in cellular energy metabolism. It couples NADH oxidation and quinone reduction with the translocation of protons across the membrane, thus contributing to the protonmotive force. Complex I has an overall L-shaped structure with a peripheral arm catalyzing electron transfer and a membrane arm engaged in proton translocation. Although both reactions are arranged spatially separated, they are tightly coupled by a mechanism that is not fully understood. Using redox-difference UV-vis spectroscopy, an unknown redox component was identified in Escherichia coli complex I as reported earlier. A comparison of its spectrum with those obtained for different quinone species indicates features of a quinol anion. The re-oxidation kinetics of the quinol anion intermediate is significantly slower in the D213GH variant that was previously shown to operate with disturbed quinone chemistry. Addition of the quinone-site inhibitor piericidin A led to strongly decreased absorption peaks in the difference spectrum. A hypothesis for a mechanism of proton-coupled electron transfer with the quinol anion as catalytically important intermediate in complex I is discussed.
    Keywords:  Escherichia coli; NADH dehydrogenase; bioenergetics; proton-coupled electron transfer; quinone chemistry; respiratory complex I
    DOI:  https://doi.org/10.3389/fchem.2021.672969
  43. Nat Commun. 2021 May 28. 12(1): 3210
    Mitochondrial targeted meganuclease as a platform to eliminate mutant mtDNA in vivo.
    Ugne Zekonyte, Sandra R Bacman, Jeff Smith, Wendy Shoop, Claudia V Pereira, Ginger Tomberlin, James Stewart, Derek Jantz, Carlos T Moraes.
      Diseases caused by heteroplasmic mitochondrial DNA mutations have no effective treatment or cure. In recent years, DNA editing enzymes were tested as tools to eliminate mutant mtDNA in heteroplasmic cells and tissues. Mitochondrial-targeted restriction endonucleases, ZFNs, and TALENs have been successful in shifting mtDNA heteroplasmy, but they all have drawbacks as gene therapy reagents, including: large size, heterodimeric nature, inability to distinguish single base changes, or low flexibility and effectiveness. Here we report the adaptation of a gene editing platform based on the I-CreI meganuclease known as ARCUS®. These mitochondrial-targeted meganucleases (mitoARCUS) have a relatively small size, are monomeric, and can recognize sequences differing by as little as one base pair. We show the development of a mitoARCUS specific for the mouse m.5024C>T mutation in the mt-tRNAAla gene and its delivery to mice intravenously using AAV9 as a vector. Liver and skeletal muscle show robust elimination of mutant mtDNA with concomitant restoration of mt-tRNAAla levels. We conclude that mitoARCUS is a potential powerful tool for the elimination of mutant mtDNA.
    DOI:  https://doi.org/10.1038/s41467-021-23561-7
  44. FEBS Open Bio. 2021 May 29.
    The mitochondrial fission factor FIS1 promotes stemness of human lung cancer stem cells via mitophagy.
    Doudou Liu, Zhiwei Sun, Ting Ye, Jingyuan Li, Bin Zeng, Qiting Zhao, Jianyu Wang, H Rosie Xing.
      Mitophagy, a form of autophagy, plays a role in cancer development, progression and recurrence. Cancer stem cells (CSCs) also play a key role in these processes, but it is unknown whether mitophagy can regulate the stemness of CSCs. Here, we employed the A549-SD human non-small cell lung adenocarcinoma CSC model that we have developed and characterized to investigate the effect of mitophagy on the stemness of CSCs. We observed a positive relationship between mitophagic activity and the stemness of lung CSCs. At the mechanistic level, our results suggest that augmentation of mitophagy in lung CSCs can be induced by FIS1 through mitochondrial fission. In addition, we assessed the clinical relevance of FIS1 in lung adenocarcinoma by using the TCGA database. An elevation in FIS1, when observed together with other prognostic markers for lung cancer progression, was found to correlate with shorter overall survival.
    Keywords:  FIS1; cancer stem cell; mitochondrial fission; mitophagy; stemness
    DOI:  https://doi.org/10.1002/2211-5463.13207
  45. Nat Metab. 2021 May;3(5): 618-635
    Mitohormesis reprogrammes macrophage metabolism to enforce tolerance.
    Greg A Timblin, Kevin M Tharp, Breanna Ford, Janet M Winchester, Jerome Wang, Stella Zhu, Rida I Khan, Shannon K Louie, Anthony T Iavarone, Johanna Ten Hoeve, Daniel K Nomura, Andreas Stahl, Kaoru Saijo.
      Macrophages generate mitochondrial reactive oxygen species and mitochondrial reactive electrophilic species as antimicrobials during Toll-like receptor (TLR)-dependent inflammatory responses. Whether mitochondrial stress caused by these molecules impacts macrophage function is unknown. Here, we demonstrate that both pharmacologically driven and lipopolysaccharide (LPS)-driven mitochondrial stress in macrophages triggers a stress response called mitohormesis. LPS-driven mitohormetic stress adaptations occur as macrophages transition from an LPS-responsive to LPS-tolerant state wherein stimulus-induced pro-inflammatory gene transcription is impaired, suggesting tolerance is a product of mitohormesis. Indeed, like LPS, hydroxyoestrogen-triggered mitohormesis suppresses mitochondrial oxidative metabolism and acetyl-CoA production needed for histone acetylation and pro-inflammatory gene transcription, and is sufficient to enforce an LPS-tolerant state. Thus, mitochondrial reactive oxygen species and mitochondrial reactive electrophilic species are TLR-dependent signalling molecules that trigger mitohormesis as a negative feedback mechanism to restrain inflammation via tolerance. Moreover, bypassing TLR signalling and pharmacologically triggering mitohormesis represents a new anti-inflammatory strategy that co-opts this stress response to impair epigenetic support of pro-inflammatory gene transcription by mitochondria.
    DOI:  https://doi.org/10.1038/s42255-021-00392-w
  46. Sci Rep. 2021 May 27. 11(1): 11185
    Small molecule inhibitors of the mitochondrial ClpXP protease possess cytostatic potential and re-sensitize chemo-resistant cancers.
    Martina Meßner, Melanie M Mandl, Mathias W Hackl, Till Reinhardt, Maximilian A Ardelt, Karolina Szczepanowska, Julian E Frädrich, Jens Waschke, Irmela Jeremias, Anja Fux, Matthias Stahl, Angelika M Vollmar, Stephan A Sieber, Johanna Pachmayr.
      The human mitochondrial ClpXP protease complex (HsClpXP) has recently attracted major attention as a target for novel anti-cancer therapies. Despite its important role in disease progression, the cellular role of HsClpXP is poorly characterized and only few small molecule inhibitors have been reported. Herein, we screened previously established S. aureus ClpXP inhibitors against the related human protease complex and identified potent small molecules against human ClpXP. The hit compounds showed anti-cancer activity in a panoply of leukemia, liver and breast cancer cell lines. We found that the bacterial ClpXP inhibitor 334 impairs the electron transport chain (ETC), enhances the production of mitochondrial reactive oxygen species (mtROS) and thereby promotes protein carbonylation, aberrant proteostasis and apoptosis. In addition, 334 induces cell death in re-isolated patient-derived xenograft (PDX) leukemia cells, potentiates the effect of DNA-damaging cytostatics and re-sensitizes resistant cancers to chemotherapy in non-apoptotic doses.
    DOI:  https://doi.org/10.1038/s41598-021-90801-7
  47. Sci Transl Med. 2021 May 26. pii: eabe8226. [Epub ahead of print]13(595):
    Hexose-6-phosphate dehydrogenase blockade reverses prostate cancer drug resistance in xenograft models by glucocorticoid inactivation.
    Jianneng Li, Michael Berk, Mohammad Alyamani, Navin Sabharwal, Christopher Goins, Joseph Alvarado, Mehdi Baratchian, Ziqi Zhu, Shaun Stauffer, Eric A Klein, Nima Sharifi.
      Prostate cancer resistance to next-generation hormonal treatment with enzalutamide is a major problem and eventuates into disease lethality. Biologically active glucocorticoids that stimulate glucocorticoid receptor (GR) have an 11β-OH moiety, and resistant tumors exhibit loss of 11β-HSD2, the oxidative (11β-OH → 11-keto) enzyme that normally inactivates glucocorticoids, allowing elevated tumor glucocorticoids to drive resistance by stimulating GR. Here, we show that up-regulation of hexose-6-phosphate dehydrogenase (H6PD) protein occurs in prostate cancer tissues of men treated with enzalutamide, human-derived cell lines, and patient-derived prostate tissues treated ex vivo with enzalutamide. Genetically silencing H6PD blocks NADPH generation, which inhibits the usual reductive directionality of 11β-HSD1, to effectively replace 11β-HSD2 function in human-derived cell line models, suppress the concentration of biologically active glucocorticoids in prostate cancer, and reverse enzalutamide resistance in mouse xenograft models. Similarly, pharmacologic blockade of H6PD with rucaparib normalizes tumor glucocorticoid metabolism in human cell lines and reinstates responsiveness to enzalutamide in mouse xenograft models. Our data show that blockade of H6PD, which is essential for glucocorticoid synthesis in humans, normalizes glucocorticoid metabolism and reverses enzalutamide resistance in mouse xenograft models. We credential H6PD as a pharmacologic vulnerability for treatment of next-generation androgen receptor antagonist-resistant prostate cancer by depleting tumor glucocorticoids.
    DOI:  https://doi.org/10.1126/scitranslmed.abe8226
  48. J Biol Chem. 2021 May 21. pii: S0021-9258(21)00623-2. [Epub ahead of print] 100825
    Perm1 promotes cardiomyocyte mitochondrial biogenesis and protects against hypoxia/reoxygenation-induced damage in mice.
    Yoshitake Cho, Shizuko Tachibana, Kayla Lam, Yoh Arita, Shamim Khosrowjerdi, Oliver Zhang, Alex Liang, Ruixia Li, Aleksander Andreyev, Anne N Murphy, Robert S Ross.
      Normal contractile function of the heart depends on a constant and reliable production of ATP by cardiomyocytes. Dysregulation of cardiac energy metabolism can result in immature heart development and disrupt the ability of the adult myocardium to adapt to stress, potentially leading to heart failure. Further, restoration of abnormal mitochondrial function can have beneficial effects on cardiac dysfunction. Previously, we identified a novel protein termed Perm1 (PGC-1 and ERR induced regulator, muscle 1) that is enriched in skeletal and cardiac-muscle mitochondria and transcriptionally regulated by PGC-1 (Peroxisome proliferator-activated receptor gamma coactivator 1) and ERR (Estrogen-related receptor). The role of Perm1 in the heart is poorly understood and was studied here. We utilized cell culture, mouse models and human tissue, to study its expression and transcriptional control, as well as its role in transcription of other factors. Critically, we tested Perm1's role in cardiomyocyte mitochondrial function and its ability to protect myocytes from stress-induced damage. Our studies show Perm1 expression increases throughout mouse cardiogenesis, demonstrate that Perm1 interacts with PGC-1α and enhances activation of PGC-1 and ERR, increases mitochondrial DNA copy number, and augments oxidative capacity in cultured neonatal mouse cardiomyocytes. Moreover, we found that Perm1 reduced cellular damage produced as a result of hypoxia and reoxygenation-induced stress and mitigated cell death of cardiomyocytes. Taken together, our results show that Perm1 promotes mitochondrial biogenesis in mouse cardiomyocytes. Future studies can assess the potential of Perm1 to be used as a novel therapeutic to restore cardiac dysfunction induced by ischemic injury.
    Keywords:  Perm1; cardiomyocytes; mitochondrial biogenesis; oxidative metabolism
    DOI:  https://doi.org/10.1016/j.jbc.2021.100825
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