bims-camemi Biomed News
on Mitochondrial metabolism in cancer
Issue of 2019–06–16
fifty papers selected by
Christian Frezza, , University of Cambridge, MRC Cancer Unit
  1. Connections between metabolism and epigenetics in Cancers.
  2. Cardiolipin-induced activation of pyruvate dehydrogenase links mitochondrial lipid biosynthesis to TCA cycle function.
  3. The Jekyll and Hyde character of RNase H1 and its multiple roles in mitochondrial DNA metabolism.
  4. Leptin promotes fatty acid oxidation and OXPHOS via the c-Myc/PGC-1 pathway in cancer cells.
  5. Non-oncogene Addiction to SIRT3 Plays a Critical Role in Lymphomagenesis.
  6. F-ATP Synthase and the Permeability Transition Pore: Fewer doubts, more certainties.
  7. PHB2 (prohibitin 2) promotes PINK1-PRKN/Parkin-dependent mitophagy by the PARL-PGAM5-PINK1 axis.
  8. Dynamic compartmentalization of purine nucleotide metabolic enzymes at leading edge in highly motile renal cell carcinoma.
  9. Loss of EZH2 Reprograms BCAA Metabolism to Drive Leukemic Transformation.
  10. Hypoxia and Metabolism in Metastasis.
  11. DRP1-mediated mitochondrial shape controls calcium homeostasis and muscle mass.
  12. Single organelle analysis to characterize mitochondrial function and crosstalk during viral infection.
  13. Consequences of inner mitochondrial membrane protein misfolding.
  14. Pyruvate Homeostasis as a Determinant of Parasite Growth and Metabolic Plasticity in Toxoplasma gondii.
  15. Role of coenzymes in Cancer Metabolism.
  16. Mitochondrial F-ATP Synthase and Its Transition into an Energy-Dissipating Molecular Machine.
  17. Retinoblastoma inactivation induces a protumoral microenvironment via enhanced CCL2 secretion.
  18. Metabolic Reprograming Via Silencing of Mitochondrial VDAC1 Expression Encourages Differentiation of Cancer Cells.
  19. Snail-Overexpression Induces Epithelial-mesenchymal Transition and Metabolic Reprogramming in Human Pancreatic Ductal Adenocarcinoma and Non-tumorigenic Ductal Cells.
  20. An acyl-CoA dehydrogenase microplate activity assay using recombinant porcine electron transfer flavoprotein.
  21. The Spatiotemporal Pattern and Intensity of p53 Activation Dictates Phenotypic Diversity in p53-Driven Developmental Syndromes.
  22. Mitochondrial pyruvate import is a metabolic vulnerability in androgen receptor-driven prostate cancer.
  23. Synergistic inhibition of tumor cell proliferation by metformin and mito-metformin in the presence of iron chelators.
  24. A chloroplast-localized mitochondrial calcium uniporter transduces osmotic stress in Arabidopsis.
  25. Inborn errors of mitochondrial acyl-coenzyme a metabolism: acyl-CoA biology meets the clinic.
  26. PDIA1/P4HB is required for efficient proinsulin maturation and ß cell health in response to diet induced obesity.
  27. The Mitochondrial Permeability Transition in Mitochondrial Disorders.
  28. The multifaceted contribution of α-ketoglutarate to tumor progression: An opportunity to exploit?
  29. The kynurenine pathway is essential for rhodoquinone biosynthesis in Caenorhabditis elegans.
  30. Identification of a Mitofusin specificity region that confers unique activities to Mfn1 and Mfn2.
  31. Cytosine-5 RNA methylation links protein synthesis to cell metabolism.
  32. 2-hydroxyglutarate inhibits MyoD-mediated differentiation by preventing H3K9 demethylation.
  33. Activation of P-TEFb by cAMP-PKA signaling in autosomal dominant polycystic kidney disease.
  34. 30 sweet years of GLUT4.
  35. Cytosolic phosphoenolpyruvate carboxykinase is expressed in α-cells from human and murine pancreas.
  36. Translatome analysis reveals altered serine and glycine metabolism in T-cell acute lymphoblastic leukemia cells.
  37. MITOL prevents ER stress-induced apoptosis by IRE1α ubiquitylation at ER-mitochondria contact sites.
  38. Dynamic genome-scale cell-specific metabolic models reveal novel inter-cellular and intra-cellular metabolic communications during ovarian follicle development.
  39. Metabolic regulation through the endosomal system.
  40. Enhanced acetylation of ATP-citrate lyase promotes the progression of nonalcoholic fatty liver disease.
  41. Determination of the metabolic index using the fluorescence lifetime of free and bound NADH in the phasor approach.
  42. Hsp90 middle domain phosphorylation initiates a complex conformational program to recruit the ATPase-stimulating cochaperone Aha1.
  43. High extracellular lactate causes reductive carboxylation in breast tissue cell lines grown under normoxic conditions.
  44. Gluconeogenesis of Cancer Cells Is Disrupted by Citrate.
  45. Tumor Heterogeneity Underlies Differential Cisplatin Sensitivity in Mouse Models of Small-Cell Lung Cancer.
  46. α-Ketoglutarate inhibits autophagy.
  47. Loss of 5-methylcytosine alters the biogenesis of vault-derived small RNAs to coordinate epidermal differentiation.
  48. Mitochondrial metabolism and glutamine are essential for mesoderm differentiation of human pluripotent stem cells.
  49. AMPK directly activates mTORC2 to promote cell survival during acute energetic stress.
  50. Metabolic influence on macrophage polarization and pathogenesis.
  1. Semin Cancer Biol. 2019 Jun 08. pii: S1044-579X(18)30082-8. [Epub ahead of print]
    Connections between metabolism and epigenetics in Cancers.
    Chitra Thakur, Fei Chen.
      In the past half century, our version on cancer, from tumor initiation, growth, to metastasis, is dominated by genetic mutation. The importance of metabolism and epigenetics was not recognized until most recently. Extensive cell proliferation is one of the hallmarks of cancers. To support the energetic and anabolic demands of enhanced proliferation, tumors reprogram the pathways of nutrient procurement and metabolism. In this context, a new link between metabolic alterations and cancer progression has been unraveled over the last decade by the studies conducted in the area of cancer cell metabolism. Cancer cells are known to alter their metabolic profile during the course of tumorigenesis and metastasis thereby exhibiting a tightly regulated program of metabolic plasticity. Noteworthy, certain metabolic alteration are known to occur at the epigenetic level, thus making epigenetics and metabolism highly interwoven in a reciprocal manner. Metabolites that are generated during metabolic pathways, such as in glycolytic cycle and oxidative phosphorylation, serve as cofactors or substrates for the enzymatic reactions that catalyze the epigenetic modifications and transcriptional regulation. Several studies also indicate that the epigenome is sensitive to cellular metabolism. Since many of the metabolic alterations and consequently aberrated epigenetic regulation are common to a wide range of cancer types, they serve as promising targets for anti-cancer therapies. Here we discuss the latest findings in cancer cell metabolism, elucidating the major anabolic, catabolic and energetic demands required for sustaining cancer growth, and the influence of altered metabolism on epigenetics and vice versa. A comprehensive research pertaining to metabolomic profiling and epigenome interactors/mediators in malignant neoplasias is imperative in deciphering the potential targets that can be exploited for the development of robust anti-cancer therapies.
    Keywords:  DNA and histone methylation; cancer cell metabolism; epigenetics; warburg effect
    DOI:  https://doi.org/10.1016/j.semcancer.2019.06.006
  2. J Biol Chem. 2019 Jun 11. pii: jbc.RA119.009037. [Epub ahead of print]
    Cardiolipin-induced activation of pyruvate dehydrogenase links mitochondrial lipid biosynthesis to TCA cycle function.
    Yiran Li, Wenjia Lou, Vaishnavi Raja, Simone Denis, Wenxi Yu, Michael W Schmidtke, Christian A Reynolds, Michael Schlame, Riekelt H Houtkooper, Miriam L Greenberg.
      Cardiolipin[MS1]  (CL) is the signature phospholipid of mitochondrial membranes. Although it has long been known that CL plays an important role in mitochondrial bioenergetics, recent evidence in the yeast model indicates that CL is also essential for intermediary metabolism. To gain insight into the function of CL in energy metabolism in mammalian cells, here we analyzed the metabolic flux of [U-13C]glucose in a mouse C2C12 myoblast cell line, TAZ-KO, which is CL-deficient because of a CRISPR/Cas9-mediated knockout of the CL-remodeling enzyme tafazzin (TAZ). TAZ-KO cells exhibited decreased flux of [U-13C]glucose to [13C]acetyl-CoA and M2 and M4 isotopomers of TCA cycle intermediates. Activity of pyruvate carboxylase (PC), the predominant enzyme for anaplerotic replenishing of the TCA cycle, was elevated in the TAZ-KO cells, which also exhibited increased sensitivity to the PC inhibitor phenylacetate. We attributed a decreased carbon flux from glucose to acetyl-CoA in the TAZ-KO cells to a ~50% decrease in pyruvate dehydrogenase (PDH) activity, which was observed in both TAZ-KO cells and cardiac tissue from TAZ-KO mice. Protein-lipid overlay experiments revealed that PDH binds to CL, and supplementing digitonin-solubilized TAZ-KO mitochondria with CL restored PDH activity to wildtype levels. Mitochondria from TAZ-KO cells exhibited an increase in phosphorylated PDH, levels of which were reduced in the presence of supplemented CL. These findings indicate that CL is required for optimal PDH activation, generation of acetyl-CoA, and TCA cycle function, findings that link the key mitochondrial lipid CL to TCA cycle function and energy metabolism.
    Keywords:  cardiolipin; mitochondria; pyruvate carboxylase (PC); pyruvate dehydrogenase complex (PDC); tricarboxylic acid cycle (TCA cycle) (Krebs cycle)
    DOI:  https://doi.org/10.1074/jbc.RA119.009037
  3. DNA Repair (Amst). 2019 Jun 04. pii: S1568-7864(19)30009-6. [Epub ahead of print]
    The Jekyll and Hyde character of RNase H1 and its multiple roles in mitochondrial DNA metabolism.
    Ian J Holt.
      The activity and specificity of ribonuclease H1, RNase H1, has been known for over half a century; like all enzymes in its class, it degrades RNA only when it is hybridized to DNA. However, the essential role of RNase H1 in mitochondrial DNA maintenance was not recognized until 2003, and empirical evidence that it is required to process RNA primers of mitochondrial DNA had to wait until 2015. In the same year, mutations in the RNASEH1 gene were linked to human mitochondrial diseases. The most recent studies suggest that in addition to primer-processing, RNase H1 determines the fate of R-loops, although not primarily those that might present an obstacle to DNA replication, but ones that contribute to the organization of mitochondrial DNA and the unusual mechanism of replication in mitochondria that utilizes transcripts for the strand-asynchronous mechanism of mitochondrial DNA replication. A full understanding of the role of RNase H1 in mtDNA metabolism will depend on further study, including careful consideration of its ability to stabilize, as well as to degrade RNA/DNA hybrids, and its regulation by oxidation or other mechanisms. Nevertheless, RNase H1 is already staking a strong claim to be the most versatile factor involved in propagating the DNA in the mitochondria.
    Keywords:  DNA replication; Mitochondrial DNA; Mitochondrial disease; Primers; R-loops; RNA/DNA hybrids; RNase H
    DOI:  https://doi.org/10.1016/j.dnarep.2019.06.001
  4. Acta Biochim Biophys Sin (Shanghai). 2019 Jun 12. pii: gmz058. [Epub ahead of print]
    Leptin promotes fatty acid oxidation and OXPHOS via the c-Myc/PGC-1 pathway in cancer cells.
    Qianqian Liu, Yang Sun, Zaiyi Fei, Zhibin Yang, Ke Duan, Jiaji Zi, Qinghua Cui, Min Yu, Wei Xiong.
      Alteration in cellular energy metabolism plays a critical role in the development and progression of cancer. Leptin is a hormone secreted by adipose tissue. Recent reports have shown that leptin can induce cancer cell proliferation and regulate cell energy metabolism, but the regulatory mechanism is still unclear. Here, we showed that leptin could promote cell proliferation and maintain high adenosine triphosphate levels in HCT116 and MCF-7 cells. The expression levels of carnitine palmitoyl transferase 1A (CPT1A), pyruvate dehydrogenase, succinate dehydrogenase subunit A and mitochondrial respiratory chain-associated proteins NADH dehydrogenase 1 (ND1), NADH:ubiquinone oxidoreductase subunit B8, and mitochondrial transcription factor A (TFAM) were distinctly increased in leptin-treated HCT116 and MCF-7 cells, while fatty acid synthase and lactate dehydrogenase expression were downregulated. Simultaneously, we found that c-Myc and peroxisome proliferator-activated receptor gamma co-activator 1 (PGC-1) protein expression levels were significantly increased. These results indicated that leptin boosted fatty acid β-oxidation and the tricarboxylic acid cycle, enhanced oxidative phosphorylation (OXPHOS) activity, and inhibited fatty acid synthesis and glycolysis in tumor cells. Gene transfection experiments revealed that leptin could induce the expression of c-Myc. Moreover, the expressions of PGC-1, CPT1A, and TFAM proteins were downregulated in HCT116 cells with low expression of c-Myc, and the expression levels of these proteins were increased in HCT116 cells overexpressing c-Myc. These findings suggest that leptin plays an important role in the regulation of energy metabolism in tumor cells. It may regulate fatty acid oxidation and OXPHOS of tumor cells by regulating the c-Myc/PGC-1 pathway. Targeting metabolic pathways for cancer treatment has been investigated as potential preventive or therapeutic methods. This study has important implications for the clinical therapy of tumor cell metabolism through hormone regulation.
    Keywords:  OXPHOS; c-Myc/PGC-1 pathway; cancer cells; fatty acid oxidation; leptin
    DOI:  https://doi.org/10.1093/abbs/gmz058
  5. Cancer Cell. 2019 Jun 10. pii: S1535-6108(19)30240-5. [Epub ahead of print]35(6): 916-931.e9
    Non-oncogene Addiction to SIRT3 Plays a Critical Role in Lymphomagenesis.
    Meng Li, Ying-Ling Chiang, Costas A Lyssiotis, Matthew R Teater, Jun Young Hong, Hao Shen, Ling Wang, Jing Hu, Hui Jing, Zhengming Chen, Neeraj Jain, Cihangir Duy, Sucharita J Mistry, Leandro Cerchietti, Justin R Cross, Lewis C Cantley, Michael R Green, Hening Lin, Ari M Melnick.
      Diffuse large B cell lymphomas (DLBCLs) are genetically heterogeneous and highly proliferative neoplasms derived from germinal center (GC) B cells. Here, we show that DLBCLs are dependent on mitochondrial lysine deacetylase SIRT3 for proliferation, survival, self-renewal, and tumor growth in vivo regardless of disease subtype and genetics. SIRT3 knockout attenuated B cell lymphomagenesis in VavP-Bcl2 mice without affecting normal GC formation. Mechanistically, SIRT3 depletion impaired glutamine flux to the TCA cycle via glutamate dehydrogenase and reduction in acetyl-CoA pools, which in turn induce autophagy and cell death. We developed a mitochondrial-targeted class I sirtuin inhibitor, YC8-02, which phenocopied the effects of SIRT3 depletion and killed DLBCL cells. SIRT3 is thus a metabolic non-oncogene addiction and therapeutic target for DLBCLs.
    Keywords:  DLBCL; GDH; SIRT3; TCA cycle; YC8-02 inhibitor; autophagy; cancer metabolism; glutaminolysis
    DOI:  https://doi.org/10.1016/j.ccell.2019.05.002
  6. FEBS Lett. 2019 Jun 14.
    F-ATP Synthase and the Permeability Transition Pore: Fewer doubts, more certainties.
    Michela Carraro, Vanessa Checchetto, Ildikó Szabò, Paolo Bernardi.
      Whether the mitochondrial permeability transition pore (PTP), also called mitochondrial megachannel (MMC), originates from the F-ATP synthase is a matter of controversy. This hypothesis is supported both by site-directed mutagenesis of specific residues of F-ATP synthase affecting regulation of the PTP/MMC and by deletion of specific subunits causing dramatic changes in channel conductance. In contrast, human cells lacking an assembled F-ATP synthase apparently display persistence of the PTP. We discuss recent data that shed new light on this controversy, supporting the conclusion that the PTP/MMC originates from a Ca2+ -dependent conformational change of F-ATP synthase allowing its reversible transformation into a high-conductance channel. This article is protected by copyright. All rights reserved.
    Keywords:  ATP synthase; Mitochondria; calcium; channel; cyclophilin; permeability transition
    DOI:  https://doi.org/10.1002/1873-3468.13485
  7. Autophagy. 2019 Jun 10.
    PHB2 (prohibitin 2) promotes PINK1-PRKN/Parkin-dependent mitophagy by the PARL-PGAM5-PINK1 axis.
    Chaojun Yan, Longlong Gong, Li Chen, Meng Xu, Hussein Abou-Hamdan, Mingliang Tang, Laurent Désaubry, Zhiyin Song.
      Mitophagy, which is a conserved cellular process for selectively removing damaged or unwanted mitochondria, is critical for mitochondrial quality control and the maintenance of normal cellular physiology. However, the precise mechanisms underlying mitophagy remain largely unknown. Prior studies on mitophagy focused on the events in the mitochondrial outer membrane. PHB2 (prohibitin 2), which is a highly conserved membrane scaffold protein, was recently identified as a novel inner membrane mitophagy receptor that mediates mitophagy. Here, we report a new signaling pathway for PHB2-mediated mitophagy. Upon mitochondrial membrane depolarization or misfolded protein aggregation, PHB2 depletion destabilizes PINK1 in the mitochondria, which blocks the mitochondrial recruitment of PRKN/Parkin, ubiquitin and OPTN (optineurin), leading to an inhibition of mitophagy. In addition, PHB2 overexpression directly induces PRKN recruitment to the mitochondria. Moreover, PHB2-mediated mitophagy is dependent on the mitochondrial inner membrane protease PARL, which interacts with PHB2 and is activated upon PHB2 depletion. Furthermore, PGAM5, which is processed by PARL, participates in PHB2-mediated PINK1 stabilization. Finally, a ligand of PHB proteins that we synthesized, called FL3, was found to strongly inhibit PHB2-mediated mitophagy and to effectively block cancer cell growth and energy production at nanomolar concentrations. Thus, our findings reveal that the PHB2-PARL-PGAM5-PINK1 axis is a novel pathway of PHB2-mediated mitophagy and that targeting PHB2 with the chemical compound FL3 is a promising strategy for cancer therapy.
    Keywords:  PARL; PGAM5; PHB2; PINK1-PRKN; mitophagy
    DOI:  https://doi.org/10.1080/15548627.2019.1628520
  8. Biochem Biophys Res Commun. 2019 Jun 10. pii: S0006-291X(19)31108-8. [Epub ahead of print]
    Dynamic compartmentalization of purine nucleotide metabolic enzymes at leading edge in highly motile renal cell carcinoma.
    Kara Wolfe, Satoshi Kofuji, Hirofumi Yoshino, Mika Sasaki, Koichi Okumura, Atsuo T Sasaki.
      Compartmentalization is vital for biological systems at multiple levels, including biochemical reactions in metabolism. Organelle-based compartments such as mitochondria and peroxisomes sequester the responsible enzymes and increase the efficiency of metabolism while simultaneously protecting the cell from dangerous intermediates, such as radical oxygen species. Recent studies show intracellular nucleotides, such as ATP and GTP, are heterogeneously distributed in cells with high concentrations at the lamellipodial and filopodial projections, or leading edge. However, the intracellular distribution of purine nucleotide enzymes remains unclear. Here, we report the enhanced localization of GTP-biosynthetic enzymes, including inosine monophosphate dehydrogenase (IMPDH isotype 1 and 2), GMP synthase (GMPS), guanylate kinase (GUK1) and nucleoside diphosphate kinase-A (NDPK-A) at the leading edge in renal cell carcinoma cells. They show significant co-localization at the membrane subdomain, and their co-localization pattern at the membrane is distinct from that of the cell body. While other purine nucleotide biosynthetic enzymes also show significant localization at the leading edge, their co-localization pattern with IMPDH is divergent. In contrast, a key glycolytic enzyme, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), predominantly localized in the cytoplasm. Mechanistically, we found that plasma membrane localization of IMPDH isozymes requires active actin polymerization. Our results demonstrate the formation of a discrete metabolic compartment for localized purine biosynthesis at the leading edge, which may promote localized nucleotide metabolism for cell migration and metastasis in cancers.
    Keywords:  GMPS; GUK1; IMPDH; Metabolic compartmentalization; Salvage purine synthesis; de novo purine synthesis
    DOI:  https://doi.org/10.1016/j.bbrc.2019.05.190
  9. Cancer Discov. 2019 Jun 12. pii: CD-19-0152. [Epub ahead of print]
    Loss of EZH2 Reprograms BCAA Metabolism to Drive Leukemic Transformation.
    Zhimin Gu, Yuxuan Liu, Feng Cai, McKenzie Patrick, Jakub Zmajkovic, Hui Cao, Yuannyu Zhang, Alpaslan Tasdogan, Mingyi Chen, Le Qi, Xin Liu, Kailong Li, Junhua Lyu, Kathryn E Dickerson, Weina Chen, Min Ni, Matthew E Merritt, Sean J Morrison, Radek C Skoda, Ralph J DeBerardinis, Jian Xu.
      Epigenetic gene regulation and metabolism are highly intertwined, yet little is known about whether altered epigenetics influence cellular metabolism during cancer progression. Here we show that EZH2 and NRasG12D mutations cooperatively induce progression of myeloproliferative neoplasms to highly penetrant, transplantable and lethal myeloid leukemias in mice. EZH1, an EZH2 homolog, is indispensable for EZH2-deficient leukemia-initiating cells and constitutes an epigenetic vulnerability. BCAT1, which catalyzes the reversible transamination of branched-chain amino acids (BCAAs), is repressed by EZH2 in normal hematopoiesis and aberrantly activated in EZH2-deficient myeloid neoplasms in mice and humans. BCAT1 reactivation cooperates with NRasG12D to sustain intracellular BCAA pools, resulting in enhanced mTOR signaling in EZH2-deficient leukemia cells. Genetic and pharmacological inhibition of BCAT1 selectively impairs EZH2-deficient leukemia-initiating cells and constitutes a metabolic vulnerability. Hence, epigenetic alterations rewire intracellular metabolism during leukemic transformation, causing epigenetic and metabolic vulnerabilities in cancer-initiating cells.
    DOI:  https://doi.org/10.1158/2159-8290.CD-19-0152
  10. Adv Exp Med Biol. 2019 ;1136 87-95
    Hypoxia and Metabolism in Metastasis.
    Tong Zhang, Caixia Suo, Chenyang Zheng, Huafeng Zhang.
      The hypoxic microenvironment is one of the major features of solid tumors, which regulates cell malignancy in multiple ways. As a response to hypoxia, a large number of target genes involved in cell growth, metabolism, metastasis and immunity are activated in cancer cells. Hypoxia-inducible factor 1 (HIF-1), as a heterodimeric DNA-binding complex, is comprised of a constitutively expressed HIF-1β subunit and an oxygen sensitive HIF-1α subunit, thus, adapts to decreased oxygen availability as a transcriptional factor. HIF-1 regulates many genes involved in tumorigenesis. Here, we focus on cancer cell metabolism and metastasis regulated by hypoxia.
    Keywords:  EMT; Glycogen synthesis; Glycolysis; HIF1; Hypoxia; Lipid metabolism; Metabolic enzymes; Metastasis; Metastatic niche; Mitochondria
    DOI:  https://doi.org/10.1007/978-3-030-12734-3_6
  11. Nat Commun. 2019 Jun 12. 10(1): 2576
    DRP1-mediated mitochondrial shape controls calcium homeostasis and muscle mass.
    Giulia Favaro, Vanina Romanello, Tatiana Varanita, Maria Andrea Desbats, Valeria Morbidoni, Caterina Tezze, Mattia Albiero, Marta Canato, Gaia Gherardi, Diego De Stefani, Cristina Mammucari, Bert Blaauw, Simona Boncompagni, Feliciano Protasi, Carlo Reggiani, Luca Scorrano, Leonardo Salviati, Marco Sandri.
      Mitochondrial quality control is essential in highly structured cells such as neurons and muscles. In skeletal muscle the mitochondrial fission proteins are reduced in different physiopathological conditions including ageing sarcopenia, cancer cachexia and chemotherapy-induced muscle wasting. However, whether mitochondrial fission is essential for muscle homeostasis is still unclear. Here we show that muscle-specific loss of the pro-fission dynamin related protein (DRP) 1 induces muscle wasting and weakness. Constitutive Drp1 ablation in muscles reduces growth and causes animal death while inducible deletion results in atrophy and degeneration. Drp1 deficient mitochondria are morphologically bigger and functionally abnormal. The dysfunctional mitochondria signals to the nucleus to induce the ubiquitin-proteasome system and an Unfolded Protein Response while the change of mitochondrial volume results in an increase of mitochondrial Ca2+ uptake and myofiber death. Our findings reveal that morphology of mitochondrial network is critical for several biological processes that control nuclear programs and Ca2+ handling.
    DOI:  https://doi.org/10.1038/s41467-019-10226-9
  12. Sci Rep. 2019 Jun 11. 9(1): 8492
    Single organelle analysis to characterize mitochondrial function and crosstalk during viral infection.
    Annika Schneider, Sandra Kurz, Katrin Manske, Marianne Janas, Mathias Heikenwälder, Thomas Misgeld, Michaela Aichler, Sebastian Felix Weissmann, Hans Zischka, Percy Knolle, Dirk Wohlleber.
      Mitochondria are key for cellular metabolism and signalling processes during viral infection. We report a methodology to analyse mitochondrial properties at the single-organelle level during viral infection using a recombinant adenovirus coding for a mitochondrial tracer protein for tagging and detection by multispectral flow cytometry. Resolution at the level of tagged individual mitochondria revealed changes in mitochondrial size, membrane potential and displayed a fragile phenotype during viral infection of cells. Thus, single-organelle and multi-parameter resolution allows to explore altered energy metabolism and antiviral defence by tagged mitochondria selectively in virus-infected cells and will be instrumental to identify viral immune escape and to develop and monitor novel mitochondrial-targeted therapies.
    DOI:  https://doi.org/10.1038/s41598-019-44922-9
  13. Mitochondrion. 2019 Jun 10. pii: S1567-7249(19)30086-8. [Epub ahead of print]
    Consequences of inner mitochondrial membrane protein misfolding.
    Liam P Coyne, Xin Jie Chen.
      Proteins embedded in the inner mitochondrial membrane (IMM) perform essential cellular functions. Maintaining the folding state of these proteins is therefore of the utmost importance, and this is ensured by IMM chaperones and proteases that refold and degrade unassembled and misfolded proteins. However, the physiological consequences specific to IMM protein misfolding remain obscure because deletion of these chaperones/proteases (the typical experimental strategy) often affects many mitochondrial processes other than protein folding and turnover. Thus, novel experimental systems are needed to evaluate the direct effects of misfolded protein on the membrane. Such a system has been developed in recent years. Studies suggest that numerous pathogenic mutations in isoform 1 of adenine nucleotide translocase (Ant1) cause its misfolding on the IMM. In this review, we first discuss potential mechanisms by which dominant Ant1 mutations may cause disease, highlighting IMM protein misfolding, per se, as a likely pathological factor. Then we discuss the intramitochondrial effects of Ant1 misfolding such as IMM proteostatic stress, respiratory chain dysfunction, and mtDNA instability. Finally, we summarize the mounting evidence that IMM proteostatic stress can perturb mitochondrial protein import to cause the toxic accumulation of mitochondrial proteins in the cytosol: a cell stress mechanism termed mitochondrial Precursor Overaccumulation Stress (mPOS).
    Keywords:  Ant1; Misfolding; Mitochondria; Mitochondrial carrier; mPOS
    DOI:  https://doi.org/10.1016/j.mito.2019.06.001
  14. MBio. 2019 Jun 11. pii: e00898-19. [Epub ahead of print]10(3):
    Pyruvate Homeostasis as a Determinant of Parasite Growth and Metabolic Plasticity in Toxoplasma gondii.
    Ningbo Xia, Shu Ye, Xiaohan Liang, Pu Chen, Yanqin Zhou, Rui Fang, Junlong Zhao, Nishith Gupta, Shuzhen Yang, Jing Yuan, Bang Shen.
      Toxoplasma gondii is a widespread intracellular pathogen infecting humans and a variety of animals. Previous studies have shown that Toxoplasma uses glucose and glutamine as the main carbon sources to support asexual reproduction, but neither nutrient is essential. Such metabolic flexibility may allow it to survive within diverse host cell types. Here, by focusing on the glycolytic enzyme pyruvate kinase (PYK) that converts phosphoenolpyruvate (PEP) into pyruvate, we found that Toxoplasma can also utilize lactate and alanine. We show that catabolism of all indicated carbon sources converges at pyruvate, and maintaining a constant pyruvate supply is critical to parasite growth. Toxoplasma expresses two PYKs: PYK1 in the cytosol and PYK2 in the apicoplast (a chloroplast relict). Genetic deletion of PYK2 did not noticeably affect parasite growth and virulence, which contrasts with the current model of carbon metabolism in the apicoplast. On the other hand, PYK1 was refractory to disruption. Conditional depletion of PYK1 resulted in global alteration of carbon metabolism, amylopectin accumulation, and reduced cellular ATP, leading to severe growth impairment. Notably, the attenuated growth of the PYK1-depleted mutant was partially rescued by lactate or alanine supplementation, and rescue by lactate required lactate dehydrogenase activity to convert it to pyruvate. Moreover, depletion of PYK1 in conjunction with PYK2 ablation led to accentuated loss of apicoplasts and complete growth arrest. Together, our results underline a critical role of pyruvate homeostasis in determining the metabolic flexibility and apicoplast maintenance, and they significantly extend our current understanding of carbon metabolism in T. gondii IMPORTANCE Toxoplasma gondii infects almost all warm-blooded animals, and metabolic flexibility is deemed critical for its successful parasitism in diverse hosts. Glucose and glutamine are the major carbon sources to support parasite growth. In this study, we found that Toxoplasma is also competent in utilizing lactate and alanine and, thus, exhibits exceptional metabolic versatility. Notably, all these nutrients need to be converted to pyruvate to fuel the lytic cycle, and achieving a continued pyruvate supply is vital to parasite survival and metabolic flexibility. Although pyruvate can be generated by two distinct pyruvate kinases, located in cytosol and apicoplast, respectively, the cytosolic enzyme is the main source of subcellular pyruvate, and cooperative usage of pyruvate among multiple organelles is critical for parasite growth and virulence. These findings expand our current understanding of carbon metabolism in Toxoplasma gondii and related parasites while providing a basis for designing novel antiparasitic interventions.
    Keywords:  Toxoplasma; apicoplast; lactate; metabolic flexibility; pyruvate kinase
    DOI:  https://doi.org/10.1128/mBio.00898-19
  15. Semin Cell Dev Biol. 2019 Jun 06. pii: S1084-9521(19)30070-9. [Epub ahead of print]
    Role of coenzymes in Cancer Metabolism.
    Maheshwor Thapa, Guido Dallmann.
      Cancer is a heterogeneous set of diseases characterized by the rewiring of cellular signaling and the reprogramming of metabolic pathways to sustain growth and proliferation. In past decades, studies were focused primarily on the genetic complexity of cancer. Recently, increasing number of studies have discovered several mutations among metabolic enzymes in different tumor cells. Most of the enzymes are regulated by coenzymes, organic cofactors, that function as intermediate carrier of electrons or functional groups that are transferred during the reaction. However, the precise role of cofactors is not well elucidated. In this review, we discuss several metabolic enzymes associated to cancer metabolism rewiring, whose inhibition may represent a therapeutic target. Such enzymes, upon expression or inhibition, may impact also the coenzymes levels, but only in few cases, it was possible to direct correlate coenzymes changes with a specific enzyme. In addition, we also summarize an up-to-date information on biological role of some coenzymes, preclinical and clinical studies, that have been carried out in various cancers and their outputs.
    Keywords:  cancer; coenzymes; mitochondria
    DOI:  https://doi.org/10.1016/j.semcdb.2019.05.027
  16. Oxid Med Cell Longev. 2019 ;2019 8743257
    Mitochondrial F-ATP Synthase and Its Transition into an Energy-Dissipating Molecular Machine.
    Giovanna Lippe, Gabriele Coluccino, Marco Zancani, Walter Baratta, Paola Crusiz.
      The mitochondrial F-ATP synthase is the principal energy-conserving nanomotor of cells that harnesses the proton motive force generated by the respiratory chain to make ATP from ADP and phosphate in a process known as oxidative phosphorylation. In the energy-converting membranes, F-ATP synthase is a multisubunit complex organized into a membrane-extrinsic F1 sector and a membrane-intrinsic FO domain, linked by central and peripheral stalks. Due to its essential role in the cellular metabolism, malfunction of F-ATP synthase has been associated with a variety of pathological conditions, and the enzyme is now considered as a promising drug target for multiple disease conditions and for the regulation of energy metabolism. We discuss structural and functional features of mitochondrial F-ATP synthase as well as several conditions that partially or fully inhibit the coupling between the F1 catalytic activities and the FO proton translocation, thus decreasing the cellular metabolic efficiency and transforming the enzyme into an energy-dissipating structure through molecular mechanisms that still remain to be defined.
    DOI:  https://doi.org/10.1155/2019/8743257
  17. Cancer Res. 2019 Jun 12. pii: canres.3604.2018. [Epub ahead of print]
    Retinoblastoma inactivation induces a protumoral microenvironment via enhanced CCL2 secretion.
    Fengkai Li, Shunsuke Kitajima, Susumu Kohno, Akiyo Yoshida, Shoichiro Tange, Soichiro Sasaki, Nobuhiro Okada, Yuuki Nishimoto, Hayato Muranaka, Naoko Nagatani, Misa Suzuki, Sayuri Masuda, Tran C Thai, Takumi Nishiuchi, Tomoaki Tanaka, David A Barbie, Naofumi Mukaida, Chiaki Takahashi.
      Cancer cell-intrinsic properties caused by oncogenic mutations have been well characterized, however, how specific oncogenes and tumor suppressors impact the tumor microenvironment (TME) is not well understood. Here, we present a novel non-cell-autonomous function of the retinoblastoma (RB) tumor suppressor in controlling the TME. RB inactivation stimulated tumor growth and neo-angiogenesis in a syngeneic and orthotropic murine soft tissue sarcoma model, which was associated with recruitment of tumor-associated macrophages (TAMs) and immunosuppressive cells such as Gr1+CD11b+ myeloid-derived suppressor cells (MDSCs) or Foxp3+ regulatory T cells (Tregs). Gene expression profiling and analysis of genetically engineered mouse models revealed that RB inactivation increased secretion of the chemoattractant CCL2. Furthermore, activation of the CCL2-CCR2 axis in the TME promoted tumor angiogenesis and recruitment of TAMs and MDSCs into the TME in several tumor types including sarcoma and breast cancer. Loss of RB increased fatty acid oxidation (FAO) by activating AMP-activated protein kinase (AMPK) which led to inactivation of Acetyl-CoA carboxylase (ACC) which suppresses FAO. This promoted mitochondrial superoxide production and JNK activation which enhanced CCL2 expression. These findings indicate that the CCL2-CCR2 axis could be an effective therapeutic target in RB-deficient tumors.
    DOI:  https://doi.org/10.1158/0008-5472.CAN-18-3604
  18. Mol Ther Nucleic Acids. 2019 May 18. pii: S2162-2531(19)30125-8. [Epub ahead of print]17 24-37
    Metabolic Reprograming Via Silencing of Mitochondrial VDAC1 Expression Encourages Differentiation of Cancer Cells.
    Tasleem Arif, Zohar Amsalem, Varda Shoshan-Barmatz.
      The mitochondrial gatekeeper voltage-dependent anion channel 1 (VDAC1) controls metabolic and energy cross-talk between mitochondria and the rest of the cell and is involved in mitochondria-mediated apoptosis. Here, we compared the effects of downregulated VDAC1 expression in the U-87MG glioblastoma, MDA-MB-231 triple-negative breast cancer (TNBC), and A549 lung cancer cell lines, using small interfering RNA (siRNA) specific to human VDAC1 (si-hVDAC1). The cells were subjected to si-hVDAC1 (50 nM) treatment for 5-20 days. Although VDAC1 silencing occurred within a day, the cells underwent reprograming with respect to rewiring metabolism, elimination of cancer stem cells (CSCs), and alteration of transcription factor (TF) expression and proteins associated with differentiation, with maximal changes being observed after 3 weeks of silencing VDAC1 expression. The differentiation into fewer tumorigenic cells may be associated with the elimination of CSCs. These alterations are interconnected, as protein up- or downregulation occurred simultaneously, starting 15-20 days after VDAC1 levels were first decreased. Moreover, the VDAC1 depletion-mediated effects on a network of key regulators of cell metabolism, CSCs, TFs, and other factors leading to differentiation are coordinated and are common to the glioblastoma multiforme (GBM) and lung and breast cancer cell lines, despite differing in origin and carried mutations. Thus, our study showed that VDAC1 depletion triggers reprograming of malignant cancer cells into terminally differentiated cells and that this may be a promising therapeutic approach for various cancers.
    Keywords:  VDAC1; cancer; cell differentiation; metabolism; mitochondria; siRNA
    DOI:  https://doi.org/10.1016/j.omtn.2019.05.003
  19. J Clin Med. 2019 Jun 08. pii: E822. [Epub ahead of print]8(6):
    Snail-Overexpression Induces Epithelial-mesenchymal Transition and Metabolic Reprogramming in Human Pancreatic Ductal Adenocarcinoma and Non-tumorigenic Ductal Cells.
    Menghan Liu, Sarah E Hancock, Ghazal Sultani, Brendan P Wilkins, Eileen Ding, Brenna Osborne, Lake-Ee Quek, Nigel Turner.
      The zinc finger transcription factor Snail is a known effector of epithelial-to-mesenchymal transition (EMT), a process that underlies the enhanced invasiveness and chemoresistance of common to cancerous cells. Induction of Snail-driven EMT has also been shown to drive a range of pro-survival metabolic adaptations in different cancers. In the present study, we sought to determine the specific role that Snail has in driving EMT and adaptive metabolic programming in pancreatic ductal adenocarcinoma (PDAC) by overexpressing Snail in a PDAC cell line, Panc1, and in immortalized, non-tumorigenic human pancreatic ductal epithelial (HPDE) cells. Snail overexpression was able to induce EMT in both pancreatic cell lines through suppression of epithelial markers and upregulation of mesenchymal markers alongside changes in cell morphology and enhanced migratory capacity. Snail-overexpressed pancreatic cells additionally displayed increased glucose uptake and lactate production with concomitant reduction in oxidative metabolism measurements. Snail overexpression reduced maximal respiration in both Panc1 and HPDE cells, with further reductions seen in ATP production, spare respiratory capacity and non-mitochondrial respiration in Snail overexpressing Panc1 cells. Accordingly, lower expression of mitochondrial electron transport chain proteins was observed with Snail overexpression, particularly within Panc1 cells. Modelling of 13C metabolite flux within both cell lines revealed decreased carbon flux from glucose in the TCA cycle in snai1-overexpressing Panc1 cells only. This work further highlights the role that Snail plays in EMT and demonstrates its specific effects on metabolic reprogramming of glucose metabolism in PDAC.
    Keywords:  SNA1; epithelial-mesenchymal transition; glucose metabolism; metabolomics; pancreatic adenocarcinoma.; tumor metabolism
    DOI:  https://doi.org/10.3390/jcm8060822
  20. Anal Biochem. 2019 Jun 10. pii: S0003-2697(19)30408-7. [Epub ahead of print]
    An acyl-CoA dehydrogenase microplate activity assay using recombinant porcine electron transfer flavoprotein.
    Yuxun Zhang, Al-Walid Mohsen, Catherine Kochersperger, Keaton Solo, Alexandra V Schmidt, Jerry Vockley, Eric S Goetzman.
      Acyl-CoA dehydrogenases (ACADs) play key roles in the mitochondrial catabolism of fatty acids and branched-chain amino acids. All nine characterized ACAD enzymes use electron transfer flavoprotein (ETF) as their redox partner. The gold standard for measuring ACAD activity is the anaerobic ETF fluorescence reduction assay, which follows the decrease of pig ETF fluorescence as it accepts electrons from an ACAD in vitro. Although first described 35 years ago, the assay has not been widely used due to the need to maintain an anaerobic assay environment and to purify ETF from pig liver mitochondria. Here, we present a method for expressing recombinant pig ETF in E coli and purifying it to homogeneity. The recombinant protein is virtually pure after one chromatography step, bears higher intrinsic fluorescence than the native enzyme, and provides enhanced activity in the ETF fluorescence reduction assay. Finally, we present a simplified protocol for removing molecular oxygen that allows adaption of the assay to a 96-well plate format. The availability of recombinant pig ETF and the microplate version of the ACAD activity assay will allow wide application of the assay for both basic research and clinical diagnostics.
    Keywords:  Acyl-CoA dehydrogenase; Electron transfer flavoprotein; Enzyme activity assay; Fatty acid oxidation; Mitochondria
    DOI:  https://doi.org/10.1016/j.ab.2019.06.003
  21. Dev Cell. 2019 May 28. pii: S1534-5807(19)30387-9. [Epub ahead of print]
    The Spatiotemporal Pattern and Intensity of p53 Activation Dictates Phenotypic Diversity in p53-Driven Developmental Syndromes.
    Margot E Bowen, Jacob McClendon, Hannah K Long, Aryo Sorayya, Jeanine L Van Nostrand, Joanna Wysocka, Laura D Attardi.
      Inappropriate activation of the p53 transcription factor contributes to numerous developmental syndromes characterized by distinct constellations of phenotypes. How p53 drives exquisitely specific sets of symptoms in diverse syndromes, however, remains enigmatic. Here, we deconvolute the basis of p53-driven developmental syndromes by leveraging an array of mouse strains to modulate the spatial expression pattern, temporal profile, and magnitude of p53 activation during embryogenesis. We demonstrate that inappropriate p53 activation in the neural crest, facial ectoderm, anterior heart field, and endothelium induces distinct spectra of phenotypes. Moreover, altering the timing and degree of p53 hyperactivation substantially affects the phenotypic outcomes. Phenotypes are associated with p53-driven cell-cycle arrest or apoptosis, depending on the cell type, with gene expression programs, rather than extent of mitochondrial priming, largely governing the specific response. Together, our findings provide a critical framework for decoding the role of p53 as a mediator of diverse developmental syndromes.
    Keywords:  Mdm2; apoptosis; cardiovascular; congenital defect; craniofacial; development; embryo; neural crest; p53; syndrome
    DOI:  https://doi.org/10.1016/j.devcel.2019.05.015
  22. Nat Metab. 2019 Jan;1(1): 70-85
    Mitochondrial pyruvate import is a metabolic vulnerability in androgen receptor-driven prostate cancer.
    David A Bader, Sean M Hartig, Vasanta Putluri, Christopher Foley, Mark P Hamilton, Eric A Smith, Pradip K Saha, Anil Panigrahi, Christopher Walker, Lin Zong, Heidi Martini-Stoica, Rui Chen, Kimal Rajapakshe, Cristian Coarfa, Arun Sreekumar, Nicholas Mitsiades, James A Bankson, Michael M Ittmann, Bert W O'Malley, Nagireddy Putluri, Sean E McGuire.
      Specific metabolic underpinnings of androgen receptor (AR)-driven growth in prostate adenocarcinoma (PCa) are largely undefined, hindering the development of strategies to leverage the metabolic dependencies of this disease when hormonal manipulations fail. Here we show that the mitochondrial pyruvate carrier (MPC), a critical metabolic conduit linking cytosolic and mitochondrial metabolism, is transcriptionally regulated by AR. Experimental MPC inhibition restricts proliferation and metabolic outputs of the citric acid cycle (TCA) including lipogenesis and oxidative phosphorylation in AR-driven PCa models. Mechanistically, metabolic disruption resulting from MPC inhibition activates the eIF2α/ATF4 integrated stress response (ISR). ISR signaling prevents cell cycle progression while coordinating salvage efforts, chiefly enhanced glutamine assimilation into the TCA, to regain metabolic homeostasis. We confirm that MPC function is operant in PCa tumors in-vivo using isotopomeric metabolic flux analysis. In turn, we apply a clinically viable small molecule targeting the MPC, MSDC0160, to pre-clinical PCa models and find that MPC inhibition suppresses tumor growth in hormone-responsive and castrate-resistant conditions. Collectively, our findings characterize the MPC as a tractable therapeutic target in AR-driven prostate tumors.
    DOI:  https://doi.org/10.1038/s42255-018-0002-y
  23. Oncotarget. 2019 May 28. 10(37): 3518-3532
    Synergistic inhibition of tumor cell proliferation by metformin and mito-metformin in the presence of iron chelators.
    Gang Cheng, Jacek Zielonka, Micael Hardy, Olivier Ouari, Christopher R Chitambar, Michael B Dwinell, Balaraman Kalyanaraman.
      We demonstrate that combined treatment with metformin (Met) or its mitochondria-targeted analog, mito-metformin (Mito-Met), and various iron chelators synergistically inhibits proliferation of pancreatic and triple-negative breast cancer cells. Previously, we reported that Met (at millimolar levels) and Mito-Met (at micromolar levels) inhibited pancreatic cancer cell proliferation. Inhibition of mitochondrial complex I and mitochondrial oxidative phosphorylation (OXPHOS) through stimulation of mitochondrial redox signaling was proposed as a key mechanism for decreased cancer cell proliferation. Because of its poor bioavailability, the use of Met as a "stand-alone" antitumor drug has been questioned. Iron chelators such as deferasirox (DFX) and dexrazoxane (DXR) exhibit antiproliferative effects in cancer cells. The potency of Met and Mito-Met was synergistically enhanced in the presence of iron chelators, DFX, N,N'-bis(2-hydroxyphenyl)ethylendiamine-N,N'-diacetic acid (HBED), and deferoxamine (DFO). Met, DXR (also ICRF-187), and DFO are FDA-approved drugs for treating diabetes, decreasing the incidence and severity of cardiotoxicity following chemotherapy, and mitigating iron toxicity, respectively. Thus, the synergistic antiproliferative effects of Met and Met analogs and iron chelators may have significant clinical relevance in cancer treatment. These findings may have implications in other OXPHOS-mediated cancer therapies.
    Keywords:  biguanide; metformin analogs; cancer cell proliferation; iron chelators; mitochondria-targeting agents
    DOI:  https://doi.org/10.18632/oncotarget.26943
  24. Nat Plants. 2019 Jun;5(6): 581-588
    A chloroplast-localized mitochondrial calcium uniporter transduces osmotic stress in Arabidopsis.
    Enrico Teardo, Luca Carraretto, Roberto Moscatiello, Enrico Cortese, Mattia Vicario, Margherita Festa, Lorenzo Maso, Sara De Bortoli, Tito Calì, Ute C Vothknecht, Elide Formentin, Laura Cendron, Lorella Navazio, Ildiko Szabo.
      Chloroplasts are integral to sensing biotic and abiotic stress in plants, but their role in transducing Ca2+-mediated stress signals remains poorly understood1,2. Here we identify cMCU, a member of the mitochondrial calcium uniporter (MCU) family, as an ion channel mediating Ca2+ flux into chloroplasts in vivo. Using a toolkit of aequorin reporters targeted to chloroplast stroma and the cytosol in cMCU wild-type and knockout lines, we provide evidence that stress-stimulus-specific Ca2+ dynamics in the chloroplast stroma correlate with expression of the channel. Fast downstream signalling events triggered by osmotic stress, involving activation of the mitogen-activated protein kinases (MAPK) MAPK3 and MAPK6, and the transcription factors MYB60 and ethylene-response factor 6 (ERF6), are influenced by cMCU activity. Relative to wild-type plants, cMCU knockouts display increased resistance to long-term water deficit and improved recovery on rewatering. Modulation of stromal Ca2+ in specific processing of stress signals identifies cMCU as a component of plant environmental sensing.
    DOI:  https://doi.org/10.1038/s41477-019-0434-8
  25. Mol Genet Metab. 2019 May 09. pii: S1096-7192(19)30097-6. [Epub ahead of print]
    Inborn errors of mitochondrial acyl-coenzyme a metabolism: acyl-CoA biology meets the clinic.
    Hao Yang, Chen Zhao, Marie-Christine Tang, Youlin Wang, Shu Pei Wang, Pierre Allard, Alexandra Furtos, Grant A Mitchell.
      The last decade saw major advances in understanding the metabolism of Coenzyme A (CoA) thioesters (acyl-CoAs) and related inborn errors (CoA metabolic diseases, CAMDs). For diagnosis, acylcarnitines and organic acids, both derived from acyl-CoAs, are excellent markers of most CAMDs. Clinically, each CAMD is unique but strikingly, three main patterns emerge: first, systemic decompensations with combinations of acidosis, ketosis, hypoglycemia, hyperammonemia and fatty liver; second, neurological episodes, particularly acute "stroke-like" episodes, often involving the basal ganglia but sometimes cerebral cortex, brainstem or optic nerves and third, especially in CAMDs of long chain fatty acyl-CoA metabolism, lipid myopathy, cardiomyopathy and arrhythmia. Some patients develop signs from more than one category. The pathophysiology of CAMDs is not precisely understood. Available data suggest that signs may result from CoA sequestration, toxicity and redistribution (CASTOR) in the mitochondrial matrix has been suggested to play a role. This predicts that most CAMDs cause deficiency of CoA, limiting mitochondrial energy production, and that toxic effects from the abnormal accumulation of acyl-CoAs and from extramitochondrial functions of acetyl-CoA may also contribute. Recent progress includes the following. (1) Direct measurements of tissue acyl-CoAs in mammalian models of CAMDs have been related to clinical features. (2) Inborn errors of CoA biosynthesis were shown to cause clinical changes similar to those of inborn errors of acyl-CoA degradation. (3) CoA levels in cells can be influenced pharmacologically. (4) Roles for acetyl-CoA are increasingly identified in all cell compartments. (5) Nonenzymatic acyl-CoA-mediated acylation of intracellular proteins occurs in mammalian tissues and is increased in CAMDs.
    DOI:  https://doi.org/10.1016/j.ymgme.2019.05.002
  26. Elife. 2019 Jun 11. pii: e44528. [Epub ahead of print]8
    PDIA1/P4HB is required for efficient proinsulin maturation and ß cell health in response to diet induced obesity.
    Insook Jang, Anita Pottekat, Juthakorn Poothong, Jing Yong, Jacqueline Lagunas-Acosta, Adriana Charbono, Zhouji Chen, Donalyn L Scheuner, Ming Liu, Pamela Itkin-Ansari, Peter Arvan, Randal J Kaufman.
      Regulated proinsulin biosynthesis, disulfide bond formation and ER redox homeostasis are essential to prevent Type two diabetes. In ß cells, protein disulfide isomerase A1 (PDIA1/P4HB), the most abundant ER oxidoreductase of over 17 members, can interact with proinsulin to influence disulfide maturation. Here we find Pdia1 is required for optimal insulin production under metabolic stress in vivo. ß cell-specific Pdia1 deletion in young high-fat diet fed mice or aged mice exacerbated glucose intolerance with inadequate insulinemia and increased the proinsulin/insulin ratio in both serum and islets compared to wildtype mice. Ultrastructural abnormalities in Pdia1-null ß cells include diminished insulin granule content, ER vesiculation and distention, mitochondrial swelling and nuclear condensation. Furthermore, Pdia1 deletion increased accumulation of disulfide-linked high molecular weight proinsulin complexes and islet vulnerability to oxidative stress. These findings demonstrate that PDIA1 contributes to oxidative maturation of proinsulin in the ER to support insulin production and ß cell health.
    Keywords:  PDIA1; beta cell function; cell biology; disulfide bond formation; glucose homeostasis; mouse; proinsulin maturation; type 2 diabetes
    DOI:  https://doi.org/10.7554/eLife.44528
  27. Oxid Med Cell Longev. 2019 ;2019 3403075
    The Mitochondrial Permeability Transition in Mitochondrial Disorders.
    Justina Šileikytė, Michael Forte.
      Mitochondrial permeability transition pore (PTP), a (patho)physiological phenomenon discovered over 40 years ago, is still not completely understood. PTP activation results in a formation of a nonspecific channel within the inner mitochondrial membrane with an exclusion size of 1.5 kDa. PTP openings can be transient and are thought to serve a physiological role to allow quick Ca2+ release and/or metabolite exchange between mitochondrial matrix and cytosol or long-lasting openings that are associated with pathological conditions. While matrix Ca2+ and oxidative stress are crucial in its activation, the consequence of prolonged PTP opening is dissipation of the inner mitochondrial membrane potential, cessation of ATP synthesis, bioenergetic crisis, and cell death-a primary characteristic of mitochondrial disorders. PTP involvement in mitochondrial and cellular demise in a variety of disease paradigms has been long appreciated, yet the exact molecular entity of the PTP and the development of potent and specific PTP inhibitors remain areas of active investigation. In this review, we will (i) summarize recent advances made in elucidating the molecular nature of the PTP focusing on evidence pointing to mitochondrial FoF1-ATP synthase, (ii) summarize studies aimed at discovering novel PTP inhibitors, and (iii) review data supporting compromised PTP activity in specific mitochondrial diseases.
    DOI:  https://doi.org/10.1155/2019/3403075
  28. Semin Cell Dev Biol. 2019 Jun 10. pii: S1084-9521(18)30171-X. [Epub ahead of print]
    The multifaceted contribution of α-ketoglutarate to tumor progression: An opportunity to exploit?
    Houda Abla, Manuela Sollazzo, Giuseppe Gasparre, Luisa Iommarini, Anna Maria Porcelli.
      The thriving field that constitutes cancer metabolism has unveiled some groundbreaking facts over the past two decades, at the heart of which is the TCA cycle and its intermediates. As such and besides its metabolic role, α-ketoglutarate was shown to withstand a wide range of physiological reactions from protection against oxidative stress, collagen and bone maintenance to development and immunity. Most importantly, it constitutes the rate-limiting substrate of 2-oxoglutarate-dependent dioxygenases family enzymes, which are involved in hypoxia sensing and in the shaping of cellular epigenetic landscape, two major drivers of oncogenic transformation. Based on literature reports, we hereby review the benefits of this metabolite as a possible novel adjuvant therapeutic opportunity to target tumor progression. This article is part of the special issue "Mitochondrial metabolic alterations in cancer cells and related therapeutic targets".
    Keywords:  Cancer metabolism; Drosophila melanogaster; Epigenetics; Hypoxia; Tumor progression; α-Ketoglutarate
    DOI:  https://doi.org/10.1016/j.semcdb.2019.05.031
  29. J Biol Chem. 2019 Jun 07. pii: jbc.AC119.009475. [Epub ahead of print]
    The kynurenine pathway is essential for rhodoquinone biosynthesis in Caenorhabditis elegans.
    Paloma M Roberts Buceta, Laura Romanelli-Cedrez, Shannon J Babcock, Helen Xun, Miranda L VonPaige, Thomas W Higley, Tyler D Schlatter, Dakota C Davis, Julia A Drexelius, John C Culver, Inés Carrera, Jennifer N Shepherd, Gustavo Salinas.
      A key metabolic adaptation of some species that face hypoxia as part of their life-cycle involves an alternative electron transport chain in which rhodoquinone (RQ) is required for fumarate reduction and ATP production. RQ biosynthesis in bacteria and protists requires ubiquinone (Q) as a precursor. In contrast, Q is not a precursor for RQ biosynthesis in animals such as parasitic helminths, and most details of this pathway have remained elusive. Here, we used Caenorhabditis elegans as a model animal to elucidate key steps in RQ biosynthesis. Using RNAi and a series of C. elegans mutants, we found that arylamine metabolites from the kynurenine pathway are essential precursors for RQ biosynthesis de novo Deletion of kynu-1, encoding a kynureninase that converts L-kynurenine (KYN) to anthranilic acid (AA), and 3-hydroxykynurenine (3HKYN) to 3-hydroxyanthranilic acid (3HAA), completely abolished RQ biosynthesis, but did not affect Q levels. Deletion of kmo-1, which encodes a kynurenine 3-monooxygenase that converts KYN to HKYN, drastically reduced RQ, but not Q levels. Knockdown of the Q biosynthetic genes coq-5 and coq-6 affected both Q and RQ levels, indicating that both biosynthetic pathways share common enzymes. Our study reveals that two pathways for RQ biosynthesis have independently evolved. Unlike in bacteria, where amination is the last step in RQ biosynthesis, in worms, the pathway begins with the arylamine precursor AA or 3HAA. Since RQ is absent in mammalian hosts of helminths, inhibition of RQ biosynthesis may have potential utility for targeting parasitic infections that cause important neglected tropical diseases.
    Keywords:  Caenorhabditis elegans (C. elegans); anthranilic acid; biosynthesis; electron transport; facultative anaerobe; helminths; hypoxia; kynureninase (kynu-1); rhodoquinone; ubiquinone
    DOI:  https://doi.org/10.1074/jbc.AC119.009475
  30. Mol Biol Cell. 2019 Jun 12. mbcE19050291
    Identification of a Mitofusin specificity region that confers unique activities to Mfn1 and Mfn2.
    S R Sloat, B N Whitley, E A Engelhart, S Hoppins.
      Mitochondrial structure can be maintained at steady state or modified in response to changes in cellular physiology. This is achieved by the coordinated regulation of dynamic properties including mitochondrial fusion, division and transport. Disease states, including neurodegeneration, are associated with defects in these processes. In vertebrates, two Mitofusin paralogs, Mfn1 and Mfn2, are required for efficient mitochondrial fusion. The Mitofusins share a high degree of homology and have very similar domain architecture, including an amino terminal GTPase domain and two extended helical bundles that are connected by flexible regions. Mfn1 and Mfn2 are non-redundant and are both required for mitochondrial outer membrane fusion. However, the molecular features that make these proteins functionally distinct are poorly defined. By engineering chimeric proteins composed of Mfn1 and Mfn2, we discovered a region that contributes to isoform-specific function (Mitofusin Isoform Specific Region - MISR). MISR confers unique fusion activity and Mitofusin specific nucleotide-dependent assembly properties. We propose that MISR functions in higher order oligomerization either directly, as an interaction interface, or indirectly through conformational changes.
    DOI:  https://doi.org/10.1091/mbc.E19-05-0291
  31. PLoS Biol. 2019 Jun 14. 17(6): e3000297
    Cytosine-5 RNA methylation links protein synthesis to cell metabolism.
    Nikoletta A Gkatza, Cecilia Castro, Robert F Harvey, Matthias Heiß, Martyna C Popis, Sandra Blanco, Susanne Bornelöv, Abdulrahim A Sajini, Joseph G Gleeson, Julian L Griffin, James A West, Stefanie Kellner, Anne E Willis, Sabine Dietmann, Michaela Frye.
      Posttranscriptional modifications in transfer RNA (tRNA) are often critical for normal development because they adapt protein synthesis rates to a dynamically changing microenvironment. However, the precise cellular mechanisms linking the extrinsic stimulus to the intrinsic RNA modification pathways remain largely unclear. Here, we identified the cytosine-5 RNA methyltransferase NSUN2 as a sensor for external stress stimuli. Exposure to oxidative stress efficiently repressed NSUN2, causing a reduction of methylation at specific tRNA sites. Using metabolic profiling, we showed that loss of tRNA methylation captured cells in a distinct catabolic state. Mechanistically, loss of NSUN2 altered the biogenesis of tRNA-derived noncoding fragments (tRFs) in response to stress, leading to impaired regulation of protein synthesis. The intracellular accumulation of a specific subset of tRFs correlated with the dynamic repression of global protein synthesis. Finally, NSUN2-driven RNA methylation was functionally required to adapt cell cycle progression to the early stress response. In summary, we revealed that changes in tRNA methylation profiles were sufficient to specify cellular metabolic states and efficiently adapt protein synthesis rates to cell stress.
    DOI:  https://doi.org/10.1371/journal.pbio.3000297
  32. Proc Natl Acad Sci U S A. 2019 Jun 10. pii: 201817662. [Epub ahead of print]
    2-hydroxyglutarate inhibits MyoD-mediated differentiation by preventing H3K9 demethylation.
    Juan-Manuel Schvartzman, Vincent P Reuter, Richard P Koche, Craig B Thompson.
      Oncogenic IDH1/2 mutations produce 2-hydroxyglutarate (2HG), resulting in competitive inhibition of DNA and protein demethylation. IDH-mutant cancer cells show an inability to differentiate but whether 2HG accumulation is sufficient to perturb differentiation directed by lineage-specifying transcription factors is unknown. A MyoD-driven model was used to study the role of IDH mutations in the differentiation of mesenchymal cells. The presence of 2HG produced by oncogenic IDH2 blocks the ability of MyoD to drive differentiation into myotubes. DNA 5mC hypermethylation is dispensable while H3K9 hypermethylation is required for this differentiation block. IDH2-R172K mutation results in H3K9 hypermethylation and impaired accessibility at myogenic chromatin regions but does not result in genome-wide decrease in accessibility. The results demonstrate the ability of the oncometabolite 2HG to block transcription factor-mediated differentiation in a molecularly defined system.
    Keywords:  2-hydroxyglutarate; differentiation; isocitrate dehydrogenase; myogenesis; sarcoma
    DOI:  https://doi.org/10.1073/pnas.1817662116
  33. Sci Adv. 2019 Jun;5(6): eaaw3593
    Activation of P-TEFb by cAMP-PKA signaling in autosomal dominant polycystic kidney disease.
    Yongzhan Sun, Zhiheng Liu, Xinyi Cao, Yi Lu, Zeyun Mi, Chaoran He, Jing Liu, Zhanye Zheng, Mulin Jun Li, Tiegang Li, Dechao Xu, Ming Wu, Ying Cao, Yuhao Li, Baoxue Yang, Changlin Mei, Lirong Zhang, Yupeng Chen.
      Positive transcription elongation factor b (P-TEFb) functions as a central regulator of transcription elongation. Activation of P-TEFb occurs through its dissociation from the transcriptionally inactive P-TEFb/HEXIM1/7SK snRNP complex. However, the mechanisms of signal-regulated P-TEFb activation and its roles in human diseases remain largely unknown. Here, we demonstrate that cAMP-PKA signaling disrupts the inactive P-TEFb/HEXIM1/7SK snRNP complex by PKA-mediated phosphorylation of HEXIM1 at serine-158. The cAMP pathway plays central roles in the development of autosomal dominant polycystic kidney disease (ADPKD), and we show that P-TEFb is hyperactivated in mouse and human ADPKD kidneys. Genetic activation of P-TEFb promotes cyst formation in a zebrafish ADPKD model, while pharmacological inhibition of P-TEFb attenuates cyst development by suppressing the pathological gene expression program in ADPKD mice. Our study therefore elucidates a mechanism by which P-TEFb activation by cAMP-PKA signaling promotes cystogenesis in ADPKD.
    DOI:  https://doi.org/10.1126/sciadv.aaw3593
  34. J Biol Chem. 2019 06 07. pii: jbc.REV119.008351. [Epub ahead of print]
    30 sweet years of GLUT4.
    Amira Klip, Timothy E McGraw, David E James.
      A pivotal metabolic function of insulin is the stimulation of glucose uptake into muscle and adipose tissues.  The discovery of the insulin-responsive glucose transporter type 4 (GLUT4) protein in 1988 inspired its molecular cloning in the following year. It also spurred numerous cellular mechanistic studies laying the foundations for how insulin regulates glucose uptake by muscle and fat cells. Here we reflect on the importance of the GLUT4 discovery and chronicle additional key findings made in the past 30 years. That exocytosis of a multi-spanning membrane protein regulates cellular glucose transport illuminated a novel adaptation of the secretory pathway, which is to transiently modulate the protein composition of the cellular plasma membrane. GLUT4 controls glucose transport into fat and muscle tissues in response to insulin and also into muscle during exercise. Thus, investigation of regulated GLUT4 trafficking provides a major means by which to map the essential signalling components that transmit the effects of insulin and exercise. Manipulation of the expression of GLUT4 or GLUT4-regulating molecules in mice has revealed the impact of glucose uptake on whole body metabolism. Remaining gaps in our understanding of GLUT4 function and regulation are highlighted here, along with opportunities for future discoveries and for the development of therapeutic approaches to manage metabolic disease.
    Keywords:  GLUT4 storage vesicle (GSV); diabetes; exercise; glucose metabolism; glucose transport; insulin; metabolic syndrome; signal transduction; solute carrer family 2 member 4 (SLC2A4)
    DOI:  https://doi.org/10.1074/jbc.REV119.008351
  35. J Cell Physiol. 2019 Jun 10.
    Cytosolic phosphoenolpyruvate carboxykinase is expressed in α-cells from human and murine pancreas.
    Francisco Westermeier, Todd Holyoak, Rodrigo Gatica, Fernando Martínez, Marianne Negrón, Alejandro J Yáñez, Daniel Nahmias, Francisco Nualart, Ian Burbulis, Romina Bertinat.
      The pancreatic islets of Langerhans, mainly formed by glucagon-producing α-cells and insulin-producing β-cells, are critical for glucose homeostasis. Insulin and glucagon oppositely modulate blood glucose levels in health, but a combined decline in insulin secretion together with increased glucagon secretion contribute to hyperglycemia in diabetes. Despite this bi-hormonal dysregulation, most studies have focused on insulin secretion and much less is known about glucagon secretion. Therefore, a deeper understanding of α-cell metabolism and glucagon secretion is of great interest. Here, we show that phosphoenolpyruvate carboxykinase (PCK1), an essential cataplerotic enzyme involved in metabolism and long considered to be absent from the pancreatic islet, is expressed in pancreatic α-cells of both murine and human. Furthermore, PCK1 transcription is induced by fasting and diabetes in rat pancreas, which indicates that the PCK1 activity is required for α-cell adaptation to different metabolic states. To our knowledge, this is the first evidence implicating PCK1 expression in α-cell metabolism.
    Keywords:  PCK1; diabetes; glucagon; insulin; pancreatic α-cell
    DOI:  https://doi.org/10.1002/jcp.28955
  36. Nat Commun. 2019 Jun 11. 10(1): 2542
    Translatome analysis reveals altered serine and glycine metabolism in T-cell acute lymphoblastic leukemia cells.
    Kim R Kampen, Laura Fancello, Tiziana Girardi, Gianmarco Rinaldi, Mélanie Planque, Sergey O Sulima, Fabricio Loayza-Puch, Benno Verbelen, Stijn Vereecke, Jelle Verbeeck, Joyce Op de Beeck, Jonathan Royaert, Pieter Vermeersch, David Cassiman, Jan Cools, Reuven Agami, Mark Fiers, Sarah-Maria Fendt, Kim De Keersmaecker.
      Somatic ribosomal protein mutations have recently been described in cancer, yet their impact on cellular transcription and translation remains poorly understood. Here, we integrate mRNA sequencing, ribosome footprinting, polysomal RNA sequencing and mass spectrometry datasets from a mouse lymphoid cell model to characterize the T-cell acute lymphoblastic leukemia (T-ALL) associated ribosomal RPL10 R98S mutation. Surprisingly, RPL10 R98S induces changes in protein levels primarily through transcriptional rather than translation efficiency changes. Phosphoserine phosphatase (PSPH), encoding a key serine biosynthesis enzyme, was the only gene with elevated transcription and translation leading to protein overexpression. PSPH upregulation is a general phenomenon in T-ALL patient samples, associated with elevated serine and glycine levels in xenograft mice. Reduction of PSPH expression suppresses proliferation of T-ALL cell lines and their capacity to expand in mice. We identify ribosomal mutation driven induction of serine biosynthesis and provide evidence supporting dependence of T-ALL cells on PSPH.
    DOI:  https://doi.org/10.1038/s41467-019-10508-2
  37. EMBO J. 2019 Jun 13. pii: e100999. [Epub ahead of print]
    MITOL prevents ER stress-induced apoptosis by IRE1α ubiquitylation at ER-mitochondria contact sites.
    Keisuke Takeda, Shun Nagashima, Isshin Shiiba, Aoi Uda, Takeshi Tokuyama, Naoki Ito, Toshifumi Fukuda, Nobuko Matsushita, Satoshi Ishido, Takao Iwawaki, Takashi Uehara, Ryoko Inatome, Shigeru Yanagi.
      Unresolved endoplasmic reticulum (ER) stress shifts the unfolded protein response signaling from cell survival to cell death, although the switching mechanism remains unclear. Here, we report that mitochondrial ubiquitin ligase (MITOL/MARCH5) inhibits ER stress-induced apoptosis through ubiquitylation of IRE1α at the mitochondria-associated ER membrane (MAM). MITOL promotes K63-linked chain ubiquitination of IRE1α at lysine 481 (K481), thereby preventing hyper-oligomerization of IRE1α and regulated IRE1α-dependent decay (RIDD). Therefore, under ER stress, MITOL depletion or the IRE1α mutant (K481R) allows for IRE1α hyper-oligomerization and enhances RIDD activity, resulting in apoptosis. Similarly, in the spinal cord of MITOL-deficient mice, ER stress enhances RIDD activity and subsequent apoptosis. Notably, unresolved ER stress attenuates IRE1α ubiquitylation, suggesting that this directs the apoptotic switch of IRE1α signaling. Our findings suggest that mitochondria regulate cell fate under ER stress through IRE1α ubiquitylation by MITOL at the MAM.
    Keywords:  IRE1α; apoptosis; mitochondrial E3 ligase MITOL/MARCH5; mitochondria‐associated ER membrane; unfolded protein response
    DOI:  https://doi.org/10.15252/embj.2018100999
  38. BMC Bioinformatics. 2019 Jun 10. 20(1): 307
    Dynamic genome-scale cell-specific metabolic models reveal novel inter-cellular and intra-cellular metabolic communications during ovarian follicle development.
    Beatriz Peñalver Bernabé, Ines Thiele, Eugene Galdones, Anaar Siletz, Sriram Chandrasekaran, Teresa K Woodruff, Linda J Broadbelt, Lonnie D Shea.
       BACKGROUND: The maturation of the female germ cell, the oocyte, requires the synthesis and storing of all the necessary metabolites to support multiple divisions after fertilization. Oocyte maturation is only possible in the presence of surrounding, diverse, and changing layers of somatic cells. Our understanding of metabolic interactions between the oocyte and somatic cells has been limited due to dynamic nature of ovarian follicle development, thus warranting a systems approach.
    RESULTS: Here, we developed a genome-scale metabolic model of the mouse ovarian follicle. This model was constructed using an updated mouse general metabolic model (Mouse Recon 2) and contains several key ovarian follicle development metabolic pathways. We used this model to characterize the changes in the metabolism of each follicular cell type (i.e., oocyte, granulosa cells, including cumulus and mural cells), during ovarian follicle development in vivo. Using this model, we predicted major metabolic pathways that are differentially active across multiple follicle stages. We identified a set of possible secreted and consumed metabolites that could potentially serve as biomarkers for monitoring follicle development, as well as metabolites for addition to in vitro culture media that support the growth and maturation of primordial follicles.
    CONCLUSIONS: Our systems approach to model follicle metabolism can guide future experimental studies to validate the model results and improve oocyte maturation approaches and support growth of primordial follicles in vitro.
    Keywords:  Cell-type specific metabolic models; Genome-scale modeling; Metabolic communities; Metabolism; Ovarian follicle development; Secreted metabolites
    DOI:  https://doi.org/10.1186/s12859-019-2825-2
  39. Traffic. 2019 Jun 08.
    Metabolic regulation through the endosomal system.
    Jerome Gilleron, Jantje M Gerdes, Anja Zeigerer.
      The endosomal system plays an essential role in cell homeostasis by controlling cellular signaling, nutrient sensing, cell polarity and cell migration. However, its place in the regulation of tissue, organ and whole body physiology is less well understood. Recent studies have revealed an important role for the endosomal system in regulating glucose and lipid homeostasis, with implications for metabolic disorders such as type-2 diabetes, hypercholesterolemia and non-alcoholic fatty liver disease. By taking insights from in vitro studies of endocytosis and exploring their effects on metabolism, we can begin to connect the fields of endosomal transport and metabolic homeostasis. In this review, we explore current understanding of how the endosomal system influences the systemic regulation of glucose and lipid metabolism in mice and humans. We highlight exciting new insights that help translate findings from single cells to a wider physiological level and open up new directions for endosomal research. This article is protected by copyright. All rights reserved.
    Keywords:  diabetes; endocytosis; fatty liver disease; glucose and lipid metabolism; metabolic signaling; nutrient transport
    DOI:  https://doi.org/10.1111/tra.12670
  40. J Biol Chem. 2019 Jun 13. pii: jbc.RA119.008708. [Epub ahead of print]
    Enhanced acetylation of ATP-citrate lyase promotes the progression of nonalcoholic fatty liver disease.
    Liang Guo, Ying-Ying Guo, Bai-Yu Li, Wan-Qiu Peng, Xin-Xia Chang, Xin Gao, Qi-Qun Tang.
      Hepatic steatosis is a hallmark of nonalcoholic fatty liver disease (NAFLD) and is promoted by dysregulated de novo lipogenesis. ATP-citrate lyase (ACLY) is a crucial lipogenic enzyme that is up-regulated in individuals with NAFLD. A previous study has shown that acetylation of ACLY at Lys-540, Lys-546, and Lys-554 (ACLY-3K) increases ACLY protein stability by antagonizing its ubiquitylation, thereby promoting lipid synthesis and cell proliferation in lung cancer cells. But the functional importance of this regulatory mechanism in other cellular or tissue contexts or under other pathophysiological conditions awaits further investigation. Here, we show that ACLY-3K acetylation also promotes ACLY protein stability in AML12 cells, a mouse hepatocyte cell line, and found that the deacetylase sirtuin 2 (SIRT2) deacetylates ACLY-3K and destabilizes ACLY in these cells. Of note, the livers of mice and humans with NAFLD had increased ACLY protein and ACLY-3K acetylation levels and decreased SIRT2 protein levels. Mimicking ACLY-3K acetylation by replacing the three lysines with three glutamines (ACLY-3KQ variant) promoted lipid accumulation both in high glucose-treated AML12 cells and in the livers of high-fat/high-sucrose (HF/HS) diet-fed mice. Moreover, overexpressing SIRT2 in AML12 cells inhibited lipid accumulation, which was more efficiently reversed by overexpressing the ACLY-3KQ variant than by overexpressing wildtype ACLY. Additionally, hepatic SIRT2 overexpression decreased ACLY-3K acetylation and its protein level and alleviated hepatic steatosis in HF/HS diet-fed mice. Our findings reveal a posttranscriptional mechanism underlying the up-regulation of hepatic ACLY in NAFLD and suggest that the SIRT2/ACLY axis is involved in NAFLD progression.
    Keywords:  ATP-citrate lyase (ACLY); acetylation; deacetylase; hepatic steatosis; hepatocyte; lipid metabolism; lipogenesis; metabolic disorder; nonalcoholic fatty liver disease (NAFLD); protein stability; sirtuin
    DOI:  https://doi.org/10.1074/jbc.RA119.008708
  41. J Biophotonics. 2019 Jun 13. e201900156
    Determination of the metabolic index using the fluorescence lifetime of free and bound NADH in the phasor approach.
    Suman Ranjit, Leonel Malacrida, Milka Stakic, Enrico Gratton.
      The fluorescence lifetime of nicotinamide adenine dinucleotide (NADH) is commonly used in conjunction with the phasor approach as a molecular biomarker to provide information on cellular metabolism of autofluorescence imaging of cells and tissue. However in the phasor approach, the bound and free lifetime defining the phasor metabolic trajectory is a subject of debate. NADH increases the fluorescence lifetime when bound to an enzyme, in contrast to the short multiexponential lifetime displayed by NADH in solution. The extent of fluorescence lifetime increase depends on the enzyme to which NADH is bound. With proper preparation of lactate dehydrogenase (LDH) using oxalic acid as an allosteric factor, bound NADH to LDH has a lifetime of 3.4 ns and is positioned on the universal semi-circle of the phasor plot, inferring a mono-exponential lifetime for this species. Surprisingly, measurements in the cellular environments with different metabolic states show a linear trajectory between free NADH at about 0.37 ns and bound NADH at 3.4ns. These observations support that in a cellular environment a 3.4 ns value could be used for bound NADH lifetime. The phasor analysis of many cell types shows a linear combination of fractional contributions of free and bound species NADH. This article is protected by copyright. All rights reserved.
    Keywords:  Autofluorescence; FLIM; NADH; Phasor; TCSPC; lifetime
    DOI:  https://doi.org/10.1002/jbio.201900156
  42. Nat Commun. 2019 Jun 12. 10(1): 2574
    Hsp90 middle domain phosphorylation initiates a complex conformational program to recruit the ATPase-stimulating cochaperone Aha1.
    Wanping Xu, Kristin Beebe, Juan D Chavez, Marta Boysen, YinYing Lu, Abbey D Zuehlke, Dimitra Keramisanou, Jane B Trepel, Christosomos Prodromou, Matthias P Mayer, James E Bruce, Ioannis Gelis, Len Neckers.
      Complex conformational dynamics are essential for function of the dimeric molecular chaperone heat shock protein 90 (Hsp90), including transient, ATP-biased N-domain dimerization that is necessary to attain ATPase competence. The intrinsic, but weak, ATP hydrolyzing activity of human Hsp90 is markedly enhanced by the co-chaperone Aha1. However, the cellular concentration of Aha1 is substoichiometric relative to Hsp90. Here we report that initial recruitment of this cochaperone to Hsp90 is markedly enhanced by phosphorylation of a highly conserved tyrosine (Y313 in Hsp90α) in the Hsp90 middle domain. Importantly, phosphomimetic mutation of Y313 promotes formation of a transient complex in which both N- and C-domains of Aha1 bind to distinct surfaces of the middle domains of opposing Hsp90 protomers prior to ATP-directed N-domain dimerization. Thus, Y313 represents a phosphorylation-sensitive conformational switch, engaged early after client loading, that affects both local and long-range conformational dynamics to facilitate initial recruitment of Aha1 to Hsp90.
    DOI:  https://doi.org/10.1038/s41467-019-10463-y
  43. PLoS One. 2019 ;14(6): e0213419
    High extracellular lactate causes reductive carboxylation in breast tissue cell lines grown under normoxic conditions.
    Arthur Nathan Brodsky, Daniel C Odenwelder, Sarah W Harcum.
      In cancer tumors, lactate accumulation was initially attributed to high glucose consumption associated with the Warburg Effect. Now it is evident that lactate can also serve as an energy source in cancer cell metabolism. Additionally, lactate has been shown to promote metastasis, generate gene expression patterns in cancer cells consistent with "cancer stem cell" phenotypes, and result in treatment resistant tumors. Therefore, the goal of this work was to quantify the impact of lactate on metabolism in three breast cell lines (one normal and two breast cancer cell lines-MCF 10A, MCF7, and MDA-MB-231), in order to better understand the role lactate may have in different disease cell types. Parallel labeling metabolic flux analysis (13C-MFA) was used to quantify the intracellular fluxes under normal and high extracellular lactate culture conditions. Additionally, high extracellular lactate cultures were labelled in parallel with [U-13C] lactate, which provided qualitative information regarding the lactate uptake and metabolism. The 13C-MFA model, which incorporated the measured extracellular fluxes and the parallel labeling mass isotopomer distributions (MIDs) for five glycolysis, four tricarboxylic acid cycle (TCA), and three intracellular amino acid metabolites, predicted lower glycolysis fluxes in the high lactate cultures. All three cell lines experienced reductive carboxylation of glutamine to citrate in the TCA cycle as a result of high extracellular lactate. Reductive carboxylation previously has been observed under hypoxia and other mitochondrial stresses, whereas these cultures were grown aerobically. In addition, this is the first study to investigate the intracellular metabolic responses of different stages of breast cancer progression to high lactate exposure. These results provide insight into the role lactate accumulation has on metabolic reaction distributions in the different disease cell types while the cells are still proliferating in lactate concentrations that do not significantly decrease exponential growth rates.
    DOI:  https://doi.org/10.1371/journal.pone.0213419
  44. Trends Cancer. 2019 May;pii: S2405-8033(19)30066-4. [Epub ahead of print]5(5): 265-266
    Gluconeogenesis of Cancer Cells Is Disrupted by Citrate.
    Philippe Icard, Zherui Wu, Marco Alifano, Ludovic Fournel.
      
    Keywords:  FBPase; PFK; Warburg effect; cancer metabolism; citrate; gluconeogenesis
    DOI:  https://doi.org/10.1016/j.trecan.2019.03.002
  45. Cell Rep. 2019 Jun 11. pii: S2211-1247(19)30686-2. [Epub ahead of print]27(11): 3345-3358.e4
    Tumor Heterogeneity Underlies Differential Cisplatin Sensitivity in Mouse Models of Small-Cell Lung Cancer.
    Franziska Böttger, Ekaterina A Semenova, Ji-Ying Song, Giustina Ferone, Jan van der Vliet, Miranda Cozijnsen, Rajith Bhaskaran, Lorenzo Bombardelli, Sander R Piersma, Thang V Pham, Connie R Jimenez, Anton Berns.
      Small-cell lung cancer is the most aggressive type of lung cancer, characterized by a remarkable response to chemotherapy followed by development of resistance. Here, we describe SCLC subtypes in Mycl- and Nfib-driven GEMM that include CDH1-high peripheral primary tumor lesions and CDH1-negative, aggressive intrapulmonary metastases. Cisplatin treatment preferentially eliminates the latter, thus revealing a striking differential response. Using a combined transcriptomic and proteomic approach, we find a marked reduction in proliferation and metabolic rewiring following cisplatin treatment and present evidence for a distinctive metabolic and structural profile defining intrinsically resistant populations. This offers perspectives for effective combination therapies that might also hold promise for treating human SCLC, given the very similar response of both mouse and human SCLC to cisplatin.
    Keywords:  RNA-seq; SCLC; chemotherapy; cisplatin; mass spectrometry; mouse models; proteomics; transcriptomics; tumor heterogeneity
    DOI:  https://doi.org/10.1016/j.celrep.2019.05.057
  46. Aging (Albany NY). 2019 Jun 07.
    α-Ketoglutarate inhibits autophagy.
    Elisa Elena Baracco, Francesca Castoldi, Sylvère Durand, David P Enot, Jelena Tadic, Katharina Kainz, Frank Madeo, Alexis Chery, Valentina Izzo, Maria Chiara Maiuri, Federico Pietrocola, Guido Kroemer.
      The metabolite α-ketoglutarate is membrane-impermeable, meaning that it is usually added to cells in the form of esters such as dimethyl -ketoglutarate (DMKG), trifluoromethylbenzyl α-ketoglutarate (TFMKG) and octyl α-ketoglutarate (O-KG). Once these compounds cross the plasma membrane, they are hydrolyzed by esterases to generate α-ketoglutarate, which remains trapped within cells. Here, we systematically compared DMKG, TFMKG and O-KG for their metabolic and functional effects. All three compounds similarly increased the intracellular levels of α-ketoglutarate, yet each of them had multiple effects on other metabolites that were not shared among the three agents, as determined by mass spectrometric metabolomics. While all three compounds reduced autophagy induced by culture in nutrient-free conditions, TFMKG and O-KG (but not DMKG) caused an increase in baseline autophagy in cells cultured in complete medium. O-KG (but neither DMKG nor TFMK) inhibited oxidative phosphorylation and exhibited cellular toxicity. Altogether, these results support the idea that intracellular α-ketoglutarate inhibits starvation-induced autophagy and that it has no direct respiration-inhibitory effect.
    Keywords:  Krebs cycle; acetyl CoA; aging; cell death; metabolomics; mitochondria
    DOI:  https://doi.org/10.18632/aging.102001
  47. Nat Commun. 2019 Jun 11. 10(1): 2550
    Loss of 5-methylcytosine alters the biogenesis of vault-derived small RNAs to coordinate epidermal differentiation.
    Abdulrahim A Sajini, Nila Roy Choudhury, Rebecca E Wagner, Susanne Bornelöv, Tommaso Selmi, Christos Spanos, Sabine Dietmann, Juri Rappsilber, Gracjan Michlewski, Michaela Frye.
      The presence and absence of RNA modifications regulates RNA metabolism by modulating the binding of writer, reader, and eraser proteins. For 5-methylcytosine (m5C) however, it is largely unknown how it recruits or repels RNA-binding proteins. Here, we decipher the consequences of m5C deposition into the abundant non-coding vault RNA VTRNA1.1. Methylation of cytosine 69 in VTRNA1.1 occurs frequently in human cells, is exclusively mediated by NSUN2, and determines the processing of VTRNA1.1 into small-vault RNAs (svRNAs). We identify the serine/arginine rich splicing factor 2 (SRSF2) as a novel VTRNA1.1-binding protein that counteracts VTRNA1.1 processing by binding the non-methylated form with higher affinity. Both NSUN2 and SRSF2 orchestrate the production of distinct svRNAs. Finally, we discover a functional role of svRNAs in regulating the epidermal differentiation programme. Thus, our data reveal a direct role for m5C in the processing of VTRNA1.1 that involves SRSF2 and is crucial for efficient cellular differentiation.
    DOI:  https://doi.org/10.1038/s41467-019-10020-7
  48. Cell Res. 2019 Jun 12.
    Mitochondrial metabolism and glutamine are essential for mesoderm differentiation of human pluripotent stem cells.
    Vivian Lu, Perrine Dahan, Fasih M Ahsan, Alexander N Patananan, Irena J Roy, Alejandro Torres, Robert M T Nguyen, Dian Huang, Daniel Braas, Michael A Teitell.
      
    DOI:  https://doi.org/10.1038/s41422-019-0191-2
  49. Sci Signal. 2019 Jun 11. pii: eaav3249. [Epub ahead of print]12(585):
    AMPK directly activates mTORC2 to promote cell survival during acute energetic stress.
    Dubek Kazyken, Brian Magnuson, Cagri Bodur, Hugo A Acosta-Jaquez, Deqiang Zhang, Xin Tong, Tammy M Barnes, Gabrielle K Steinl, Nicole E Patterson, Christopher H Altheim, Naveen Sharma, Ken Inoki, Gregory D Cartee, Dave Bridges, Lei Yin, Steven M Riddle, Diane C Fingar.
      AMP-activated protein kinase (AMPK) senses energetic stress and, in turn, promotes catabolic and suppresses anabolic metabolism coordinately to restore energy balance. We found that a diverse array of AMPK activators increased mTOR complex 2 (mTORC2) signaling in an AMPK-dependent manner in cultured cells. Activation of AMPK with the type 2 diabetes drug metformin (GlucoPhage) also increased mTORC2 signaling in liver in vivo and in primary hepatocytes in an AMPK-dependent manner. AMPK-mediated activation of mTORC2 did not result from AMPK-mediated suppression of mTORC1 and thus reduced negative feedback on PI3K flux. Rather, AMPK associated with and directly phosphorylated mTORC2 (mTOR in complex with rictor). As determined by two-stage in vitro kinase assay, phosphorylation of mTORC2 by recombinant AMPK was sufficient to increase mTORC2 catalytic activity toward Akt. Hence, AMPK phosphorylated mTORC2 components directly to increase mTORC2 activity and downstream signaling. Functionally, inactivation of AMPK, mTORC2, and Akt increased apoptosis during acute energetic stress. By showing that AMPK activates mTORC2 to increase cell survival, these data provide a potential mechanism for how AMPK paradoxically promotes tumorigenesis in certain contexts despite its tumor-suppressive function through inhibition of growth-promoting mTORC1. Collectively, these data unveil mTORC2 as a target of AMPK and the AMPK-mTORC2 axis as a promoter of cell survival during energetic stress.
    DOI:  https://doi.org/10.1126/scisignal.aav3249
  50. BMB Rep. 2019 Jun 12. pii: 4657. [Epub ahead of print]
    Metabolic influence on macrophage polarization and pathogenesis.
    Bikash Thapa, Keunwook Lee.
      Macrophages play an essential role not only in mediating the first line of defense but also in maintaining tissue homeostasis. In response to extrinsic factors derived from a given tissue, macrophages activate different functional programs to produce polarized macrophage populations responsible for inducing inflammation against microbes, removing cellular debris, and tissue repair. However, accumulating evidence has revealed that macrophage polarization is pivotal in the pathophysiology of metabolic syndromes and cancer, as well as in infectious and autoimmune diseases. Recent advances in transcriptomic and metabolomic studies have highlighted the link between metabolic rewiring of macrophages and their functional plasticity. These findings imply that metabolic adaption to their surrounding microenvironment instructs activation of macrophages with functionally distinct phenotypes, which in turn probably leads to the pathogenesis of a wide spectrum of diseases. In this review, we have introduced emerging concepts in immunometabolism with focus on the impact on functional activation of macrophages. Furthermore, we have discussed the implication of macrophage plasticity on the pathogenesis of metabolic syndromes and cancer, and how the disease microenvironment manipulates macrophage metabolism with regard to the pathophysiology.
This page is being maintained by Thomas Krichel. It was last updated on 2024‒11‒22 at 4:13.