bims-camemi 2019-08-04 papers

bims-camemi

Biomed News

on Mitochondrial metabolism in cancer

Issue of 2019–08–04
thirty-six papers selected by
Christian Frezza, , University of Cambridge, MRC Cancer Unit

Dietary methionine influences therapy in mouse cancer models and alters human metabolism.
Dynamic regulation of subcellular mitochondrial position for localized metabolite levels.
Resolving subcellular plant metabolism.
Selective Autophagy of Mitochondria on a Ubiquitin-Endoplasmic-Reticulum Platform.
Activation of PINK1-Parkin-Mediated Mitophagy Degrades Mitochondrial Quality Control Proteins in Fuchs Endothelial Corneal Dystrophy.
Connecting lysosomes and mitochondria - a novel role for lipid metabolism in cancer cell death.
Role of p110a subunit of PI3-kinase in skeletal muscle mitochondrial homeostasis and metabolism.
IMP dehydrogenase-2 drives aberrant nucleolar activity and promotes tumorigenesis in glioblastoma.
TREM1/3 Deficiency Impairs Tissue Repair After Acute Kidney Injury and Mitochondrial Metabolic Flexibility in Tubular Epithelial Cells.
ERp18 regulates activation of ATF6α during unfolded protein response.
MITOL prevents ER stress-induced apoptosis by IRE1α ubiquitylation at ER-mitochondria contact sites.
Lysine harvesting is an antioxidant strategy and triggers underground polyamine metabolism.
Clockophagy is a novel selective autophagy process favoring ferroptosis.
Serine-Glycine-One-Carbon Metabolism: The Hidden Achilles Heel of MYCN-Amplified Neuroblastoma?
mTORC1-PGC1 axis regulates mitochondrial remodeling during reprogramming.
Metabolic Reprogramming Promotes Myogenesis During Aging.
Loss of p53 triggers WNT-dependent systemic inflammation to drive breast cancer metastasis.
Dynamic polyhedral actomyosin lattices remodel micron-scale curved membranes during exocytosis in live mice.
Modulation of complex II-energized respiration in muscle, heart, and brown adipose mitochondria by oxaloacetate and complex I electron flow.
NDUFAB1 confers cardio-protection by enhancing mitochondrial bioenergetics through coordination of respiratory complex and supercomplex assembly.
Linking Cancer Metabolic Dysfunction and Genetic Instability through the Lens of Iron Metabolism.
Should we consider subcellular compartmentalization of metabolites, and if so, how do we measure them?
Quantitative Profiling of Oncometabolites in Frozen and Formalin-Fixed Paraffin-Embedded Tissue Specimens by Liquid Chromatography Coupled with Tandem Mass Spectrometry.
5-methylcytosine promotes pathogenesis of bladder cancer through stabilizing mRNAs.
Targeting the Mitochondrial Potassium Channel Kv1.3 to Kill Cancer Cells: Drugs, Strategies, and New Perspectives.
Mitofusins regulate lipid metabolism to mediate the development of lung fibrosis.
Mitochondrial functionality and metabolism in T cells from progressive multiple sclerosis patients.
MYC paralog-dependent apoptotic priming orchestrates a spectrum of vulnerabilities in small cell lung cancer.
Hypothalamic mTORC2 is essential for metabolic health and longevity.
Glutamine Synthetase Promotes Radiation Resistance via Facilitating Nucleotide Metabolism and Subsequent DNA Damage Repair.
Targeting Pathogenic Lafora Bodies in Lafora Disease Using an Antibody-Enzyme Fusion.
Repressive Gene Regulation Synchronizes Development with Cellular Metabolism.
Defects of mitochondrial RNA turnover lead to the accumulation of double-stranded RNA in vivo.
The Circadian Protein Nocturnin Regulates Metabolic Adaptation in Brown Adipose Tissue.
De-fusing mitochondria defuses the mtDNA time-bomb.
Identification and Application of Gene Expression Signatures Associated with Lifespan Extension.

Nature. 2019 Jul 31.

Dietary methionine influences therapy in mouse cancer models and alters human metabolism.

Xia Gao, Sydney M Sanderson, Ziwei Dai, Michael A Reid, Daniel E Cooper, Min Lu, John P Richie, Amy Ciccarella, Ana Calcagnotto, Peter G Mikhael, Samantha J Mentch, Juan Liu, Gene Ables, David G Kirsch, David S Hsu, Sailendra N Nichenametla, Jason W Locasale.

Nutrition exerts considerable effects on health, and dietary interventions are commonly used to treat diseases of metabolic aetiology. Although cancer has a substantial metabolic component1, the principles that define whether nutrition may be used to influence outcomes of cancer are unclear2. Nevertheless, it is established that targeting metabolic pathways with pharmacological agents or radiation can sometimes lead to controlled therapeutic outcomes. By contrast, whether specific dietary interventions can influence the metabolic pathways that are targeted in standard cancer therapies is not known. Here we show that dietary restriction of the essential amino acid methionine-the reduction of which has anti-ageing and anti-obesogenic properties-influences cancer outcome, through controlled and reproducible changes to one-carbon metabolism. This pathway metabolizes methionine and is the target of a variety of cancer interventions that involve chemotherapy and radiation. Methionine restriction produced therapeutic responses in two patient-derived xenograft models of chemotherapy-resistant RAS-driven colorectal cancer, and in a mouse model of autochthonous soft-tissue sarcoma driven by a G12D mutation in KRAS and knockout of p53 (KrasG12D/+;Trp53-/-) that is resistant to radiation. Metabolomics revealed that the therapeutic mechanisms operate via tumour-cell-autonomous effects on flux through one-carbon metabolism that affects redox and nucleotide metabolism-and thus interact with the antimetabolite or radiation intervention. In a controlled and tolerated feeding study in humans, methionine restriction resulted in effects on systemic metabolism that were similar to those obtained in mice. These findings provide evidence that a targeted dietary manipulation can specifically affect tumour-cell metabolism to mediate broad aspects of cancer outcome.

DOI: https://doi.org/10.1038/s41586-019-1437-3
J Biochem. 2019 Jul 29. pii: mvz058. [Epub ahead of print]

Dynamic regulation of subcellular mitochondrial position for localized metabolite levels.

Haya Alshaabi, Meara Heininger, Brian Cunniff.

  Mitochondria are not passive bystanders aimlessly floating throughout our cell's cytoplasm. Instead, mitochondria actively move, anchor, divide, fuse, self-destruct and transfer between cells in a coordinated fashion, all to ensure proper structure and position supporting cell function. The existence of the mitochondria in our cells has long been appreciated, but their dynamic nature and interaction with other subcellular compartments has only recently been fully realized with the advancement of high resolution live-cell microscopy and improved fractionization techniques. The how and why that dictates positioning of mitochondria to specific subcellular sites is an ever-expanding research area. Furthermore, the advent of new and improved functional probes, sensitive to changes in subcellular metabolite levels has increased our understanding of local mitochondrial populations. In this review we will address the evidence for intentional mitochondrial positioning in supporting subcellular mitochondrial metabolite levels, including calcium (Ca2+), adenosine triphosphate (ATP) and reactive oxygen species (ROS) and the role they play in dictating cell outcomes.

Keywords:  Mitochondrial dynamics; metabolite gradients; mitochondrial contacts; reactive oxygen species

DOI:  https://doi.org/10.1093/jb/mvz058
Plant J. 2019 Jul 30.

Resolving subcellular plant metabolism.

Lisa Fürtauer, Lisa Küstner, Wolfram Weckwerth, Arnd G Heyer, Thomas Nägele.

  Plant cells are characterized by a high degree of compartmentalization and a diverse proteome and metabolome. Only a very limited number of studies has addressed combined subcellular proteomics and metabolomics which strongly limits biochemical and physiological interpretation of large-scale omics data. Our study presents a methodological combination of non-aqueous fractionation, shotgun proteomics, enzyme activities and metabolomics to reveal subcellular diurnal dynamics of plant metabolism. Subcellular marker protein sets were identified and enzymatically validated to resolve metabolism in a 4-compartment model comprising chloroplasts, cytosol, vacuole and mitochondria. These marker sets are now available for future studies which aim to monitor subcellular metabolome and proteome dynamics. Comparing subcellular dynamics in wild type plants and HXK1-deficient gin2-1 mutants revealed a strong impact of HXK1 activity on metabolome dynamics in multiple compartments. Glucose accumulation in the cytosol of gin2-1 was accompanied by diminished vacuolar glucose levels. Subcellular dynamics of pyruvate, succinate and fumarate amount was significantly affected in gin2-1 which coincided with differential mitochondrial proteome dynamics. Lowered mitochondrial glycine and serine amount in gin2-1 together with reduced abundance of photorespiratory proteins indicated an effect of the gin2-1 mutation on photorespiratory capacity. Our findings highlight the necessity to resolve plant metabolism to a subcellular level in order to provide a causal relationship between metabolites, proteins and metabolic pathway regulation. This article is protected by copyright. All rights reserved.

Keywords:   Arabidopsis thaliana ; hexokinase 1; metabolomics; non-aqueous fractionation; photorespiration; proteomics; subcellular metabolism

DOI:  https://doi.org/10.1111/tpj.14472
Dev Cell. 2019 Jul 15. pii: S1534-5807(19)30569-6. [Epub ahead of print]

Selective Autophagy of Mitochondria on a Ubiquitin-Endoplasmic-Reticulum Platform.

Maria Zachari, Sigurdur R Gudmundsson, Ziyue Li, Maria Manifava, Ronak Shah, Matthew Smith, James Stronge, Eleftherios Karanasios, Caterina Piunti, Chieko Kishi-Itakura, Helena Vihinen, Eija Jokitalo, Jun-Lin Guan, Folma Buss, Andrew M Smith, Simon A Walker, Eeva-Liisa Eskelinen, Nicholas T Ktistakis.

  The dynamics and coordination between autophagy machinery and selective receptors during mitophagy are unknown. Also unknown is whether mitophagy depends on pre-existing membranes or is triggered on the surface of damaged mitochondria. Using a ubiquitin-dependent mitophagy inducer, the lactone ivermectin, we have combined genetic and imaging experiments to address these questions. Ubiquitination of mitochondrial fragments is required the earliest, followed by auto-phosphorylation of TBK1. Next, early essential autophagy proteins FIP200 and ATG13 act at different steps, whereas ULK1 and ULK2 are dispensable. Receptors act temporally and mechanistically upstream of ATG13 but downstream of FIP200. The VPS34 complex functions at the omegasome step. ATG13 and optineurin target mitochondria in a discontinuous oscillatory way, suggesting multiple initiation events. Targeted ubiquitinated mitochondria are cradled by endoplasmic reticulum (ER) strands even without functional autophagy machinery and mitophagy adaptors. We propose that damaged mitochondria are ubiquitinated and dynamically encased in ER strands, providing platforms for formation of the mitophagosomes.

Keywords:  autophagosome; autophagy; endoplasmic reticulum; mitophagy; super resolution microscopy; tomography

DOI:  https://doi.org/10.1016/j.devcel.2019.06.016
Am J Pathol. 2019 Jul 27. pii: S0002-9440(19)30653-4. [Epub ahead of print]

Activation of PINK1-Parkin-Mediated Mitophagy Degrades Mitochondrial Quality Control Proteins in Fuchs Endothelial Corneal Dystrophy.

Takashi Miyai, Shivakumar Vasanth, Geetha Melangath, Neha Deshpande, Varun Kumar, Anne-Sophie Benischke, Yuming Chen, Marianne O Price, Francis W Price, Ula V Jurkunas.

Corneal endothelium (CE) is a monolayer of mitochondria-rich cells, critical for maintaining corneal transparency compatible with clear vision. Fuchs endothelial corneal dystrophy (FECD) is a heterogeneous genetically complex disorder, where oxidative stress plays a key role in the rosette formation during the degenerative loss of CE. Increased mitochondrial fragmentation along with excessive mitophagy activation has been detected in FECD; however, the mechanism of aberrant mitochondrial dynamics in CE cell loss is poorly understood. Here, we investigate the role of oxidative stress in mitophagy activation in FECD. Immunoblotting of FECD ex vivo specimens revealed an accumulation of PINK1 and phospho-Parkin (Ser65) along with loss of total Parkin and total Drp1. Similarly, modeling of rosette formation with menadione (MN), led to phospho-Parkin accumulation in fragmented mitochondria resulting in mitophagy-induced mitochondrial clearance, albeit possibly in a PINK1 independent manner. Loss of PINK1, phospho-Drp1 and total Drp1 was prominent after MN-induced oxidative stress, but not after mitochondrial depolarization by carbonyl cyanide m-chlorophenyl hydrazone (CCCP). Moreover, MN-induced mitophagy led to degradation of Parkin along with sequestration of Drp1 and PINK1 that was rescued by mitophagy inhibition. This study shows that in FECD intracellular oxidative stress induces Parkin-mediated mitochondrial fragmentation where endogenous Drp1 and PINK1 are sequestered and degraded by mitophagy during degenerative loss of post-mitotic cells of ocular tissue.

DOI: https://doi.org/10.1016/j.ajpath.2019.06.012
Cell Commun Signal. 2019 Jul 29. 17(1): 87

Connecting lysosomes and mitochondria - a novel role for lipid metabolism in cancer cell death.

Karin Bartel, Helmut Pein, Bastian Popper, Sabine Schmitt, Sudha Janaki-Raman, Almut Schulze, Florian Lengauer, Andreas Koeberle, Oliver Werz, Hans Zischka, Rolf Müller, Angelika M Vollmar, Karin von Schwarzenberg.

   BACKGROUND: The understanding of lysosomes has been expanded in recent research way beyond their view as cellular trash can. Lysosomes are pivotal in regulating metabolism, endocytosis and autophagy and are implicated in cancer. Recently it was discovered that the lysosomal V-ATPase, which is known to induce apoptosis, interferes with lipid metabolism in cancer, yet the interplay between these organelles is poorly understood.
METHODS: LC-MS/MS analysis was performed to investigate lipid distribution in cells. Cell survival and signaling pathways were analyzed by means of cell biological methods (qPCR, Western Blot, flow cytometry, CellTiter-Blue). Mitochondrial structure was analyzed by confocal imaging and electron microscopy, their function was determined by flow cytometry and seahorse measurements.
RESULTS: Our data reveal that interfering with lysosomal function changes composition and subcellular localization of triacylglycerids accompanied by an upregulation of PGC1α and PPARα expression, master regulators of energy and lipid metabolism. Furthermore, cardiolipin content is reduced driving mitochondria into fission, accompanied by a loss of membrane potential and reduction in oxidative capacity, which leads to a deregulation in cellular ROS and induction of mitochondria-driven apoptosis. Additionally, cells undergo a metabolic shift to glutamine dependency, correlated with the fission phenotype and sensitivity to lysosomal inhibition, most prominent in Ras mutated cells.
CONCLUSION: This study sheds mechanistic light on a largely uninvestigated triangle between lysosomes, lipid metabolism and mitochondrial function. Insight into this organelle crosstalk increases our understanding of mitochondria-driven cell death. Our findings furthermore provide a first hint on a connection of Ras pathway mutations and sensitivity towards lysosomal inhibitors.

Keywords:  Apoptosis; Cardiolipin; Fission; Lipid metabolism; Lysosome; Mitochondria; V-ATPase

DOI:  https://doi.org/10.1186/s12964-019-0399-2
Nat Commun. 2019 Jul 30. 10(1): 3412

Role of p110a subunit of PI3-kinase in skeletal muscle mitochondrial homeostasis and metabolism.

Mengyao Ella Li, Hans P M M Lauritzen, Brian T O'Neill, Chih-Hao Wang, Weikang Cai, Bruna B Brandao, Masaji Sakaguchi, Rongya Tao, Michael F Hirshman, Samir Softic, C Ronald Kahn.

Skeletal muscle insulin resistance, decreased phosphatidylinositol 3-kinase (PI3K) activation and altered mitochondrial function are hallmarks of type 2 diabetes. To determine the relationship between these abnormalities, we created mice with muscle-specific knockout of the p110α or p110β catalytic subunits of PI3K. We find that mice with muscle-specific knockout of p110α, but not p110β, display impaired insulin signaling and reduced muscle size due to enhanced proteasomal and autophagic activity. Despite insulin resistance and muscle atrophy, M-p110αKO mice show decreased serum myostatin, increased mitochondrial mass, increased mitochondrial fusion, and increased PGC1α expression, especially PCG1α2 and PCG1α3. This leads to enhanced mitochondrial oxidative capacity, increased muscle NADH content, and higher muscle free radical release measured in vivo using pMitoTimer reporter. Thus, p110α is the dominant catalytic isoform of PI3K in muscle in control of insulin sensitivity and muscle mass, and has a unique role in mitochondrial homeostasis in skeletal muscle.

DOI: https://doi.org/10.1038/s41467-019-11265-y
Nat Cell Biol. 2019 Aug;21(8): 1003-1014

IMP dehydrogenase-2 drives aberrant nucleolar activity and promotes tumorigenesis in glioblastoma.

Satoshi Kofuji, Akiyoshi Hirayama, Alexander Otto Eberhardt, Risa Kawaguchi, Yuki Sugiura, Oltea Sampetrean, Yoshiki Ikeda, Mikako Warren, Naoya Sakamoto, Shuji Kitahara, Hirofumi Yoshino, Daisuke Yamashita, Kazutaka Sumita, Kara Wolfe, Lisa Lange, Satsuki Ikeda, Hiroko Shimada, Noriaki Minami, Akshiv Malhotra, Shin Morioka, Yuki Ban, Maya Asano, Victoria L Flanary, Annmarie Ramkissoon, Lionel M L Chow, Juri Kiyokawa, Tomoyuki Mashimo, Greg Lucey, Sergey Mareninov, Tatsuya Ozawa, Nobuyuki Onishi, Koichi Okumura, Jumpei Terakawa, Takiko Daikoku, Trisha Wise-Draper, Nazanin Majd, Kaori Kofuji, Mika Sasaki, Masaru Mori, Yonehiro Kanemura, Eric P Smith, Dimitrios Anastasiou, Hiroaki Wakimoto, Eric C Holland, William H Yong, Craig Horbinski, Ichiro Nakano, Ralph J DeBerardinis, Robert M Bachoo, Paul S Mischel, Wataru Yasui, Makoto Suematsu, Hideyuki Saya, Tomoyoshi Soga, Ingrid Grummt, Holger Bierhoff, Atsuo T Sasaki.

In many cancers, high proliferation rates correlate with elevation of rRNA and tRNA levels, and nucleolar hypertrophy. However, the underlying mechanisms linking increased nucleolar transcription and tumorigenesis are only minimally understood. Here we show that IMP dehydrogenase-2 (IMPDH2), the rate-limiting enzyme for de novo guanine nucleotide biosynthesis, is overexpressed in the highly lethal brain cancer glioblastoma. This leads to increased rRNA and tRNA synthesis, stabilization of the nucleolar GTP-binding protein nucleostemin, and enlarged, malformed nucleoli. Pharmacological or genetic inactivation of IMPDH2 in glioblastoma reverses these effects and inhibits cell proliferation, whereas untransformed glia cells are unaffected by similar IMPDH2 perturbations. Impairment of IMPDH2 activity triggers nucleolar stress and growth arrest of glioblastoma cells even in the absence of functional p53. Our results reveal that upregulation of IMPDH2 is a prerequisite for the occurance of aberrant nucleolar function and increased anabolic processes in glioblastoma, which constitutes a primary event in gliomagenesis.

DOI: https://doi.org/10.1038/s41556-019-0363-9
Front Immunol. 2019 ;10 1469

TREM1/3 Deficiency Impairs Tissue Repair After Acute Kidney Injury and Mitochondrial Metabolic Flexibility in Tubular Epithelial Cells.

Alessandra Tammaro, Angelique M L Scantlebery, Elena Rampanelli, Cristiana Borrelli, Nike Claessen, Loes M Butter, Alessandra Soriani, Marco Colonna, Jaklien C Leemans, Mark C Dessing, Sandrine Florquin.

  Long-term sequelae of acute kidney injury (AKI) are associated with incomplete recovery of renal function and the development of chronic kidney disease (CKD), which can be mediated by aberrant innate immune activation, mitochondrial pathology, and accumulation of senescent tubular epithelial cells (TECs). Herein, we show that the innate immune receptor Triggering receptor expressed on myeloid cells-1 (TREM-1) links mitochondrial metabolism to tubular epithelial senescence. TREM-1 is expressed by inflammatory and epithelial cells, both players in renal repair after ischemia/reperfusion (IR)-induced AKI. Hence, we subjected WT and TREM1/3 KO mice to different models of renal IR. TREM1/3 KO mice displayed no major differences during the acute phase of injury, but increased mortality was observed in the recovery phase. This detrimental effect was associated with maladaptive repair, characterized by persistent tubular damage, inflammation, fibrosis, and TEC senescence. In vitro, we observed an altered mitochondrial homeostasis and cellular metabolism in TREM1/3 KO primary TECs. This was associated with G2/M arrest and increased ROS accumulation. Further exposure of cells to ROS-generating triggers drove the cells into a stress-induced senescent state, resulting in decreased wound healing capacity. Treatment with a mitochondria anti-oxidant partly prevented the senescent phenotype, suggesting a role for mitochondria herein. In summary, we have unraveled a novel (metabolic) mechanism by which TREM1/3 deficiency drives senescence in TECs. This involves redox imbalance, mitochondrial dysfunction and a decline in cellular metabolic activities. These finding suggest a novel role for TREM-1 in maintaining tubular homeostasis through regulation of mitochondrial metabolic flexibility.

Keywords:  epithelial innate immunity; ischemia/reperfusion injury; maladaptive repair; mitochondrial metabolism; renal repair; tubular cell senescence

DOI:  https://doi.org/10.3389/fimmu.2019.01469
EMBO J. 2019 Aug 01. 38(15): e100990

ERp18 regulates activation of ATF6α during unfolded protein response.

Ojore Bv Oka, Marcel van Lith, Jana Rudolf, Wanida Tungkum, Marie Anne Pringle, Neil J Bulleid.

  Activation of the ATF6α signaling pathway is initiated by trafficking of ATF6α from the ER to the Golgi apparatus. Its subsequent proteolysis releases a transcription factor that translocates to the nucleus causing downstream gene activation. How ER retention, Golgi trafficking, and proteolysis of ATF6α are regulated and whether additional protein partners are required for its localization and processing remain unresolved. Here, we show that ER-resident oxidoreductase ERp18 associates with ATF6α following ER stress and plays a key role in both trafficking and activation of ATF6α. We find that ERp18 depletion attenuates the ATF6α stress response. Paradoxically, ER stress accelerates trafficking of ATF6α to the Golgi in ERp18-depleted cells. However, the translocated ATF6α becomes aberrantly processed preventing release of the soluble transcription factor. Hence, we demonstrate that ERp18 monitors ATF6α ER quality control to ensure optimal processing following trafficking to the Golgi.

Keywords:  ATF6α; ER stress; ERp18; protein trafficking; unfolded protein response

DOI:  https://doi.org/10.15252/embj.2018100990
EMBO J. 2019 Aug 01. 38(15): e100999

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; mitochondria-associated ER membrane; mitochondrial E3 ligase MITOL/MARCH5; unfolded protein response

DOI:  https://doi.org/10.15252/embj.2018100999
Nature. 2019 Jul 31.

Lysine harvesting is an antioxidant strategy and triggers underground polyamine metabolism.

Viridiana Olin-Sandoval, Jason Shu Lim Yu, Leonor Miller-Fleming, Mohammad Tauqeer Alam, Stephan Kamrad, Clara Correia-Melo, Robert Haas, Joanna Segal, David Alejandro Peña Navarro, Lucia Herrera-Dominguez, Oscar Méndez-Lucio, Jakob Vowinckel, Michael Mülleder, Markus Ralser.

Both single and multicellular organisms depend on anti-stress mechanisms that enable them to deal with sudden changes in the environment, including exposure to heat and oxidants. Central to the stress response are dynamic changes in metabolism, such as the transition from the glycolysis to the pentose phosphate pathway-a conserved first-line response to oxidative insults1,2. Here we report a second metabolic adaptation that protects microbial cells in stress situations. The role of the yeast polyamine transporter Tpo1p3-5 in maintaining oxidant resistance is unknown6. However, a proteomic time-course experiment suggests a link to lysine metabolism. We reveal a connection between polyamine and lysine metabolism during stress situations, in the form of a promiscuous enzymatic reaction in which the first enzyme of the polyamine pathway, Spe1p, decarboxylates lysine and forms an alternative polyamine, cadaverine. The reaction proceeds in the presence of extracellular lysine, which is taken up by cells to reach concentrations up to one hundred times higher than those required for growth. Such extensive harvest is not observed for the other amino acids, is dependent on the polyamine pathway and triggers a reprogramming of redox metabolism. As a result, NADPH-which would otherwise be required for lysine biosynthesis-is channelled into glutathione metabolism, leading to a large increase in glutathione concentrations, lower levels of reactive oxygen species and increased oxidant tolerance. Our results show that nutrient uptake occurs not only to enable cell growth, but when the nutrient availability is favourable it also enables cells to reconfigure their metabolism to preventatively mount stress protection.

DOI: https://doi.org/10.1038/s41586-019-1442-6
Sci Adv. 2019 Jul;5(7): eaaw2238

Clockophagy is a novel selective autophagy process favoring ferroptosis.

Minghua Yang, Pan Chen, Jiao Liu, Shan Zhu, Guido Kroemer, Daniel J Klionsky, Michael T Lotze, Herbert J Zeh, Rui Kang, Daolin Tang.

Ferroptosis is a form of nonapoptotic regulated cell death driven by iron-dependent lipid peroxidation. Autophagy involves a lysosomal degradation pathway that can either promote or impede cell death. A high level of autophagy has been associated with ferroptosis, but the mechanisms underpinning this relationship are largely elusive. We characterize the contribution of autophagy to ferroptosis in human cancer cell lines and mouse tumor models. We show that "clockophagy," the selective degradation of the core circadian clock protein ARNTL by autophagy, is critical for ferroptosis. We identify SQSTM1 as a cargo receptor responsible for autophagic ARNTL degradation. ARNTL inhibits ferroptosis by repressing the transcription of Egln2, thus activating the prosurvival transcription factor HIF1A. Genetic or pharmacological interventions blocking ARNTL degradation or inhibiting EGLN2 activation diminished, whereas destabilizing HIF1A facilitated, ferroptotic tumor cell death. Thus, our findings reveal a new pathway, initiated by the autophagic removal of ARNTL, that facilitates ferroptosis induction.

DOI: https://doi.org/10.1126/sciadv.aaw2238
Cancer Res. 2019 Aug 01. 79(15): 3818-3819

Serine-Glycine-One-Carbon Metabolism: The Hidden Achilles Heel of MYCN-Amplified Neuroblastoma?

Aida Rodriguez Garcia, Marie Arsenian-Henriksson.

In this issue of Cancer Research, Xia and colleagues show that MYC-induced metabolic reprograming results in dependency on the serine-glycine-one-carbon (SGOC) metabolic pathway in neuroblastoma. This occurs through MYCN and ATF4 activation of the SGOC biosynthetic pathway in MYCN-amplified cells. Furthermore, inhibition of de novo serine synthesis generates metabolic stress in MYCN-amplified neuroblastoma cells, causing cell-cycle arrest and autophagy. Together, these data suggest that the SGOC pathway is an attractive therapy target in neuroblastoma.See related article by Xia et al., p. 3837.

DOI: https://doi.org/10.1158/0008-5472.CAN-19-1816
FEBS J. 2019 Jul 30.

mTORC1-PGC1 axis regulates mitochondrial remodeling during reprogramming.

Lu Lu Wang, Xue Ting Xu, Che Jiang, Gang Ma, Ying Hua Huang, Hui Zhang, Yi Wei Lai, Ming Wang, Tanveer Ahmed, Run Xia Lin, Wen Jing Guo, Zhi Wei Luo, Wen Juan Li, Meng Zhang, Carl Ward, Min Xian Qian, Baohua Liu, Miguel A Esteban, Baoming Qin.

  Metabolic reprogramming, hallmarked by enhanced glycolysis and reduced mitochondrial activity, is a key event in the early phase of somatic cell reprogramming. Although extensive work has been conducted to identify the mechanisms of mitochondrial remodeling in reprogramming, many questions remain. In this regard, different laboratories have proposed a role in this process for either canonical (ATG5-dependent) autophagy-mediated mitochondrial degradation (mitophagy), non-canonical (ULK1-dependent, ATG5-independent) mitophagy, mitochondrial fission or reduced biogenesis due to mTORC1 suppression. Clarifying these discrepancies is important for providing a comprehensive picture of metabolic changes in reprogramming. Yet, the comparison among these studies is difficult because they use different reprogramming conditions and mitophagy detection/quantification methods. Here, we have systematically explored mitochondrial remodeling in reprogramming using different culture media and reprogramming factor cocktails, together with appropriate quantification methods and thorough statistical analysis. Our experiments show lack of evidence for mitophagy in mitochondrial remodeling in reprogramming, and further confirm that the suppression of the mTORC1-PGC1 pathway drives this process. Our work helps to clarify the complex interplay between metabolic changes and nutrient sensing pathways in reprogramming, which may also shed light on other contexts such as development, aging and cancer. This article is protected by copyright. All rights reserved.

Keywords:  PGC1; Reprogramming; mTORC1; mitochondrial remodeling; mitophagy

DOI:  https://doi.org/10.1111/febs.15024
Front Physiol. 2019 ;10 897

Metabolic Reprogramming Promotes Myogenesis During Aging.

Roberta Belli, Agnese Bonato, Luciana De Angelis, Simone Mirabilii, Maria Rosaria Ricciardi, Agostino Tafuri, Alessio Molfino, Massimiliano Leigheb, Paola Costelli, Maurizia Caruso, Maurizio Muscaritoli, Elisabetta Ferraro.

  Sarcopenia is the age-related progressive loss of skeletal muscle mass and strength finally leading to poor physical performance. Impaired myogenesis contributes to the pathogenesis of sarcopenia, while mitochondrial dysfunctions are thought to play a primary role in skeletal muscle loss during aging. Here we studied the link between myogenesis and metabolism. In particular, we analyzed the effect of the metabolic modulator trimetazidine (TMZ) on myogenesis in aging. We show that reprogramming the metabolism by TMZ treatment for 12 consecutive days stimulates myogenic gene expression in skeletal muscle of 22-month-old mice. Our data also reveal that TMZ increases the levels of mitochondrial proteins and stimulates the oxidative metabolism in aged muscles, this finding being in line with our previous observations in cachectic mice. Moreover, we show that, besides TMZ also other types of metabolic modulators (i.e., 5-Aminoimidazole-4-Carboxamide Ribofuranoside-AICAR) can stimulate differentiation of skeletal muscle progenitors in vitro. Overall, our results reveal that reprogramming the metabolism stimulates myogenesis while triggering mitochondrial proteins synthesis in vivo during aging. Together with the previously reported ability of TMZ to increase muscle strength in aged mice, these new data suggest an interesting non-invasive therapeutic strategy which could contribute to improving muscle quality and neuromuscular communication in the elderly, and counteracting sarcopenia.

Keywords:  aging; metabolic reprogramming; metabolism; mitochondria; myogenesis; neuromuscular activity; sarcopenia; trimetazidine

DOI:  https://doi.org/10.3389/fphys.2019.00897
Nature. 2019 Jul 31.

Loss of p53 triggers WNT-dependent systemic inflammation to drive breast cancer metastasis.

Max D Wellenstein, Seth B Coffelt, Danique E M Duits, Martine H van Miltenburg, Maarten Slagter, Iris de Rink, Linda Henneman, Sjors M Kas, Stefan Prekovic, Cheei-Sing Hau, Kim Vrijland, Anne Paulien Drenth, Renske de Korte-Grimmerink, Eva Schut, Ingrid van der Heijden, Wilbert Zwart, Lodewyk F A Wessels, Ton N Schumacher, Jos Jonkers, Karin E de Visser.

Cancer-associated systemic inflammation is strongly linked to poor disease outcome in patients with cancer1,2. For most human epithelial tumour types, high systemic neutrophil-to-lymphocyte ratios are associated with poor overall survival3, and experimental studies have demonstrated a causal relationship between neutrophils and metastasis4,5. However, the cancer-cell-intrinsic mechanisms that dictate the substantial heterogeneity in systemic neutrophilic inflammation between tumour-bearing hosts are largely unresolved. Here, using a panel of 16 distinct genetically engineered mouse models for breast cancer, we uncover a role for cancer-cell-intrinsic p53 as a key regulator of pro-metastatic neutrophils. Mechanistically, loss of p53 in cancer cells induced the secretion of WNT ligands that stimulate tumour-associated macrophages to produce IL-1β, thus driving systemic inflammation. Pharmacological and genetic blockade of WNT secretion in p53-null cancer cells reverses macrophage production of IL-1β and subsequent neutrophilic inflammation, resulting in reduced metastasis formation. Collectively, we demonstrate a mechanistic link between the loss of p53 in cancer cells, secretion of WNT ligands and systemic neutrophilia that potentiates metastatic progression. These insights illustrate the importance of the genetic makeup of breast tumours in dictating pro-metastatic systemic inflammation, and set the stage for personalized immune intervention strategies for patients with cancer.

DOI: https://doi.org/10.1038/s41586-019-1450-6
Nat Cell Biol. 2019 Aug;21(8): 933-939

Dynamic polyhedral actomyosin lattices remodel micron-scale curved membranes during exocytosis in live mice.

Seham Ebrahim, Desu Chen, Max Weiss, Lenka Malec, Yeap Ng, Ivan Rebustini, Evan Krystofiak, Longhua Hu, Jian Liu, Andrius Masedunskas, Edna Hardeman, Peter Gunning, Bechara Kachar, Roberto Weigert.

Actomyosin networks, the cell's major force production machineries, remodel cellular membranes during myriad dynamic processes1,2 by assembling into various architectures with distinct force generation properties3,4. While linear and branched actomyosin architectures are well characterized in cell-culture and cell-free systems3, it is not known how actin and myosin networks form and function to remodel membranes in complex three-dimensional mammalian tissues. Here, we use four-dimensional spinning-disc confocal microscopy with image deconvolution to acquire macromolecular-scale detail of dynamic actomyosin networks in exocrine glands of live mice. We address how actin and myosin organize around large membrane-bound secretory vesicles and generate the forces required to complete exocytosis5-7. We find that actin and non-muscle myosin II (NMII) assemble into previously undescribed polyhedral-like lattices around the vesicle membrane. The NMII lattice comprises bipolar minifilaments8-10 as well as non-canonical three-legged configurations. Using photobleaching and pharmacological perturbations in vivo, we show that actomyosin contractility and actin polymerization together push on the underlying vesicle membrane to overcome the energy barrier and complete exocytosis7. Our imaging approach thus unveils a force-generating actomyosin lattice that regulates secretion in the exocrine organs of live animals.

DOI: https://doi.org/10.1038/s41556-019-0365-7
FASEB J. 2019 Aug 01. fj201900690R

Modulation of complex II-energized respiration in muscle, heart, and brown adipose mitochondria by oxaloacetate and complex I electron flow.

Brian D Fink, Liping Yu, William I Sivitz.

  We recently reported that membrane potential (ΔΨ) primarily determines the relationship of complex II-supported respiration by isolated skeletal muscle mitochondria to ADP concentrations. We observed that O2 flux peaked at low ADP concentration ([ADP]) (high ΔΨ) before declining at higher [ADP] (low ΔΨ). The decline resulted from oxaloacetate (OAA) accumulation and inhibition of succinate dehydrogenase. This prompted us to question the effect of incremental [ADP] on respiration in interscapular brown adipose tissue (IBAT) mitochondria, wherein ΔΨ is intrinsically low because of uncoupling protein 1 (UCP1). We found that succinate-energized IBAT mitochondria, even in the absence of ADP, accumulate OAA and manifest limited respiration, similar to muscle mitochondria at high [ADP]. This could be prevented by guanosine 5'-diphosphate inhibition of UCP1. NAD+ cycling with NADH requires complex I electron flow and is needed to form OAA. Therefore, to assess the role of electron transit, we perturbed flow using a small molecule, N1-(3-acetamidophenyl)-N2-(2-(4-methyl-2-(p-tolyl)thiazol-5-yl)ethyl)oxalamide. We observed decreased OAA, increased NADH/NAD+, and increased succinate-supported mitochondrial respiration under conditions of low ΔΨ (IBAT) but not high ΔΨ (heart). In summary, complex II-energized respiration in IBAT mitochondria is tempered by complex I-derived OAA in a manner dependent on UCP1. These dynamics depend on electron transit in complex I.-Fink, B. D., Yu, L., Sivitz, W. I. Modulation of complex II-energized respiration in muscle, heart, and brown adipose mitochondria by oxaloacetate and complex I electron flow.

Keywords:  S1QEL 1.1; brown adipose tissue; mitochondrial respiratory chain; succinate dehydrogenase

DOI:  https://doi.org/10.1096/fj.201900690R
Cell Res. 2019 Jul 31.

NDUFAB1 confers cardio-protection by enhancing mitochondrial bioenergetics through coordination of respiratory complex and supercomplex assembly.

Tingting Hou, Rufeng Zhang, Chongshu Jian, Wanqiu Ding, Yanru Wang, Shukuan Ling, Qi Ma, Xinli Hu, Heping Cheng, Xianhua Wang.

The impairment of mitochondrial bioenergetics, often coupled with exaggerated reactive oxygen species (ROS) production, is a fundamental disease mechanism in organs with a high demand for energy, including the heart. Building a more robust and safer cellular powerhouse holds the promise for protecting these organs in stressful conditions. Here, we demonstrate that NADH:ubiquinone oxidoreductase subunit AB1 (NDUFAB1), also known as mitochondrial acyl carrier protein, acts as a powerful cardio-protector by conferring greater capacity and efficiency of mitochondrial energy metabolism. In particular, NDUFAB1 not only serves as a complex I subunit, but also coordinates the assembly of respiratory complexes I, II, and III, and supercomplexes, through regulating iron-sulfur biosynthesis and complex I subunit stability. Cardiac-specific deletion of Ndufab1 in mice caused defective bioenergetics and elevated ROS levels, leading to progressive dilated cardiomyopathy and eventual heart failure and sudden death. Overexpression of Ndufab1 effectively enhanced mitochondrial bioenergetics while limiting ROS production and protected the heart against ischemia-reperfusion injury. Together, our findings identify that NDUFAB1 is a crucial regulator of mitochondrial energy and ROS metabolism through coordinating the assembly of respiratory complexes and supercomplexes, and thus provide a potential therapeutic target for the prevention and treatment of heart failure.

DOI: https://doi.org/10.1038/s41422-019-0208-x
Cancers (Basel). 2019 Jul 30. pii: E1077. [Epub ahead of print]11(8):

Linking Cancer Metabolic Dysfunction and Genetic Instability through the Lens of Iron Metabolism.

Michael S Petronek, Douglas R Spitz, Garry R Buettner, Bryan G Allen.

  Iron (Fe) is an essential element that plays a fundamental role in a wide range of cellular functions, including cellular proliferation, DNA synthesis, as well as DNA damage and repair. Because of these connections, iron has been strongly implicated in cancer development. Cancer cells frequently have changes in the expression of iron regulatory proteins. For example, cancer cells frequently upregulate transferrin (increasing uptake of iron) and down regulate ferroportin (decreasing efflux of intracellular iron). These changes increase the steady-state level of intracellular redox active iron, known as the labile iron pool (LIP). The LIP typically contains approximately 2% intracellular iron, which primarily exists as ferrous iron (Fe2+). The LIP can readily contribute to oxidative distress within the cell through Fe2+-dioxygen and Fenton chemistries, generating the highly reactive hydroxyl radical (HO•). Due to the reactive nature of the LIP, it can contribute to increased DNA damage. Mitochondrial dysfunction in cancer cells results in increased steady-state levels of hydrogen peroxide and superoxide along with other downstream reactive oxygen species. The increased presence of H2O2 and O2•- can increase the LIP, contributing to increased mitochondrial uptake of iron as well as genetic instability. Thus, iron metabolism and labile iron pools may play a central role connecting the genetic mutational theories of cancer to the metabolic theories of cancer.

Keywords:  cancer; ferritin; genetic instability; genetic theory of cancer; iron metabolism; labile iron pool; metabolism; mitochondria; mitochondrial iron; transferrin receptors

DOI:  https://doi.org/10.3390/cancers11081077
Curr Opin Clin Nutr Metab Care. 2019 Sep;22(5): 347-354

Should we consider subcellular compartmentalization of metabolites, and if so, how do we measure them?

Kathryn E Wellen, Nathaniel W Snyder.

PURPOSE OF REVIEW: To examine the consequences of metabolism compartmentalized at the subcellular level, provide prototypical examples of compartmentalized metabolism, and describe methods to examine compartmentalized metabolism.
RECENT FINDINGS: Progress in metabolomics and isotope tracing has underscored the importance of subcellular compartments of metabolism. The discovery of biological effects of metabolites as bioenergetic intermediates, anabolic building blocks, signaling mediators, and effectors in posttranslation modifications of proteins and nucleic acids have highlighted the role of compartmentalization in determining metabolic fate. Recent advances in both direct and indirect methods to quantify compartmentalized metabolism have improved upon historical approaches. Genetically encoded metabolite sensors, chemical probes, immunoaffinity purification, and compartment-resolved metabolic modeling have all been recently applied to study compartmentalization.
SUMMARY: Accurate measurement of metabolites in distinct subcellular compartments is important for understanding and pharmacologically targeting metabolic pathways in diverse disease contexts, including cancer, diabetes, heart failure, obesity, and regulation of the immune system. Direct and indirect approaches to quantify compartmentalized metabolism are advancing rapidly. Yet, major challenges remain in the generalizability, rigor, and interpretation of data from the available methods to quantify compartmentalized metabolism.

DOI: https://doi.org/10.1097/MCO.0000000000000580
Sci Rep. 2019 Aug 02. 9(1): 11238

Quantitative Profiling of Oncometabolites in Frozen and Formalin-Fixed Paraffin-Embedded Tissue Specimens by Liquid Chromatography Coupled with Tandem Mass Spectrometry.

Xun Bao, Jianmei Wu, Brian Shuch, Patricia LoRusso, Ranjit S Bindra, Jing Li.

Given the implications of oncometabolites [succinate, fumarate, and 2-hydroxyglutarate (2HG)] in cancer pathogenesis and therapeutics, quantitative determination of their tissue levels has significant diagnostic, prognostic, and therapeutic values. Here, we developed and validated a multiplex liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) platform that allows simultaneous determination of oncometabolites (including succinate, fumarate and total 2HG) and other tricarboxylic acid cycle metabolites (α-ketoglutarate, malic acid, and glutamate) in frozen and FFPE tissues specimens. In addition, by employing chiral derivatization in the sample preparation, the platform enabled separation and determination of 2HG enantiomers (D- and L-2HG) in frozen and FFPE tissues. Isotope-labeled internal standard method was used for the quantitation. Linear calibration curve ranges in aqueous solution were 0.02-10, 0.2-100, 0.002-10, and 0.002-5 µM for succinate, fumarate, total 2HG, and D/L-2HG, respectively. Intra- and inter-day precision and accuracy for individual oncometabolites were within the generally accepted criteria for bioanalytical method validation (<15%). The recovery of spiked individual oncometabolites from pooled homogenate of FFPE or frozen tissue ranged 86-112%. Method validation indicated the technical feasibility, reliability and reproducibility of the platform. Oncometabolites were notably lost during the routine FFPE process. The ratios of succinate to glutamate, fumarate to α-ketoglutarate, 2HG to glutamate, and D-2HG to L-2HG were reliable surrogate measurements for the detection of altered levels of oncometabolites in FFPE specimens. Our study laid a foundation for the utility of archival FFPE specimens for oncometabolite profiling as a valid technique in clinical research and routine medical care.

DOI: https://doi.org/10.1038/s41598-019-47669-5
Nat Cell Biol. 2019 Aug;21(8): 978-990

5-methylcytosine promotes pathogenesis of bladder cancer through stabilizing mRNAs.

Xin Chen, Ang Li, Bao-Fa Sun, Ying Yang, Ya-Nan Han, Xun Yuan, Ri-Xin Chen, Wen-Su Wei, Yanchao Liu, Chun-Chun Gao, Yu-Sheng Chen, Mengmeng Zhang, Xiao-Dan Ma, Zhuo-Wei Liu, Jun-Hang Luo, Cong Lyu, Hai-Lin Wang, Jinbiao Ma, Yong-Liang Zhao, Fang-Jian Zhou, Ying Huang, Dan Xie, Yun-Gui Yang.

Although 5-methylcytosine (m5C) is a widespread modification in RNAs, its regulation and biological role in pathological conditions (such as cancer) remain unknown. Here, we provide the single-nucleotide resolution landscape of messenger RNA m5C modifications in human urothelial carcinoma of the bladder (UCB). We identify numerous oncogene RNAs with hypermethylated m5C sites causally linked to their upregulation in UCBs and further demonstrate YBX1 as an m5C 'reader' recognizing m5C-modified mRNAs through the indole ring of W65 in its cold-shock domain. YBX1 maintains the stability of its target mRNA by recruiting ELAVL1. Moreover, NSUN2 and YBX1 are demonstrated to drive UCB pathogenesis by targeting the m5C methylation site in the HDGF 3' untranslated region. Clinically, a high coexpression of NUSN2, YBX1 and HDGF predicts the poorest survival. Our findings reveal an unprecedented mechanism of RNA m5C-regulated oncogene activation, providing a potential therapeutic strategy for UCB.

DOI: https://doi.org/10.1038/s41556-019-0361-y
SLAS Discov. 2019 Aug 02. 2472555219864894

Targeting the Mitochondrial Potassium Channel Kv1.3 to Kill Cancer Cells: Drugs, Strategies, and New Perspectives.

Elena Prosdocimi, Vanessa Checchetto, Luigi Leanza.

  Cancer is the consequence of aberrations in cell growth or cell death. In this scenario, mitochondria and ion channels play a critical role in regard to cell proliferation, malignant angiogenesis, migration, and metastasis. In this review, we focus on Kv1.3 and specifically on mitoKv1.3, which showed an aberrant expression in cancer cells compared with healthy tissues and which is involved in the apoptotic pathway. In recent years, mitoKv1.3 has become an oncological target since its pharmacological modulation has been demonstrated to reduce tumor growth and progression both in vitro and in vivo using preclinical mouse models of different types of tumors.

Keywords:  cancer; ion channels; mitochondria

DOI:  https://doi.org/10.1177/2472555219864894
Nat Commun. 2019 Jul 29. 10(1): 3390

Mitofusins regulate lipid metabolism to mediate the development of lung fibrosis.

Kuei-Pin Chung, Chia-Lang Hsu, Li-Chao Fan, Ziling Huang, Divya Bhatia, Yi-Jung Chen, Shu Hisata, Soo Jung Cho, Kiichi Nakahira, Mitsuru Imamura, Mary E Choi, Chong-Jen Yu, Suzanne M Cloonan, Augustine M K Choi.

Accumulating evidence illustrates a fundamental role for mitochondria in lung alveolar type 2 epithelial cell (AEC2) dysfunction in the pathogenesis of idiopathic pulmonary fibrosis. However, the role of mitochondrial fusion in AEC2 function and lung fibrosis development remains unknown. Here we report that the absence of the mitochondrial fusion proteins mitofusin1 (MFN1) and mitofusin2 (MFN2) in murine AEC2 cells leads to morbidity and mortality associated with spontaneous lung fibrosis. We uncover a crucial role for MFN1 and MFN2 in the production of surfactant lipids with MFN1 and MFN2 regulating the synthesis of phospholipids and cholesterol in AEC2 cells. Loss of MFN1, MFN2 or inhibiting lipid synthesis via fatty acid synthase deficiency in AEC2 cells exacerbates bleomycin-induced lung fibrosis. We propose a tenet that mitochondrial fusion and lipid metabolism are tightly linked to regulate AEC2 cell injury and subsequent fibrotic remodeling in the lung.

DOI: https://doi.org/10.1038/s41467-019-11327-1
Eur J Immunol. 2019 Aug 02.

Mitochondrial functionality and metabolism in T cells from progressive multiple sclerosis patients.

Sara De Biasi, Anna Maria Simone, Elena Bianchini, Domenico Lo Tartaro, Simone Pecorini, Milena Nasi, Simone Patergnani, Gianluca Carnevale, Lara Gibellini, Diana Ferraro, Francesca Vitetta, Paolo Pinton, Patrizia Sola, Andrea Cossarizza, Marcello Pinti.

  Patients with primary progressive (PP) and secondary progressive (SP) forms of multiple sclerosis (MS) exhibit a sustained increase in the number of Th1, T cytotoxic type-1 and Th17 cells in peripheral blood, suggesting that the progressive phase is characterized by a permanent peripheral immune activation. As T cell functionality and activation are strictly connected to their metabolic profile, we investigated the mitochondrial functionality and metabolic changes of T cell subpopulations in a cohort of progressive MS patients. T cells from progressive patients were characterized by low proliferation and increase of terminally differentiated/exhausted cells. T cells from PP patients showed lower Oxygen Consumption Rate and Extracellular Acidification Rate, lower mitochondrial mass, membrane potential and respiration than those of SP patients, a downregulation of transcription factors supporting respiration and higher tendency to shift towards glycolysis upon stimulation. Furthermore, PP effector memory T cells were characterized by higher Glucose transporter -1 levels and a higher expression of glycolytic-supporting genes if compared to SP patients. Overall, our data suggest that profound differences exist in the phenotypic and metabolic features of T cells from PP and SP patients, even though the two clinical phenotypes are considered part of the same disease spectrum. This article is protected by copyright. All rights reserved.

Keywords:  Flow cytometry; Metabolism; Mitochondria; Multiple sclerosis; T cells

DOI:  https://doi.org/10.1002/eji.201948223
Nat Commun. 2019 Aug 02. 10(1): 3485

MYC paralog-dependent apoptotic priming orchestrates a spectrum of vulnerabilities in small cell lung cancer.

Marcel A Dammert, Johannes Brägelmann, Rachelle R Olsen, Stefanie Böhm, Niloufar Monhasery, Christopher P Whitney, Milind D Chalishazar, Hannah L Tumbrink, Matthew R Guthrie, Sebastian Klein, Abbie S Ireland, Jeremy Ryan, Anna Schmitt, Annika Marx, Luka Ozretić, Roberta Castiglione, Carina Lorenz, Ron D Jachimowicz, Elmar Wolf, Roman K Thomas, John T Poirier, Reinhard Büttner, Triparna Sen, Lauren A Byers, H Christian Reinhardt, Anthony Letai, Trudy G Oliver, Martin L Sos.

MYC paralogs are frequently activated in small cell lung cancer (SCLC) but represent poor drug targets. Thus, a detailed mapping of MYC-paralog-specific vulnerabilities may help to develop effective therapies for SCLC patients. Using a unique cellular CRISPR activation model, we uncover that, in contrast to MYCN and MYCL, MYC represses BCL2 transcription via interaction with MIZ1 and DNMT3a. The resulting lack of BCL2 expression promotes sensitivity to cell cycle control inhibition and dependency on MCL1. Furthermore, MYC activation leads to heightened apoptotic priming, intrinsic genotoxic stress and susceptibility to DNA damage checkpoint inhibitors. Finally, combined AURK and CHK1 inhibition substantially prolongs the survival of mice bearing MYC-driven SCLC beyond that of combination chemotherapy. These analyses uncover MYC-paralog-specific regulation of the apoptotic machinery with implications for genotype-based selection of targeted therapeutics in SCLC patients.

DOI: https://doi.org/10.1038/s41467-019-11371-x
Aging Cell. 2019 Aug 01. e13014

Hypothalamic mTORC2 is essential for metabolic health and longevity.

Karthikeyani Chellappa, Jacqueline A Brinkman, Sarmistha Mukherjee, Mark Morrison, Mohammed I Alotaibi, Kathryn A Carbajal, Amber L Alhadeff, Isaac J Perron, Rebecca Yao, Cole S Purdy, Denise M DeFelice, Matthew H Wakai, Jay Tomasiewicz, Amy Lin, Emma Meyer, Yajing Peng, Sebastian I Arriola Apelo, Luigi Puglielli, J Nicholas Betley, Georgios K Paschos, Joseph A Baur, Dudley W Lamming.

  The mechanistic target of rapamycin (mTOR) is an evolutionarily conserved protein kinase that regulates growth and metabolism. mTOR is found in two protein complexes, mTORC1 and mTORC2, that have distinct components and substrates and are both inhibited by rapamycin, a macrolide drug that robustly extends lifespan in multiple species including worms and mice. Although the beneficial effect of rapamycin on longevity is generally attributed to reduced mTORC1 signaling, disruption of mTORC2 signaling can also influence the longevity of worms, either positively or negatively depending on the temperature and food source. Here, we show that loss of hypothalamic mTORC2 signaling in mice decreases activity level, increases the set point for adiposity, and renders the animals susceptible to diet-induced obesity. Hypothalamic mTORC2 signaling normally increases with age, and mice lacking this pathway display higher fat mass and impaired glucose homeostasis throughout life, become more frail with age, and have decreased overall survival. We conclude that hypothalamic mTORC2 is essential for the normal metabolic health, fitness, and lifespan of mice. Our results have implications for the use of mTORC2-inhibiting pharmaceuticals in the treatment of brain cancer and diseases of aging.

Keywords:  frailty; hypothalamus; lifespanobesity; mTOR; mTORC2; obesity

DOI:  https://doi.org/10.1111/acel.13014
Cell Rep. 2019 Jul 30. pii: S2211-1247(19)30886-1. [Epub ahead of print]28(5): 1136-1143.e4

Glutamine Synthetase Promotes Radiation Resistance via Facilitating Nucleotide Metabolism and Subsequent DNA Damage Repair.

Shujun Fu, Zhi Li, Lanbo Xiao, Wenfeng Hu, Lu Zhang, Bowen Xie, Qin Zhou, Junju He, Yanfang Qiu, Ming Wen, Yanni Peng, Jie Gao, Rong Tan, Yuezhen Deng, Liang Weng, Lun-Quan Sun.

  Radiation resistance is a critical problem in radiotherapy for cancer. Radiation kills tumor cells mainly through causing DNA damage. Thus, efficiency of DNA damage repair is one of the most important factors that limits radiotherapy efficacy. Glutamine physiologically functions to generate protein and nucleotides. Here, we study the impact of glutamine metabolism on cancer therapeutic responses, in particular under irradiation-induced stress. We show that radiation-resistant cells possessed low glycolysis, mitochondrial respiration, and TCA cycle but high glutamine anabolism. Transcriptome analyses revealed that glutamine synthetase (GS), an enzyme catalyzing glutamate and ammonia to glutamine, was responsible for the metabolic alteration. ChIP and luciferase reporter assays revealed that GS could be transcriptionally regulated by STAT5. Knockdown of GS delayed DNA repair, weakened nucleotide metabolism, and enhanced radiosensitivity both in vitro and in vivo. Our data show that GS links glutamine metabolism to radiotherapy response through fueling nucleotide synthesis and accelerating DNA repair.

Keywords:  DNA damage repair; glutamine synthetase; radiation resistance

DOI:  https://doi.org/10.1016/j.celrep.2019.07.002
Cell Metab. 2019 Jul 18. pii: S1550-4131(19)30375-4. [Epub ahead of print]

Targeting Pathogenic Lafora Bodies in Lafora Disease Using an Antibody-Enzyme Fusion.

M Kathryn Brewer, Annette Uittenbogaard, Grant L Austin, Dyann M Segvich, Anna DePaoli-Roach, Peter J Roach, John J McCarthy, Zoe R Simmons, Jason A Brandon, Zhengqiu Zhou, Jill Zeller, Lyndsay E A Young, Ramon C Sun, James R Pauly, Nadine M Aziz, Bradley L Hodges, Tracy R McKnight, Dustin D Armstrong, Matthew S Gentry.

  Lafora disease (LD) is a fatal childhood epilepsy caused by recessive mutations in either the EPM2A or EPM2B gene. A hallmark of LD is the intracellular accumulation of insoluble polysaccharide deposits known as Lafora bodies (LBs) in the brain and other tissues. In LD mouse models, genetic reduction of glycogen synthesis eliminates LB formation and rescues the neurological phenotype. Therefore, LBs have become a therapeutic target for ameliorating LD. Herein, we demonstrate that human pancreatic α-amylase degrades LBs. We fused this amylase to a cell-penetrating antibody fragment, and this antibody-enzyme fusion (VAL-0417) degrades LBs in vitro and dramatically reduces LB loads in vivo in Epm2a-/- mice. Using metabolomics and multivariate analysis, we demonstrate that VAL-0417 treatment of Epm2a-/- mice reverses the metabolic phenotype to a wild-type profile. VAL-0417 is a promising drug for the treatment of LD and a putative precision therapy platform for intractable epilepsy.

Keywords:  Lafora bodies; Lafora disease; amylase; antibody-based drug; antibody-enzyme fusion; enzyme therapy; epilepsy; glycogen; glycogen storage disease; metabolomics

DOI:  https://doi.org/10.1016/j.cmet.2019.07.002
Cell. 2019 Jul 18. pii: S0092-8674(19)30686-5. [Epub ahead of print]

Repressive Gene Regulation Synchronizes Development with Cellular Metabolism.

Justin J Cassidy, Sebastian M Bernasek, Rachael Bakker, Ritika Giri, Nicolás Peláez, Bryan Eder, Anna Bobrowska, Neda Bagheri, Luis A Nunes Amaral, Richard W Carthew.

  Metabolic conditions affect the developmental tempo of animals. Developmental gene regulatory networks (GRNs) must therefore synchronize their dynamics with a variable timescale. We find that layered repression of genes couples GRN output with variable metabolism. When repressors of transcription or mRNA and protein stability are lost, fewer errors in Drosophila development occur when metabolism is lowered. We demonstrate the universality of this phenomenon by eliminating the entire microRNA family of repressors and find that development to maturity can be largely rescued when metabolism is reduced. Using a mathematical model that replicates GRN dynamics, we find that lowering metabolism suppresses the emergence of developmental errors by curtailing the influence of auxiliary repressors on GRN output. We experimentally show that gene expression dynamics are less affected by loss of repressors when metabolism is reduced. Thus, layered repression provides robustness through error suppression and may provide an evolutionary route to a shorter reproductive cycle.

Keywords:  Drosophila; control theory; development; mathematical modeling; metabolism; microRNA

DOI:  https://doi.org/10.1016/j.cell.2019.06.023
PLoS Genet. 2019 Jul;15(7): e1008240

Defects of mitochondrial RNA turnover lead to the accumulation of double-stranded RNA in vivo.

Aleksandra Pajak, Isabelle Laine, Paula Clemente, Najla El-Fissi, Florian A Schober, Camilla Maffezzini, Javier Calvo-Garrido, Rolf Wibom, Roberta Filograna, Ashish Dhir, Anna Wedell, Christoph Freyer, Anna Wredenberg.

The RNA helicase SUV3 and the polynucleotide phosphorylase PNPase are involved in the degradation of mitochondrial mRNAs but their roles in vivo are not fully understood. Additionally, upstream processes, such as transcript maturation, have been linked to some of these factors, suggesting either dual roles or tightly interconnected mechanisms of mitochondrial RNA metabolism. To get a better understanding of the turn-over of mitochondrial RNAs in vivo, we manipulated the mitochondrial mRNA degrading complex in Drosophila melanogaster models and studied the molecular consequences. Additionally, we investigated if and how these factors interact with the mitochondrial poly(A) polymerase, MTPAP, as well as with the mitochondrial mRNA stabilising factor, LRPPRC. Our results demonstrate a tight interdependency of mitochondrial mRNA stability, polyadenylation and the removal of antisense RNA. Furthermore, disruption of degradation, as well as polyadenylation, leads to the accumulation of double-stranded RNAs, and their escape out into the cytoplasm is associated with an altered immune-response in flies. Together our results suggest a highly organised and inter-dependable regulation of mitochondrial RNA metabolism with far reaching consequences on cellular physiology.

DOI: https://doi.org/10.1371/journal.pgen.1008240
iScience. 2019 Jul 16. pii: S2589-0042(19)30241-X. [Epub ahead of print]19 83-92

The Circadian Protein Nocturnin Regulates Metabolic Adaptation in Brown Adipose Tissue.

Yasemin Onder, Isara Laothamatas, Stefano Berto, Katharina Sewart, Gokhul Kilaru, Bogdan Bordieanu, Jeremy J Stubblefield, Genevieve Konopka, Prashant Mishra, Carla B Green.

  Fine-tuning of transcriptional responses can be critical for long-term outcomes in response to an environmental challenge. The circadian protein Nocturnin belongs to a family of proteins that include exonucleases, endonucleases, and phosphatases and is most closely related to the CCR4 family of deadenylases that regulate the cellular transcriptome via control of poly(A) tail length of RNA transcripts. In this study, we investigate the role of Nocturnin in regulating the transcriptional response and downstream metabolic adaptations during cold exposure in brown adipose tissue. We find that Nocturnin exhibits dual localization within the cytosol and mitochondria, and loss of Nocturnin causes changes in expression of networks of mRNAs involved in mitochondrial function. Furthermore, Nocturnin-/- animals display significantly elevated levels of tricarboxylic acid cycle intermediates, indicating that they have distinct metabolic adaptations during a prolonged cold exposure. We conclude that cold-induced stimulation of Nocturnin levels can regulate long-term metabolic adaptations to environmental challenges.

Keywords:  Biological Sciences; Cell Biology; Metabolomics; Transcriptomics

DOI:  https://doi.org/10.1016/j.isci.2019.07.016
Cell Res. 2019 Jul 31.

De-fusing mitochondria defuses the mtDNA time-bomb.

Patrick F Chinnery, Julien Prudent.

DOI: https://doi.org/10.1038/s41422-019-0206-z
Cell Metab. 2019 Jul 19. pii: S1550-4131(19)30372-9. [Epub ahead of print]

Identification and Application of Gene Expression Signatures Associated with Lifespan Extension.

Alexander Tyshkovskiy, Perinur Bozaykut, Anastasia A Borodinova, Maxim V Gerashchenko, Gene P Ables, Michael Garratt, Philipp Khaitovich, Clary B Clish, Richard A Miller, Vadim N Gladyshev.

  Several pharmacological, dietary, and genetic interventions that increase mammalian lifespan are known, but general principles of lifespan extension remain unclear. Here, we performed RNA sequencing (RNA-seq) analyses of mice subjected to 8 longevity interventions. We discovered a feminizing effect associated with growth hormone regulation and diminution of sex-related differences. Expanding this analysis to 17 interventions with public data, we observed that many interventions induced similar gene expression changes. We identified hepatic gene signatures associated with lifespan extension across interventions, including upregulation of oxidative phosphorylation and drug metabolism, and showed that perturbed pathways may be shared across tissues. We further applied the discovered longevity signatures to identify new lifespan-extending candidates, such as chronic hypoxia, KU-0063794, and ascorbyl-palmitate. Finally, we developed GENtervention, an app that visualizes associations between gene expression changes and longevity. Overall, this study describes general and specific transcriptomic programs of lifespan extension in mice and provides tools to discover new interventions.

Keywords:  GENtervention; aging; caloric restriction; feminizing effect; gene expression; growth hormone; lifespan extension; lifespan-extending interventions; longevity; longevity signatures; rapamycin

DOI:  https://doi.org/10.1016/j.cmet.2019.06.018