bims-mepmim 2022-11-20 papers

bims-mepmim

Biomed News

on Metabolites in pathological microenvironments and immunometabolism

Issue of 2022‒11‒20
thirty-two papers selected by
Erika Mariana Palmieri
NIH/NCI Laboratory of Cancer ImmunoMetabolism

Targeting glutamine metabolism in hepatic stellate cells alleviates liver fibrosis.
Tumor cell-derived asymmetric dimethylarginine regulates macrophage functions and polarization.
The NLRP3 inflammasome: regulation by metabolic signals.
OXPHOS promotes apoptotic resistance and cellular persistence in TH17 cells in the periphery and tumor microenvironment.
A Tale of Two Lipids: Lipid Unsaturation Commands Ferroptosis Sensitivity.
CD8+ T cell metabolic rewiring defined by scRNA-seq identifies a critical role of ASNS expression dynamics in T cell differentiation.
Tracing the electron flow in redox metabolism: The appropriate distribution of electrons is essential to maintain redox balance in cancer cells.
Isolation of mouse mammary carcinoma-derived macrophages and cancer cells for co-culture assays.
ATP-binding cassette A1 and cholesterol transfer protein Aster-A promote an asymmetric cholesterol distribution in the plasma membrane.
Multiomics analysis couples mRNA turnover and translational control of glutamine metabolism to the differentiation of the activated CD4+ T cell.
Cancer metabolite blunts immunity.
Myeloid-derived itaconate suppresses cytotoxic CD8+ T cells and promotes tumour growth.
FABP5 controls macrophage alternative activation and allergic asthma by selectively programming long-chain unsaturated fatty acid metabolism.
Diabetic hyperglycemia promotes primary tumor progression through glycation-induced tumor extracellular matrix stiffening.
Mitochondrial dysfunction compromises ciliary homeostasis in astrocytes.
AMPK directly phosphorylates TBK1 to integrate glucose sensing into innate immunity.
Agr2-associated ER stress promotes adherent-invasive E. coli dysbiosis and triggers CD103+ dendritic cell IL-23-dependent ileocolitis.
Ribosome stalling is a signal for metabolic regulation by the ribotoxic stress response.
Molecular diversity of astrocytes.
Endogenous metabolism in endothelial and immune cells generates most of the tissue vitamin B3 (nicotinamide).
Chronic type I interferon signaling promotes lipid-peroxidation-driven terminal CD8+ T cell exhaustion and curtails anti-PD-1 efficacy.
Lactate supply overtakes glucose when neural computational and cognitive loads scale up.
The brain-fat connection.
Identification of purine biosynthesis as an NADH-sensing pathway to mediate energy stress.
Both ANT and ATPase are essential for mitochondrial permeability transition but not depolarization.
Intestinal plasticity and metabolism as regulators of organismal energy homeostasis.
Neutrophil breaching of the blood vessel pericyte layer during diapedesis requires mast cell-derived IL-17A.
Iron therapy mitigates chronic kidney disease progression by regulating intracellular iron status of kidney macrophages.
The adhesion G-protein-coupled receptor Gpr116 is essential to maintain the skeletal muscle stem cell pool.
Ciliary neurotrophic factor-mediated neuroprotection involves enhanced glycolysis and anabolism in degenerating mouse retinas.
Activation and signaling mechanism revealed by GPR119-Gs complex structures.
Altered glycolysis triggers impaired mitochondrial metabolism and mTORC1 activation in diabetic β-cells.

Cell Death Dis. 2022 Nov 14. 13(11): 955

Targeting glutamine metabolism in hepatic stellate cells alleviates liver fibrosis.

Xiaochun Yin, Jin Peng, Lihong Gu, Yan Liu, Xihan Li, Jinhui Wu, Bing Xu, Yuzheng Zhuge, Feng Zhang.

Glutamine metabolism plays an essential role in cell growth, and glutamate dehydrogenase (GDH) is a key enzyme. GDH promotes the metabolism of glutamate and glutamine to generate ATP, which is profoundly increased in multiple human cancers. Through in vitro and in vivo experiments, we verified that the small-molecule GDH inhibitor EGCG slowed the progression of fibrosis by inhibiting GDH enzyme activity and glutamine metabolism. SIRT4 is a mitochondrial enzyme with NAD that promotes ADP ribosylation and downregulates GDH activity. The role of SIRT4 in liver fibrosis and the related mechanisms are unknown. In this study, we measured the expression of SIRT4 and found that it was downregulated in liver fibrosis. Modest overexpression of SIRT4 protected the liver from fibrosis by inhibiting the transformation of glutamate to 2-ketoglutaric acid (α-KG) in the tricarboxylic acid cycle (TCA), thereby reducing the proliferative activity of hepatic stellate cells (HSCs). Collectively, our study reveals that SIRT4 controls GDH enzyme activity and expression, targeting glutamine metabolism in HSCs and alleviating liver fibrosis.

DOI: https://doi.org/10.1038/s41419-022-05409-0
Cancer Cell Int. 2022 Nov 15. 22(1): 351

Tumor cell-derived asymmetric dimethylarginine regulates macrophage functions and polarization.

Yi-Ling Chen, AKaychia T Lowery, Samuel Lin, Ameae M Walker, Kuan-Hui E Chen.

  BACKGROUND: Asymmetric dimethylarginine (ADMA), which is significantly elevated in the plasma of cancer patients, is formed via intracellular recycling of methylated proteins and serves as a precursor for resynthesis of arginine. However, the cause of ADMA elevation in cancers and its impact on the regulation of tumor immunity is not known.METHODS: Three mouse breast cell lines (normal breast epithelial HC11, breast cancer EMT6 and triple negative breast cancer 4T1) and their equivalent 3D stem cell culture were used to analyze the secretion of ADMA using ELISA and their responses to ADMA. Bone marrow-derived macrophages and/or RAW264.7 cells were used to determine the impact of increased extracellular ADMA on macrophage-tumor interactions. Gene/protein expression was analyzed through RNAseq, qPCR and flow cytometry. Protein functional analyses were conducted via fluorescent imaging (arginine uptake, tumor phagocytosis) and enzymatic assay (arginase activity). Cell viability was measured via MTS assay and/or direct cell counting using Countess III FL system.
RESULTS: For macrophages, ADMA impaired proliferation and phagocytosis of tumor cells, and even caused death in cultures incubated without arginine. ADMA also led to an unusual macrophage phenotype, with increased expression of arginase, cd163 and cd206 but decreased expression of il10 and dectin-1. In contrast to the severely negative impacts on macrophages, ADMA had relatively minor effects on proliferation and survival of mouse normal epithelial HC11 cells, mouse breast cancer EMT6 and 4T1 cells, but there was increased expression of the mesenchymal markers, vimentin and snail2, and decreased expression of the epithelial marker, mucin-1 in EMT6 cells. When tumor cells were co-cultured ex vivo with tumor antigen in vivo-primed splenocytes, the tumor cells secreted more ADMA and there were alterations in the tumor cell arginine metabolic landscape, including increased expression of genes involved in arginine uptake, metabolism and methylation, and decreased expression of a gene that is responsible for arginine demethylation. Additionally, interferon-gamma, a cytokine involved in immune challenge, increased secretion of ADMA in tumor cells, a process attenuated by an autophagy inhibitor.
CONCLUSION: Our results suggest initial immune attack promotes autophagy in tumor cells, which then secrete ADMA to manipulate macrophage polarization favoring tumor tolerance.

Keywords:  Arginine metabolism; Asymmetric dimethylarginine; Autophagy; Macrophage polarization; cancer stem cells

DOI:  https://doi.org/10.1186/s12935-022-02769-7
Trends Immunol. 2022 Nov 09. pii: S1471-4906(22)00211-3. [Epub ahead of print]

The NLRP3 inflammasome: regulation by metabolic signals.

Antoni Olona, Stuart Leishman, Paras K Anand.

  Macrophages undergo profound metabolic reprogramming upon sensing infectious and sterile stimuli. This metabolic shift supports and regulates essential innate immune functions, including activation of the NLRP3 inflammasome. Within distinct metabolic networks, key enzymes play pivotal roles to control flux restraining detrimental inflammasome signaling. However, depending on the metabolic cues, specific enzymes and metabolites result in inflammasome activation outcomes which contrast other metabolic steps in the pathway. We posit that understanding which metabolic steps commit to discrete inflammasome fates will broaden our understanding of metabolic checkpoints to maintain homeostasis and offer better therapeutic options in human disease.

Keywords:  NLRP3; TCA cycle; glycolysis; inflammasome; lipids; metabolism

DOI:  https://doi.org/10.1016/j.it.2022.10.003
Sci Immunol. 2022 Nov 25. 7(77): eabm8182

OXPHOS promotes apoptotic resistance and cellular persistence in TH17 cells in the periphery and tumor microenvironment.

Hanna S Hong, Nneka E Mbah, Mengrou Shan, Kristen Loesel, Lin Lin, Peter Sajjakulnukit, Luis O Correa, Anthony Andren, Jason Lin, Atsushi Hayashi, Brian Magnuson, Judy Chen, Zhaoheng Li, Yuying Xie, Li Zhang, Daniel R Goldstein, Shannon A Carty, Yu Leo Lei, Anthony W Opipari, Rafael J Argüello, Ilona Kryczek, Nobuhiko Kamada, Weiping Zou, Luigi Franchi, Costas A Lyssiotis.

T cell proliferation and cytokine production are bioenergetically and biosynthetically costly. The inability to meet these metabolic demands results in altered differentiation, accompanied by impaired effector function, and attrition of the immune response. Interleukin-17-producing CD4 T cells (TH17s) are mediators of host defense, autoimmunity, and antitumor immunity in the setting of adoptive T cell therapy. TH17s are long-lived cells that require mitochondrial oxidative phosphorylation (OXPHOS) for effector function in vivo. Considering that TH17s polarized under standardized culture conditions are predominately glycolytic, little is known about how OXPHOS regulates TH17 processes, such as their ability to persist and thus contribute to protracted immune responses. Here, we modified standardized culture medium and identified a culture system that reliably induces OXPHOS dependence in TH17s. We found that TH17s cultured under OXPHOS conditions metabolically resembled their in vivo counterparts, whereas glycolytic cultures were dissimilar. OXPHOS TH17s exhibited increased mitochondrial fitness, glutamine anaplerosis, and an antiapoptotic phenotype marked by high BCL-XL and low BIM. Limited mitophagy, mediated by mitochondrial fusion regulator OPA-1, was critical to apoptotic resistance in OXPHOS TH17s. By contrast, glycolytic TH17s exhibited more mitophagy and an imbalance in BCL-XL to BIM, thereby priming them for apoptosis. In addition, through adoptive transfer experiments, we demonstrated that OXPHOS protected TH17s from apoptosis while enhancing their persistence in the periphery and tumor microenvironment in a murine model of melanoma. Together, our work demonstrates how metabolism regulates TH17 cell fate and highlights the potential for therapies that target OXPHOS in TH17-driven diseases.

DOI: https://doi.org/10.1126/sciimmunol.abm8182
Proteomics. 2022 Nov 18. e2100308

A Tale of Two Lipids: Lipid Unsaturation Commands Ferroptosis Sensitivity.

Jason Rodencal, Scott J Dixon.

  Membrane lipids play important roles in the regulation of cell fate, including the execution of ferroptosis. Ferroptosis is a non-apoptotic cell death mechanism defined by iron-dependent membrane lipid peroxidation. Phospholipids containing polyunsaturated fatty acids (PUFAs) are highly vulnerable to peroxidation and are essential for ferroptosis execution. By contrast, the incorporation of less oxidizable monounsaturated fatty acids (MUFAs) in membrane phospholipids protects cells from ferroptosis. The enzymes and pathways that govern PUFA and MUFA metabolism therefore play a critical role in determining cellular sensitivity to ferroptosis. Here, we review three lipid metabolic processes fatty acid biosynthesis, ether lipid biosynthesis, and phospholipid remodeling-that govern ferroptosis sensitivity by regulating the balance of PUFAs and MUFAs in membrane phospholipids. This article is protected by copyright. All rights reserved.

Keywords:  PUFA; ether lipid; ferroptosis; iron; membrane; necrosis

DOI:  https://doi.org/10.1002/pmic.202100308
Cell Rep. 2022 Nov 15. pii: S2211-1247(22)01510-8. [Epub ahead of print]41(7): 111639

CD8+ T cell metabolic rewiring defined by scRNA-seq identifies a critical role of ASNS expression dynamics in T cell differentiation.

Juan Fernández-García, Fabien Franco, Sweta Parik, Patricia Altea-Manzano, Antonino Alejandro Pane, Dorien Broekaert, Joke van Elsen, Ines Vermeire, Tessa Schalley, Mélanie Planque, Thomas van Brussel, Rogier Schepers, Elodie Modave, Tobias K Karakach, Peter Carmeliet, Diether Lambrechts, Ping-Chih Ho, Sarah-Maria Fendt.

  T cells dynamically rewire their metabolism during an immune response. We applied single-cell RNA sequencing to CD8+ T cells activated and differentiated in vitro in physiological medium to resolve these metabolic dynamics. We identify a differential time-dependent reliance of activating T cells on the synthesis versus uptake of various non-essential amino acids, which we corroborate with functional assays. We also identify metabolic genes that potentially dictate the outcome of T cell differentiation, by ranking them based on their expression dynamics. Among them, we find asparagine synthetase (Asns), whose expression peaks for effector T cells and decays toward memory formation. Disrupting these expression dynamics by ASNS overexpression promotes an effector phenotype, enhancing the anti-tumor response of adoptively transferred CD8+ T cells in a mouse melanoma model. We thus provide a resource of dynamic expression changes during CD8+ T cell activation and differentiation, and identify ASNS expression dynamics as a modulator of CD8+ T cell differentiation.

Keywords:  ASNS; CD8(+) T cells; CP: Immunology; T-cell activation; T-cell differentiation; asparagine; dynamics; immunology; metabolism; physiological media; scRNA-seq

DOI:  https://doi.org/10.1016/j.celrep.2022.111639
Semin Cancer Biol. 2022 Oct 28. pii: S1044-579X(22)00209-7. [Epub ahead of print]87 32-47

Tracing the electron flow in redox metabolism: The appropriate distribution of electrons is essential to maintain redox balance in cancer cells.

Minfeng Ying, Xun Hu.

  Cancer cells are characterized by sustained proliferation, which requires a huge demand of fuels to support energy production and biosynthesis. Energy is produced by the oxidation of the fuels during catabolism, and biosynthesis is achieved by the reduction of smaller units or precursors. Therefore, the oxidation-reduction (redox) reactions in cancer cells are more active compared to those in the normal counterparts. The higher activity of redox metabolism also induces a more severe oxidative stress, raising the question of how cancer cells maintain the redox balance. In this review, we overview the redox metabolism of cancer cells in an electron-tracing view. The electrons are derived from the nutrients in the tumor microenvironment and released during catabolism. Most of the electrons are transferred to NAD(P) system and then directed to four destinations: energy production, ROS generation, reductive biosynthesis and antioxidant system. The appropriate distribution of these electrons achieved by the function of redox regulation network is essential to maintain redox homeostasis in cancer cells. Interfering with the electron distribution and disrupting redox balance by targeting the redox regulation network may provide therapeutic implications for cancer treatment.

Keywords:  Antioxidant system; Cancer metabolism; NADPH; ROS; Redox balance

DOI:  https://doi.org/10.1016/j.semcancer.2022.10.005
STAR Protoc. 2022 Dec 16. 3(4): 101833

Isolation of mouse mammary carcinoma-derived macrophages and cancer cells for co-culture assays.

Lijuan Sun, Xiao Han, Mikala Egeblad.

  We recently established an in vitro co-culture system in which monophosphoryl lipid A + interferon-γ (MPLA+IFNγ)-treated tumor-associated macrophages (TAMs) killed cancer cells. Here, we describe a step-by-step protocol for isolating TAMs and cancer cells from mouse primary mammary carcinomas, the setup of the co-culture system, and the image acquisition approach. The technical difficulties in the co-culture assay involve isolating pure TAMs and cancer cells from the same tumor and staining them with different dyes to track the macrophages' tumoricidal activity. For complete details on the use and execution of this protocol, please refer to Sun et al. (2021).1.

Keywords:  Cancer; Cell Biology; Cell culture; Cell isolation; Immunology; Microscopy; Molecular/Chemical Probes

DOI:  https://doi.org/10.1016/j.xpro.2022.101833
J Biol Chem. 2022 Nov 14. pii: S0021-9258(22)01145-0. [Epub ahead of print] 102702

ATP-binding cassette A1 and cholesterol transfer protein Aster-A promote an asymmetric cholesterol distribution in the plasma membrane.

Fumihiko Ogasawara, Kazumitsu Ueda.

  Cholesterol is a major and essential component of the mammalian cell plasma membrane (PM), and the loss of cholesterol homeostasis leads to various pathologies. Cellular cholesterol uptake and synthesis are regulated by a cholesterol sensor in the endoplasmic reticulum (ER). However, it remains unclear how changes in the cholesterol level of the PM are recognized. Here we show that the sensing of cholesterol in the PM depends on ATP-binding cassette A1 (ABCA1) and the cholesterol transfer protein Aster-A, which cooperatively maintain the asymmetric transbilayer cholesterol distribution in the PM. We demonstrate that ABCA1 translocates (flops) cholesterol from the inner leaflet of the PM to the outer leaflet to maintain a low inner leaflet cholesterol level. We also found when inner cholesterol levels were increased, Aster-A was recruited to the PM-ER contact site to transfer cholesterol to the ER. These results suggest that ABCA1 could promote an asymmetric cholesterol distribution to suppress Aster-A recruitment to the PM-ER contact site to maintain intracellular cholesterol homeostasis.

Keywords:  ATP-binding cassette A1 (ABCA1); Aster-A (GramD1a); PM-ER contact site; asymmetric transbilayer cholesterol distribution; cholesterol flop; intracellular cholesterol homeostasis

DOI:  https://doi.org/10.1016/j.jbc.2022.102702
Sci Rep. 2022 Nov 16. 12(1): 19657

Multiomics analysis couples mRNA turnover and translational control of glutamine metabolism to the differentiation of the activated CD4+ T cell.

Louise S Matheson, Georg Petkau, Beatriz Sáenz-Narciso, Vanessa D'Angeli, Jessica McHugh, Rebecca Newman, Haydn Munford, James West, Krishnendu Chakraborty, Jennie Roberts, Sebastian Łukasiak, Manuel D Díaz-Muñoz, Sarah E Bell, Sarah Dimeloe, Martin Turner.

The ZFP36 family of RNA-binding proteins acts post-transcriptionally to repress translation and promote RNA decay. Studies of genes and pathways regulated by the ZFP36 family in CD4+ T cells have focussed largely on cytokines, but their impact on metabolic reprogramming and differentiation is unclear. Using CD4+ T cells lacking Zfp36 and Zfp36l1, we combined the quantification of mRNA transcription, stability, abundance and translation with crosslinking immunoprecipitation and metabolic profiling to determine how they regulate T cell metabolism and differentiation. Our results suggest that ZFP36 and ZFP36L1 act directly to limit the expression of genes driving anabolic processes by two distinct routes: by targeting transcription factors and by targeting transcripts encoding rate-limiting enzymes. These enzymes span numerous metabolic pathways including glycolysis, one-carbon metabolism and glutaminolysis. Direct binding and repression of transcripts encoding glutamine transporter SLC38A2 correlated with increased cellular glutamine content in ZFP36/ZFP36L1-deficient T cells. Increased conversion of glutamine to α-ketoglutarate in these cells was consistent with direct binding of ZFP36/ZFP36L1 to Gls (encoding glutaminase) and Glud1 (encoding glutamate dehydrogenase). We propose that ZFP36 and ZFP36L1 as well as glutamine and α-ketoglutarate are limiting factors for the acquisition of the cytotoxic CD4+ T cell fate. Our data implicate ZFP36 and ZFP36L1 in limiting glutamine anaplerosis and differentiation of activated CD4+ T cells, likely mediated by direct binding to transcripts of critical genes that drive these processes.

DOI: https://doi.org/10.1038/s41598-022-24132-6
Nat Metab. 2022 Nov 18.

Cancer metabolite blunts immunity.

Michael Attwaters.

DOI: https://doi.org/10.1038/s42255-022-00690-x
Nat Metab. 2022 Nov 14.

Myeloid-derived itaconate suppresses cytotoxic CD8+ T cells and promotes tumour growth.

Hongyun Zhao, Da Teng, Lifeng Yang, Xincheng Xu, Jiajia Chen, Tengjia Jiang, Austin Y Feng, Yaqing Zhang, Dennie T Frederick, Lei Gu, Li Cai, John M Asara, Marina Pasca di Magliano, Genevieve M Boland, Keith T Flaherty, Kenneth D Swanson, David Liu, Joshua D Rabinowitz, Bin Zheng.

The tumour microenvironment possesses mechanisms that suppress anti-tumour immunity. Itaconate is a metabolite produced from the Krebs cycle intermediate cis-aconitate by the activity of immune-responsive gene 1 (IRG1). While it is known to be immune modulatory, the role of itaconate in anti-tumour immunity is unclear. Here, we demonstrate that myeloid-derived suppressor cells (MDSCs) secrete itaconate that can be taken up by CD8+ T cells and suppress their proliferation, cytokine production and cytolytic activity. Metabolite profiling, stable-isotope tracing and metabolite supplementation studies indicated that itaconate suppressed the biosynthesis of aspartate and serine/glycine in CD8+ T cells to attenuate their proliferation and function. Host deletion of Irg1 in female mice bearing allografted tumours resulted in decreased tumour growth, inhibited the immune-suppressive activities of MDSCs, promoted anti-tumour immunity of CD8+ T cells and enhanced the anti-tumour activity of anti-PD-1 antibody treatment. Furthermore, we found a significant negative correlation between IRG1 expression and response to PD-1 immune checkpoint blockade in patients with melanoma. Our findings not only reveal a previously unknown role of itaconate as an immune checkpoint metabolite secreted from MDSCs to suppress CD8+ T cells, but also establish IRG1 as a myeloid-selective target in immunometabolism whose inhibition promotes anti-tumour immunity and enhances the efficacy of immune checkpoint protein blockade.

DOI: https://doi.org/10.1038/s42255-022-00676-9
Cell Rep. 2022 Nov 15. pii: S2211-1247(22)01542-X. [Epub ahead of print]41(7): 111668

FABP5 controls macrophage alternative activation and allergic asthma by selectively programming long-chain unsaturated fatty acid metabolism.

Yangxiao Hou, Dong Wei, Zhaoqi Zhang, Han Guo, Sihong Li, Jiayu Zhang, Peng Zhang, Lianfeng Zhang, Yong Zhao.

  Fatty acids (FAs) are widely involved in diverse biological functions. In mice with myeloid-specific deletion of fatty acid-binding protein 5 (FABP5), OVA-induced allergic airway inflammation (AAI) is significantly exacerbated by increasing alternatively activated macrophages (M2). Fabp5 deficiency increases IL-4-induced M2 in vitro. In macrophages, Fabp5 deletion causes significant accumulation of free long-chain unsaturated FAs, such as oleic acid, but does not cause detectable changes to other groups of FAs. Interestingly, excessive uptake of oleic acid aggravates AAI pathogenesis, with increased M2 polarization in bronchoalveolar lavage fluid. Informatics and mechanistic studies indicate that Fabp5 deficiency may reprogram metabolic pathways by enhancing FA β oxidation, tricarboxylic acid cycle, and oxidative phosphorylation, in addition to producing more ATP through activation of the PPARγ signaling pathway, reshaping macrophages in favor of M2 polarization. These results emphasize the importance of FABP5 and oleic acid in AAI, suggesting preventive and therapeutic strategies for allergic asthma.

Keywords:  CP: Immunology; FABP5; M2 macrophages; PPARγ; allergic airway inflammation; long-chain unsaturated fatty acids

DOI:  https://doi.org/10.1016/j.celrep.2022.111668
Sci Adv. 2022 Nov 16. 8(46): eabo1673

Diabetic hyperglycemia promotes primary tumor progression through glycation-induced tumor extracellular matrix stiffening.

Wenjun Wang, Lauren A Hapach, Lauren Griggs, Kyra Smart, Yusheng Wu, Paul V Taufalele, Matthew M Rowe, Katherine M Young, Madison E Bates, Andrew C Johnson, Nicholas J Ferrell, Ambra Pozzi, Cynthia A Reinhart-King.

Diabetes mellitus is a complex metabolic disorder that is associated with an increased risk of breast cancer. Despite this correlation, the interplay between tumor progression and diabetes, particularly with regard to stiffening of the extracellular matrix, is still mechanistically unclear. Here, we established a murine model where hyperglycemia was induced before breast tumor development. Using the murine model, in vitro systems, and patient samples, we show that hyperglycemia increases tumor growth, extracellular matrix stiffness, glycation, and epithelial-mesenchymal transition of tumor cells. Upon inhibition of glycation or mechanotransduction in diabetic mice, these same metrics are reduced to levels comparable with nondiabetic tumors. Together, our study describes a novel biomechanical mechanism by which diabetic hyperglycemia promotes breast tumor progression via glycating the extracellular matrix. In addition, our work provides evidence that glycation inhibition is a potential adjuvant therapy for diabetic cancer patients due to the key role of matrix stiffening in both diseases.

DOI: https://doi.org/10.1126/sciadv.abo1673
J Cell Biol. 2023 Jan 02. pii: e202203019. [Epub ahead of print]222(1):

Mitochondrial dysfunction compromises ciliary homeostasis in astrocytes.

Olesia Ignatenko, Satu Malinen, Sofiia Rybas, Helena Vihinen, Joni Nikkanen, Aleksander Kononov, Eija S Jokitalo, Gulayse Ince-Dunn, Anu Suomalainen.

Astrocytes, often considered as secondary responders to neurodegeneration, are emerging as primary drivers of brain disease. Here we show that mitochondrial DNA depletion in astrocytes affects their primary cilium, the signaling organelle of a cell. The progressive oxidative phosphorylation deficiency in astrocytes induces FOXJ1 and RFX transcription factors, known as master regulators of motile ciliogenesis. Consequently, a robust gene expression program involving motile cilia components and multiciliated cell differentiation factors are induced. While the affected astrocytes still retain a single cilium, these organelles elongate and become remarkably distorted. The data suggest that chronic activation of the mitochondrial integrated stress response (ISRmt) in astrocytes drives anabolic metabolism and promotes ciliary elongation. Collectively, our evidence indicates that an active signaling axis involving mitochondria and primary cilia exists and that ciliary signaling is part of ISRmt in astrocytes. We propose that metabolic ciliopathy is a novel pathomechanism for mitochondria-related neurodegenerative diseases.

DOI: https://doi.org/10.1083/jcb.202203019
Mol Cell. 2022 Nov 08. pii: S1097-2765(22)01054-1. [Epub ahead of print]

AMPK directly phosphorylates TBK1 to integrate glucose sensing into innate immunity.

Qian Zhang, Shengduo Liu, Chen-Song Zhang, Qirou Wu, Xinyuan Yu, Ruyuan Zhou, Fansen Meng, Ailian Wang, Fei Zhang, Shasha Chen, Xiaojian Wang, Lei Li, Jun Huang, Yao-Wei Huang, Jian Zou, Jun Qin, Tingbo Liang, Xin-Hua Feng, Sheng-Cai Lin, Pinglong Xu.

  Nutrient sensing and damage sensing are two fundamental processes in living organisms. While hyperglycemia is frequently linked to diabetes-related vulnerability to microbial infection, how body glucose levels affect innate immune responses to microbial invasion is not fully understood. Here, we surprisingly found that viral infection led to a rapid and dramatic decrease in blood glucose levels in rodents, leading to robust AMPK activation. AMPK, once activated, directly phosphorylates TBK1 at S511, which triggers IRF3 recruitment and the assembly of MAVS or STING signalosomes. Consistently, ablation or inhibition of AMPK, knockin of TBK1-S511A, or increased glucose levels compromised nucleic acid sensing, while boosting AMPK-TBK1 cascade by AICAR or TBK1-S511E knockin improves antiviral immunity substantially in various animal models. Thus, we identify TBK1 as an AMPK substrate, reveal the molecular mechanism coupling a dual sensing of glucose and nuclei acids, and report its physiological necessity in antiviral defense.

Keywords:  AMPK; TBK1; antiviral immunity; cGAS-STING; coronavirus; glucose metabolism; hyperglycemia; innate immunity; viral infection; virus-host interactions

DOI:  https://doi.org/10.1016/j.molcel.2022.10.026
Cell Rep. 2022 Nov 15. pii: S2211-1247(22)01508-X. [Epub ahead of print]41(7): 111637

Agr2-associated ER stress promotes adherent-invasive E. coli dysbiosis and triggers CD103+ dendritic cell IL-23-dependent ileocolitis.

Monica Viladomiu, Manirath Khounlotham, Belgin Dogan, Svetlana F Lima, Ahmed Elsaadi, Emre Cardakli, Jim G Castellanos, Charles Ng, Jeremy Herzog, Alexi A Schoenborn, Melissa Ellermann, Bo Liu, Shiying Zhang, Ajay S Gulati, R Balfour Sartor, Kenneth W Simpson, Steven M Lipkin, Randy S Longman.

  Endoplasmic reticulum (ER) stress is associated with Crohn's disease (CD), but its impact on host-microbe interaction in disease pathogenesis is not well defined. Functional deficiency in the protein disulfide isomerase anterior gradient 2 (AGR2) has been linked with CD and leads to epithelial cell ER stress and ileocolitis in mice and humans. Here, we show that ileal expression of AGR2 correlates with mucosal Enterobactericeae abundance in human inflammatory bowel disease (IBD) and that Agr2 deletion leads to ER-stress-dependent expansion of mucosal-associated adherent-invasive Escherichia coli (AIEC), which drives Th17 cell ileocolitis in mice. Mechanistically, our data reveal that AIEC-induced epithelial cell ER stress triggers CD103+ dendritic cell production of interleukin-23 (IL-23) and that IL-23R is required for ileocolitis in Agr2-/- mice. Overall, these data reveal a specific and reciprocal interaction of the expansion of the CD pathobiont AIEC with ER-stress-associated ileocolitis and highlight a distinct cellular mechanism for IL-23-dependent ileocolitis.

Keywords:  AIEC; Agr2; CP: Immunology; CP: Microbiology; ER stress; IL-23; Th17; ileitis

DOI:  https://doi.org/10.1016/j.celrep.2022.111637
Cell Metab. 2022 Nov 08. pii: S1550-4131(22)00489-2. [Epub ahead of print]

Ribosome stalling is a signal for metabolic regulation by the ribotoxic stress response.

Goda Snieckute, Aitana Victoria Genzor, Anna Constance Vind, Laura Ryder, Mark Stoneley, Sébastien Chamois, René Dreos, Cathrine Nordgaard, Frederike Sass, Melanie Blasius, Aida Rodríguez López, Sólveig Hlín Brynjólfsdóttir, Kasper Langebjerg Andersen, Anne E Willis, Lisa B Frankel, Steen Seier Poulsen, David Gatfield, Zachary Gerhart-Hines, Christoffer Clemmensen, Simon Bekker-Jensen.

  Impairment of translation can lead to collisions of ribosomes, which constitute an activation platform for several ribosomal stress-surveillance pathways. Among these is the ribotoxic stress response (RSR), where ribosomal sensing by the MAP3K ZAKα leads to activation of p38 and JNK kinases. Despite these insights, the physiological ramifications of ribosomal impairment and downstream RSR signaling remain elusive. Here, we show that stalling of ribosomes is sufficient to activate ZAKα. In response to amino acid deprivation and full nutrient starvation, RSR impacts on the ensuing metabolic responses in cells, nematodes, and mice. The RSR-regulated responses in these model systems include regulation of AMPK and mTOR signaling, survival under starvation conditions, stress hormone production, and regulation of blood sugar control. In addition, ZAK-/- male mice present a lean phenotype. Our work highlights impaired ribosomes as metabolic signals and demonstrates a role for RSR signaling in metabolic regulation.

Keywords:  AMPK; FGF21; ZAK-alpha; amino acid starvation; mTOR; metabolic regulation; mouse models; ribosome collision; ribotoxic stress response

DOI:  https://doi.org/10.1016/j.cmet.2022.10.011
Science. 2022 11 04. 378(6619): 475-476

Molecular diversity of astrocytes.

Katherine T Baldwin.

The tissue environment influences astrocyte form and function in health and disease.

DOI: https://doi.org/10.1126/science.ade9249
iScience. 2022 Nov 18. 25(11): 105431

Endogenous metabolism in endothelial and immune cells generates most of the tissue vitamin B3 (nicotinamide).

Julianna D Zeidler, Claudia C S Chini, Karina S Kanamori, Sonu Kashyap, Jair M Espindola-Netto, Katie Thompson, Gina Warner, Fernanda S Cabral, Thais R Peclat, Lilian Sales Gomez, Sierra A Lopez, Miles K Wandersee, Renee A Schoon, Kimberly Reid, Keir Menzies, Felipe Beckedorff, Joel M Reid, Sebastian Brachs, Ralph G Meyer, Mirella L Meyer-Ficca, Eduardo Nunes Chini.

  In mammals, nicotinamide (NAM) is the primary NAD precursor available in circulation, a signaling molecule, and a precursor for methyl-nicotinamide (M-NAM) synthesis. However, our knowledge about how the body regulates tissue NAM levels is still limited. Here we demonstrate that dietary vitamin B3 partially regulates plasma NAM and NAM-derived metabolites, but not their tissue levels. We found that NAD de novo synthesis from tryptophan contributes to plasma and tissue NAM, likely by providing substrates for NAD-degrading enzymes. We also demonstrate that tissue NAM is mainly generated by endogenous metabolism and that the NADase CD38 is the main enzyme that produces tissue NAM. Tissue-specific CD38-floxed mice revealed that CD38 activity on endothelial and immune cells is the major contributor to tissue steady-state levels of NAM in tissues like spleen and heart. Our findings uncover the presence of different pools of NAM in the body and a central role for CD38 in regulating tissue NAM levels.

Keywords:  Biochemistry; Biological sciences; Immunology; Molecular biology

DOI:  https://doi.org/10.1016/j.isci.2022.105431
Cell Rep. 2022 Nov 15. pii: S2211-1247(22)01518-2. [Epub ahead of print]41(7): 111647

Chronic type I interferon signaling promotes lipid-peroxidation-driven terminal CD8+ T cell exhaustion and curtails anti-PD-1 efficacy.

Weixin Chen, Jia Ming Nickolas Teo, Siu Wah Yau, Melody Yee-Man Wong, Chun-Nam Lok, Chi-Ming Che, Asif Javed, Yuanhua Huang, Stephanie Ma, Guang Sheng Ling.

  Identifying signals that govern the differentiation of tumor-infiltrating CD8+ T cells (CD8+ TILs) toward exhaustion can improve current therapeutic approaches for cancer. Here, we show that type I interferons (IFN-Is) act as environmental cues, enhancing terminal CD8+ T cell exhaustion in tumors. We find enrichment of IFN-I-stimulated genes (ISGs) within exhausted CD8+ T cells (Tex cells) in patients across various cancer types, with heightened ISG levels correlating with poor response to immune checkpoint blockade (ICB) therapy. In preclinical models, CD8+ TILs devoid of IFN-I signaling develop less exhaustion features, provide better tumor control, and show greater response to ICB-mediated rejuvenation. Mechanistically, chronic IFN-I stimulation perturbs lipid metabolism and redox balance in Tex cells, leading to aberrant lipid accumulation and elevated oxidative stress. Collectively, these defects promote lipid peroxidation, which potentiates metabolic and functional exhaustion of Tex cells. Thus, cell-intrinsic IFN-I signaling regulates the extent of CD8+ TIL exhaustion and has important implications for immunotherapy.

Keywords:  CD8(+) T cell exhaustion; CP: Immunology; anti-PD-1 therapy resistance; chronic IFN-I signaling; dysregulated lipid metabolism

DOI:  https://doi.org/10.1016/j.celrep.2022.111647
Proc Natl Acad Sci U S A. 2022 Nov 22. 119(47): e2212004119

Lactate supply overtakes glucose when neural computational and cognitive loads scale up.

Yulia Dembitskaya, Charlotte Piette, Sylvie Perez, Hugues Berry, Pierre J Magistretti, Laurent Venance.

  Neural computational power is determined by neuroenergetics, but how and which energy substrates are allocated to various forms of memory engram is unclear. To solve this question, we asked whether neuronal fueling by glucose or lactate scales differently upon increasing neural computation and cognitive loads. Here, using electrophysiology, two-photon imaging, cognitive tasks, and mathematical modeling, we show that both glucose and lactate are involved in engram formation, with lactate supporting long-term synaptic plasticity evoked by high-stimulation load activity patterns and high attentional load in cognitive tasks and glucose being sufficient for less demanding neural computation and learning tasks. Indeed, we show that lactate is mandatory for demanding neural computation, such as theta-burst stimulation, while glucose is sufficient for lighter forms of activity-dependent long-term potentiation (LTP), such as spike timing-dependent plasticity (STDP). We find that subtle variations of spike number or frequency in STDP are sufficient to shift the on-demand fueling from glucose to lactate. Finally, we demonstrate that lactate is necessary for a cognitive task requiring high attentional load, such as the object-in-place task, and for the corresponding in vivo hippocampal LTP expression but is not needed for a less demanding task, such as a simple novel object recognition. Overall, these results demonstrate that glucose and lactate metabolism are differentially engaged in neuronal fueling depending on the complexity of the activity-dependent plasticity and behavior.

Keywords:  glucose; lactate; learning and memory; neuroenergetic; synaptic plasticity

DOI:  https://doi.org/10.1073/pnas.2212004119
Science. 2022 11 04. 378(6619): 485

The brain-fat connection.

Filipa Cardoso.

Type 2 innate lymphoid cells shape metabolism through a brain-body circuit.

DOI: https://doi.org/10.1126/science.ade2132
Nat Commun. 2022 Nov 17. 13(1): 7031

Identification of purine biosynthesis as an NADH-sensing pathway to mediate energy stress.

Ronghui Yang, Chuanzhen Yang, Lingdi Ma, Yiliang Zhao, Zihao Guo, Jing Niu, Qiaoyun Chu, Yingmin Ma, Binghui Li.

An enhanced NADH/NAD+ ratio, termed reductive stress, is associated with many diseases. However, whether a downstream sensing pathway exists to mediate pathogenic outcomes remains unclear. Here, we generate a soluble pyridine nucleotide transhydrogenase from Escherichia coli (EcSTH), which can elevate the NADH/NAD+ ratio and meantime reduce the NADPH/NADP+ ratio. Additionally, we fuse EcSTH with previously described LbNOX (a water-forming NADH oxidase from Lactobacillus brevis) to resume the NADH/NAD+ ratio. With these tools and by using genome-wide CRISPR/Cas9 library screens and metabolic profiling in mammalian cells, we find that accumulated NADH deregulates PRPS2 (Ribose-phosphate pyrophosphokinase 2)-mediated downstream purine biosynthesis to provoke massive energy consumption, and therefore, the induction of energy stress. Blocking purine biosynthesis prevents NADH accumulation-associated cell death in vitro and tissue injury in vivo. These results underscore the pathophysiological role of deregulated purine biosynthesis in NADH accumulation-associated disorders and demonstrate the utility of EcSTH in manipulating NADH/NAD+ and NADPH/NADP+.

DOI: https://doi.org/10.1038/s41467-022-34850-0
iScience. 2022 Nov 18. 25(11): 105447

Both ANT and ATPase are essential for mitochondrial permeability transition but not depolarization.

M A Neginskaya, S E Morris, E V Pavlov.

  An increase in permeability of the mitochondrial inner membrane, mitochondrial permeability transition (PT), is the central event responsible for cell death and tissue damage in conditions such as stroke and heart attack. PT is caused by the cyclosporin A (CSA)-dependent calcium-induced pore, the permeability transition pore (PTP). The molecular details of PTP are incompletely understood. We utilized holographic and fluorescent microscopy to assess the contribution of ATP synthase and adenine nucleotide translocator (ANT) toward PTP. In cells lacking either ATP synthase or ANT, we observed CSA-sensitive membrane depolarization, but not high-conductance PTP. In wild-type cells, calcium-induced CSA-sensitive depolarization preceded opening of PTP, which occurred only after nearly complete mitochondrial membrane depolarization. We propose that both ATP synthase and ANT are required for high-conductance PTP but not depolarization, which presumably occurs through activation of the low-conductance PT, which has a molecular nature that is different from both complexes.

Keywords:  Cell biology; Functional aspects of cell biology; Molecular biology

DOI:  https://doi.org/10.1016/j.isci.2022.105447
Nat Metab. 2022 Nov 17.

Intestinal plasticity and metabolism as regulators of organismal energy homeostasis.

Ozren Stojanović, Irene Miguel-Aliaga, Mirko Trajkovski.

The small intestine displays marked anatomical and functional plasticity that includes adaptive alterations in adult gut morphology, enteroendocrine cell profile and their hormone secretion, as well as nutrient utilization and storage. In this Perspective, we examine how shifts in dietary and environmental conditions bring about changes in gut size, and describe how the intestine adapts to changes in internal state, bowel resection and gastric bypass surgery. We highlight the critical importance of these intestinal remodelling processes in maintaining energy balance of the organism, and in protecting the metabolism of other organs. The intestinal resizing is supported by changes in the microbiota composition, and by activation of carbohydrate and fatty acid metabolism, which govern the intestinal stem cell proliferation, intestinal cell fate, as well as survivability of differentiated epithelial cells. The discovery that intestinal remodelling is part of the normal physiological adaptation to various triggers, and the potential for harnessing the reversible gut plasticity, in our view, holds extraordinary promise for developing therapeutic approaches against metabolic and inflammatory diseases.

DOI: https://doi.org/10.1038/s42255-022-00679-6
Nat Commun. 2022 Nov 17. 13(1): 7029

Neutrophil breaching of the blood vessel pericyte layer during diapedesis requires mast cell-derived IL-17A.

Régis Joulia, Idaira María Guerrero-Fonseca, Tamara Girbl, Jonathon A Coates, Monja Stein, Laura Vázquez-Martínez, Eleanor Lynam, James Whiteford, Michael Schnoor, David Voehringer, Axel Roers, Sussan Nourshargh, Mathieu-Benoit Voisin.

Neutrophil diapedesis is an immediate step following infections and injury and is driven by complex interactions between leukocytes and various components of the blood vessel wall. Here, we show that perivascular mast cells (MC) are key regulators of neutrophil behaviour within the sub-endothelial space of inflamed venules. Using confocal intravital microscopy, we observe directed abluminal neutrophil motility along pericyte processes towards perivascular MCs, a response that created neutrophil extravasation hotspots. Conversely, MC-deficiency and pharmacological or genetic blockade of IL-17A leads to impaired neutrophil sub-endothelial migration and breaching of the pericyte layer. Mechanistically, identifying MCs as a significant cellular source of IL-17A, we establish that MC-derived IL-17A regulates the enrichment of key effector molecules ICAM-1 and CXCL1 in nearby pericytes. Collectively, we identify a novel MC-IL-17A-pericyte axis as modulator of the final steps of neutrophil diapedesis, with potential translational implications for inflammatory disorders driven by increased neutrophil diapedesis.

DOI: https://doi.org/10.1038/s41467-022-34695-7
JCI Insight. 2022 Nov 17. pii: e159235. [Epub ahead of print]

Iron therapy mitigates chronic kidney disease progression by regulating intracellular iron status of kidney macrophages.

Edwin Patino, Divya Bhatia, Steven Z Vance, Ada Antypiuk, Rie Uni, Chantalle Campbell, Carlo G Castillo, Shahd Jaouni, Francesca Vinchi, Mary E Choi, Oleh Akchurin.

  Systemic iron metabolism is disrupted in chronic kidney disease (CKD). However, little is known about local kidney iron homeostasis and its role in kidney fibrosis. Kidney-specific effects of iron therapy in CKD also remain elusive. Here, we elucidate the role of macrophage iron status in kidney fibrosis and demonstrate that it is a potential therapeutic target. In CKD, kidney macrophages exhibited depletion of labile iron pool (LIP) and induction of transferrin receptor 1, indicating intracellular iron deficiency. Low LIP in kidney macrophages was associated with their defective antioxidant response and pro-inflammatory polarization. Repletion of LIP in kidney macrophages through knockout of ferritin heavy chain (Fth1) reduced oxidative stress and mitigated fibrosis. Similar to Fth1 knockout, iron dextran therapy, through replenishing macrophage LIP, reduced oxidative stress, decreased the production of pro-inflammatory cytokines, and alleviated kidney fibrosis. Interestingly, iron significantly decreased TGF-β expression and suppressed TGF-β-driven fibrotic response of macrophages. Iron dextran therapy and FtH suppression had an additive protective effect against fibrosis. Adoptive transfer of iron-loaded macrophages alleviated kidney fibrosis, confirming the protective effect of iron-replete macrophages in CKD. Thus, targeting intracellular iron deficiency of kidney macrophages in CKD can serve as a therapeutic opportunity to mitigate disease progression.

Keywords:  Chronic kidney disease; Fibrosis; Macrophages; Nephrology

DOI:  https://doi.org/10.1172/jci.insight.159235
Cell Rep. 2022 Nov 15. pii: S2211-1247(22)01516-9. [Epub ahead of print]41(7): 111645

The adhesion G-protein-coupled receptor Gpr116 is essential to maintain the skeletal muscle stem cell pool.

Charlotte Sénéchal, Ryo Fujita, Solène Jamet, Arhamatoulaye Maiga, Junio Dort, Zakaria Orfi, Nicolas A Dumont, Michel Bouvier, Colin Crist.

  Skeletal muscle is populated with a reservoir of quiescent muscle stem cells (MuSCs), which regenerate the tissue after injury. Here, we show that the adhesion G-protein-coupled receptor Gpr116 is essential for long-term maintenance of the MuSC pool. Quiescent MuSCs express high levels of Gpr116, which is rapidly downregulated upon MuSC activation. MuSCs deficient for Gpr116 exhibit progressive depletion over time and are defective in self-renewal. Adhesion G-protein-coupled receptors contain an agonistic peptide sequence, called the "Stachel" sequence, within their long N-terminal ectodomains. Stimulation of MuSCs with the GPR116 Stachel peptide delays MuSC activation and differentiation. Stachel peptide stimulation of GPR116 leads to strong interaction with β-arrestins. Stimulation of GPR116 increases the nuclear localization of β-arrestin1, where it interacts with cAMP response element binding protein to regulate gene expression. Altogether, we propose a model by which GPR116 maintains the MuSC pool via nuclear functions of β-arrestin1.

Keywords:  CP: Stem cell research; GPR116; adhesion G-protein-coupled receptor; muscle stem cell; regeneration; skeletal muscle; β-arrestin1

DOI:  https://doi.org/10.1016/j.celrep.2022.111645
Nat Commun. 2022 Nov 17. 13(1): 7037

Ciliary neurotrophic factor-mediated neuroprotection involves enhanced glycolysis and anabolism in degenerating mouse retinas.

Kun Do Rhee, Yanjie Wang, Johanna Ten Hoeve, Linsey Stiles, Thao Thi Thu Nguyen, Xiangmei Zhang, Laurent Vergnes, Karen Reue, Orian Shirihai, Dean Bok, Xian-Jie Yang.

Ciliary neurotrophic factor (CNTF) acts as a potent neuroprotective cytokine in multiple models of retinal degeneration. To understand mechanisms underlying its broad neuroprotective effects, we have investigated the influence of CNTF on metabolism in a mouse model of photoreceptor degeneration. CNTF treatment improves the morphology of photoreceptor mitochondria, but also leads to reduced oxygen consumption and suppressed respiratory chain activities. Molecular analyses show elevated glycolytic pathway gene transcripts and active enzymes. Metabolomics analyses detect significantly higher levels of ATP and the energy currency phosphocreatine, elevated glycolytic pathway metabolites, increased TCA cycle metabolites, lipid biosynthetic pathway intermediates, nucleotides, and amino acids. Moreover, CNTF treatment restores the key antioxidant glutathione to the wild type level. Therefore, CNTF significantly impacts the metabolic status of degenerating retinas by promoting aerobic glycolysis and augmenting anabolic activities. These findings reveal cellular mechanisms underlying enhanced neuronal viability and suggest potential therapies for treating retinal degeneration.

DOI: https://doi.org/10.1038/s41467-022-34443-x
Nat Commun. 2022 Nov 17. 13(1): 7033

Activation and signaling mechanism revealed by GPR119-Gs complex structures.

Yuxia Qian, Jiening Wang, Linlin Yang, Yanru Liu, Lina Wang, Wei Liu, Yun Lin, Hong Yang, Lixin Ma, Sheng Ye, Shan Wu, Anna Qiao.

Agonists selectively targeting cannabinoid receptor-like G-protein-coupled receptor (GPCR) GPR119 hold promise for treating metabolic disorders while avoiding unwanted side effects. Here we present the cryo-electron microscopy (cryo-EM) structures of the human GPR119-Gs signaling complexes bound to AR231453 and MBX-2982, two representative agonists reported for GPR119. The structures reveal a one-amino acid shift of the conserved proline residue of TM5 that forms an outward bulge, opening up a hydrophobic cavity between TM4 and TM5 at the middle of the membrane for its endogenous ligands-monounsaturated lipid metabolites. In addition, we observed a salt bridge between ICL1 of GPR119 and Gβs. Disruption of the salt bridge eliminates the cAMP production of GPR119, indicating an important role of Gβs in GPR119-mediated signaling. Our structures, together with mutagenesis studies, illustrate the conserved binding mode of the chemically different agonists, and provide insights into the conformational changes in receptor activation and G protein coupling.

DOI: https://doi.org/10.1038/s41467-022-34696-6
Nat Commun. 2022 Nov 14. 13(1): 6754

Altered glycolysis triggers impaired mitochondrial metabolism and mTORC1 activation in diabetic β-cells.

Elizabeth Haythorne, Matthew Lloyd, John Walsby-Tickle, Andrei I Tarasov, Jonas Sandbrink, Idoia Portillo, Raul Terron Exposito, Gregor Sachse, Malgorzata Cyranka, Maria Rohm, Patrik Rorsman, James McCullagh, Frances M Ashcroft.

Chronic hyperglycaemia causes a dramatic decrease in mitochondrial metabolism and insulin content in pancreatic β-cells. This underlies the progressive decline in β-cell function in diabetes. However, the molecular mechanisms by which hyperglycaemia produces these effects remain unresolved. Using isolated islets and INS-1 cells, we show here that one or more glycolytic metabolites downstream of phosphofructokinase and upstream of GAPDH mediates the effects of chronic hyperglycemia. This metabolite stimulates marked upregulation of mTORC1 and concomitant downregulation of AMPK. Increased mTORC1 activity causes inhibition of pyruvate dehydrogenase which reduces pyruvate entry into the tricarboxylic acid cycle and partially accounts for the hyperglycaemia-induced reduction in oxidative phosphorylation and insulin secretion. In addition, hyperglycaemia (or diabetes) dramatically inhibits GAPDH activity, thereby impairing glucose metabolism. Our data also reveal that restricting glucose metabolism during hyperglycaemia prevents these changes and thus may be of therapeutic benefit. In summary, we have identified a pathway by which chronic hyperglycaemia reduces β-cell function.

DOI: https://doi.org/10.1038/s41467-022-34095-x