Metabolic regulation of immune response in health and disease ”VACCINES AND VACCINATION” MOSCOW, SEPTEMBER 27-OCTOBER 1, 2017 Metabolic regulation of immune response in health and disease BIRKE BARTOSCH Cancer Center, Lyon, France

T cell activation is metabolically regulated Rapid proliferation effector Contraction memory Cell number pathogen resting …………………….. Glycolysis Glutaminolysis Nutrient uptake ……….. ……….. Time Pearce et al. Science 2013

Metabolism of T cell activation shares common features with Cancer metabolism Quiescent cell Glucose Fatty acids Glycolysis Pyruvate Lipids ATP (O2) TCA cycle Glutaminolysis Glutamine

Metabolism of T cell activation shares common features with Cancer metabolism Quiescent cell Proliferating cell Glucose Glucose Lipids Nucleotides NADPH, ATP Fatty acids Glycolysis Pyruvate Pyruvate Lactate Lipids ATP (O2) Lipids ATP (O2) TCA cycle TCA cycle Glutaminolysis Glutamine

Metabolism of T cell activation shares common features with Cancer metabolism Quiescent cell Proliferating cell Glucose Glucose Lipids Nucleotides NADPH, ATP Fatty acids Glycolysis Pyruvate Pyruvate Lactate Lipids ATP (O2) Lipids ATP (O2) TCA cycle TCA cycle AA, GSH Glutaminolysis Glutamine Glutamine

Glycolysis in T cell biology Roles of glycolysis in immune functions: Bioenergetic and anabolic roles Transcriptional regulation Post-transcriptional regulation Glycolytic intermediates as signaling molecules Targeting glycolysis to impact immune functions Glycolysis as target of immune invasion Systemic metabolic alterations Glutaminolysis in T cell biology

Bioenergetic and anabolic roles Glycolysis Bioenergetic and anabolic roles Glucose Glucose 6P Pentose phosphate shunt: Nucleic acids NADPH Fructose 6P Fructose 1,6biP Glyceraldehyde 3P Lipid synthesis Glycolysis 1,3 Biphosphoglycerate 3P Glycerate Amino acid synthesis 2P Glycerate Phosphoenolpyruvate TCA cycle Mitochondrie NAD + NADH, H + IV III II I V ETC 2x ATP Lactate Pyruvate 32x ATP

Increased glucose uptake for detection of immunological disease Glycolytic switch of Hematopoietic cells by FDG positron emission tomography Hodgkin’s Lymphoma Systemic Lupus Erythematosus Pagano et al. Radiology Case Reports 2012 Curiel et al. Ann NY Acad Sci 2011

Glycolysis drives transcriptional programs Glucose ATP Citrate Lyase Glycolysis Mitochondrie Pyruvate TCA citrate Acetyl CoA High glycolytic rates required for eg: transcription of inflammatory cytokines in T cells maintenance of pluripotency in hematopoietic stem cells Moussaieff et al Cell Metabolism 2015, Zheng et al. Cell 2003

Glycolysis drives post-transcriptional programs GAPDH is a key glycoltyic and RNA-binding enzyme. Upon activation of glycolysis, GAPDH relocalizes away from the RNA of IFNy and IL2, allowing translation of these cytokines. Chang et al. Cell 2013; Millet P et al. J Immunol. 2016, White MR et al. RNA 2016

Glycolytic intermediates as signaling molecules Phosphoenolpyruvate modulates SERCA associated Calcium signaling and induces nuclear translocation of NFAT1 Glucose Pyruvate Fructose 1,6biP Fructose 6P Phosphoenolpyruvate 2P Glycerate 3P Glycerate 1,3 Biphosphoglycerate Glyceraldehyde 3P Glucose 6P PCK1-overexpressing T cells restrict tumor growth /prolong survival of melanoma-bearing mice Ho et al. Cell 2015

Glycolysis as target of immune invasion Local availability of glucose is important for T cell functions T cells compete with cancer for glucose Chang et al. Cell 2015

Glycolysis as target of immune invasion Low glycolytic rates in T cells activate microRNAs miR-101 and miR-26a, which reduce expression of methyltransferase EZH2 EZH2: stimulates T cell polyfunctional cytokine expression (via Notch) promotes T cell survival via Bcl-2 signaling Ovarian cancer model Zhao et al. Nature Immunology 2016

Systemic metabolic alterations Acute infection: serum concentrations of acetate (short chain fatty acid) ↑ → Increase of cellular acetate pool of memory T cells (ATP citrate lyase) → GAPDH acetylation and activation → effector functions Listeria monocytogenes memory T cell Balmer et al. Immunity 2016, adapted from Hess et al.

The various roles of glycolysis in T cell biology Summary The various roles of glycolysis in T cell biology Anabolism (protein, lipid, nucleotides…) Energy (ATP) Transcription (acetyl coA) Translation (GAPHD) Signaling intermediates (phosphoenolpyruvate) Target of immune evasion (glucose consumption, miRNAS) Integrates systemic metabolism (acetate)

Metabolism of T cell activation shares common features with Cancer metabolism Quiescent cell Proliferating cell Glucose Glucose Lipids Nucleotides NADPH, ATP Fatty acids Glycolysis Pyruvate Pyruvate Lactate Lipids ATP (O2) Lipids ATP (O2) TCA cycle TCA cycle AA, GSH Glutaminolysis Glutamine Glutamine

T cells use glutamine to maintain OxPhos and cellular ATP Stable Isotope Tracer Analysis of effector T cells Glucose Glutamine deprivation NTP NADPH Glycolysis Pyruvate Lactate Acetyl-CoA OAA Citrate Malate TCA cycle Lipids Lipids α-Ketoglutarate T cells increase glutamine metabolism when glucose is limiting « Metabolic plasticity » NH3 Glutamate ATP? Glutaminolysis Glutamine Adapted from RG Jones, McGill

How do T cells detect and respond to changes in nutrient availability? AMPK couples nutrient availability to T cell effector function T cell metabolic adaptation is AMPK-dependent AMPK inhibits protein and lipid synthesis, and in turn activates glutaminolysis and OxPhos AMPK is required to maintain bioenergetics in glucose deprivation Blagih et al Immunity 2015

Glutathione Primes T Cell Metabolism for Inflammation Glutaminolysis: Glutathione Primes T Cell Metabolism for Inflammation Activated T cells: ROS ↑ Glutathione (GSH) is essential for T cell effector functions Lack of activation of mTOR Lack of expression of NFAT and Myc Reduced glycolysis and glutaminolysis Glucose HK2 Glycolysis PKM2 Pyruvate OAA Lipids TCA cycle Citrate α-Ketoglutarate Glutathione Redox balance NH3 Glutamate Glutaminolysis GLS Glutamine Loss of CD4 and CD8 T cells with high intracellular Glutathione in HIV infection. Roederer et al. Int. Immunology, 1991; Mak et al. Immunity 2017

T cells in the tumor microenvironment Tumor cells deplete critical nutrients (glucose, etc) and increase lactate conc..  T cell effector functions ↓  regulatory T cells (Treg) ↑ Kyn, kynurenines; Trp, tryptophan; AHR; Aryl Hydrocarbon Receptor Siska et al. Trends Immunol 2015

PD-1 and CTLA4 blockage increases the metabolic fitness of effector T cells activity of PI3K/Akt/mTor ↑ cMyc expression ↑ Siska et al. Trends Immunol 2015

T-Cell Metabolism is a Key Determinant in T-Cell Fate, Differentiation, and Function Kouidhi et al. Frontiers in Immunol 2016

Metabolic targets in cancer and immunotherapy