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Principal Investigator: Jeffrey C Rathmell
Organization: VANDERBILT UNIVERSITY MEDICAL CENTER
Fiscal Year: 2024
Award: $511,194
Funding agency: National Cancer Institute
SUMMARY
Exploiting immunity to eliminate cancer cells offers tremendous new therapeutic opportunities, including the
widely employed immune checkpoint blockade (ICB) agents. These approaches are challenged, however, by
the multitude of mechanisms through which tumors can suppress anti-tumor immunity and render them effective
in only a portion of patients. We have shown that effector T cells require high rates of glucose uptake for anabolic
metabolism, and it is now apparent that cancer cells and the tumor microenvironment (TME) disrupt anti-tumor
immunity in part through metabolic immune suppression. To address this barrier to immunotherapy, we
examined tumor infiltrating lymphocytes (TIL) from surgically excised samples of human clear cell Renal Cell
Carcinoma (ccRCC), a cancer with moderate rates of ICB response that is characterized by loss of the Von
Hippel-Lindau (VHL) tumor suppressor. These tumors allowed us to show that both glucose and glutamine are
available in the TME and while glucose metabolism promotes effector T cells, metabolism of glutamine restrains
T cell effector function. It is unclear which glutamine-dependent enzymes or metabolites suppress T cells, but
we found that Glutaminase-deficiency altered histone methylation and reduced expression of Pik3ip3, a PI3K-
inhibitory protein that suppresses PI3K/mTORC1 signaling and production of inflammatory effector cytokines.
To further explore mechanisms of T cell suppression by glutamine, we performed an in vivo CRISPR screen in
primary TIL and found loss of Glutamine Synthetase among all glutamine-metabolizing enzymes to most
effectively increase TIL accumulation. We also directly defined cell type-specific glucose and glutamine usage
in the TME using radiolabeled Positron Emission Tomography tracers. In contrast to classic Warburg
metabolism, tumor associated macrophages (TAM) were the dominant consumers of glucose, followed by TIL,
and cancer cells, which instead preferentially consumed glutamine. Interestingly, loss of Vhl did not increase
glucose uptake of RCC cells in vivo but instead increased glucose uptake in TIL and TAM. To explore these
pathways in patients undergoing ICB therapy, we next performed high dimensional CyTOF analyses of
peripheral blood from a longitudinal cohort of patients before and 3 weeks after start of therapy. This approach
specifically identified rare but highly proliferative ICB-responsive CD8 and CD4 T cells with elevated
mitochondrial potential. Based on these findings, we hypothesize that RCC genetics drive a metabolically
immunosuppressive TME with abundant glutamine that suppresses PI3K signaling to impair T cell
effector differentiation and function. We will study primary human ccRCC tumors and mouse RCC models to:
(1) Test how nutrients in the ccRCC TME and tumor genetics influence TIL function and metabolism; and (2)
Determine how glucose and glutamine metabolism in the TME promote or suppress anti-tumor immunity.
Together, these studies will establish mechanisms by which glutamine impairs T cell differentiation and test new
potential targets to overcome metabolic immune suppression in the TME to improve anti-tumor immunity.
Terms: <1-Phosphatidylinositol 3-Kinase><3-D><3-Dimensional><3D><Address><Affect><B7-1><BB1><Breast Cell Glutaminase><CD152><CD152 Antigen><CD152 Gene><CD28LG><CD28LG1><CD4 Cells><CD4 Positive T Lymphocytes><CD4 T cells><CD4 helper T cell><CD4 lymphocyte><CD4+ T-Lymphocyte><CD4-Positive Lymphocytes><CD8><CD80><CD80 gene><CD8B><CD8B1><CD8B1 gene><CRISPR editing screen><CRISPR screen><CRISPR-based screen><CRISPR/Cas9 screen><CTLA 4><CTLA-4 Gene><CTLA4><CTLA4 gene><CTLA4-TM><Cancer Cause><Cancer Etiology><Cancer Treatment><Cancers><Cell Body><Cell Communication and Signaling><Cell Function><Cell Physiology><Cell Process><Cell Shape><Cell Signaling><Cells><Cellular Function><Cellular Physiology><Cellular Process><Cellularity><Clear cell renal cell carcinoma><Complex><Consumption><Cytotoxic T-Lymphocyte Protein 4><Cytotoxic T-Lymphocyte-Associated Antigen 4><Cytotoxic T-Lymphocyte-Associated Protein 4><Cytotoxic T-Lymphocyte-Associated Serine Esterase-4><D-Glucose><Dextrose><Dissociation><EC 3.5.1.2><Enzyme Gene><Enzymes><GA Protein><Genes><Genetic><Gln><Glucose><Glutamate Ammonia Ligase (ADP)><Glutamate-Ammonia Ligase><Glutamates><Glutaminase><Glutamine><Glutamine Synthetase><Glycolysis><Human><Hypoxia><Hypoxic><Immune><Immune Cell Activation><Immune mediated therapy><Immune system><Immunes><Immunity><Immunologically Directed Therapy><Immunosuppression><Immunosuppression Effect><Immunosuppressive Effect><Immunotherapy><Impairment><In Vitro><Inflammatory><Intercellular Fluid><Intermediary Metabolism><Interstitial Fluids><Intervention><Intervention Strategies><Intracellular Communication and Signaling><L glutamine amidohydrolase><L-Glutamate><L-Glutamine><LAB7><LYT3><Liver Glutaminase><Long-term cohort><Longitudinal cohort><Longterm cohort><Lymphocyte Function><Malignant Cell><Malignant Neoplasm Therapy><Malignant Neoplasm Treatment><Malignant Neoplasms><Malignant Tumor><Mediating><Mediator><Metabolic><Metabolic Pathway><Metabolic Processes><Metabolism><Mice><Mice Mammals><Mitochondria><Modeling><Modern Man><Murine><Mus><Nutrient><Nutrient availability><Operative Procedures><Operative Surgical Procedures><Organoids><Oxygen Deficiency><PD-1/PD-L1><PD-1/PDL1><PD1-PD-L1><PD1/PD-L1><PD1/PDL1><PET><PET Scan><PET imaging><PETSCAN><PETT><PI-3 Kinase><PI3-Kinase><PI3CG><PI3KGamma><PI3k><PIK3><PIK3CG><PIK3CG gene><Pathway interactions><Patients><Phosphatidylinositol 3-Kinase><Phosphatidylinositol-3-OH Kinase><Phosphoinositide 3-Hydroxykinase><Play><Population><Positron Emission Tomography Medical Imaging><Positron Emission Tomography Scan><Positron-Emission Tomography><Primary Neoplasm><Primary Tumor><Process><Production><Proteins><PtdIns 3-Kinase><Q Levoglutamide><Q. Levoglutamide><Rad.-PET><Radiolabeled><Research Resources><Resources><Role><Sampling><Shapes><Signal Transduction><Signal Transduction Systems><Signaling><Subcellular Process><Surgical><Surgical Interventions><Surgical Procedure><T cell differentiation><T-Cell Activation><T-Cells><T-Lymphocyte><T4 Cells><T4 Lymphocytes><Teff cell><Testing><Tracer><Tumor Cell><Tumor Immunity><Tumor Promotion><Tumor Suppressor Proteins><Tumor Tissue><Tumor-Infiltrating Lymphocytes><Tumor-associated macrophages><Type I Phosphatidylinositol Kinase><Type III Phosphoinositide 3-Kinase><Waste Products><activate T cells><antagonism><antagonist><anti-cancer therapy><anti-tumor immunity><antitumor immunity><biological signal transduction><cancer cell><cancer cell metabolism><cancer immunity><cancer metabolism><cancer microenvironment><cancer therapy><cancer type><cancer-directed therapy><ccRCC><cell type><check point blockade><checkpoint blockade><clustered regularly interspaced short palindromic repeats screen><cytokine><cytotoxic T-lymphocyte antigen 4><effector T cell><glucose metabolism><glucose uptake><glutamate synthetase><glutamatergic><glutamine synthase><high dimensionality><histone methylation><immune activation><immune check point blockade><immune checkpoint blockade><immune microenvironment><immune suppression><immune suppressive activity><immune suppressive function><immune therapeutic approach><immune therapeutic interventions><immune therapeutic regimens><immune therapeutic strategy><immune therapy><immune-based therapies><immune-based treatments><immuno therapy><immunosuppressive activity><immunosuppressive function><immunosuppressive microenvironment><immunosuppressive response><immunosuppressive tumor microenvironment><improved><in vivo><interventional strategy><kidney cortex><kidney cortical portion><malignancy><mitochondrial><neoplasm/cancer><neoplastic cell><new drug treatments><new drugs><new pharmacological therapeutic><new therapeutics><new therapy><next generation therapeutics><novel drug treatments><novel drugs><novel pharmaco-therapeutic><novel pharmacological therapeutic><novel therapeutics><novel therapy><pathway><peripheral blood><positron emission tomographic (PET) imaging><positron emission tomographic imaging><positron emitting tomography><radiolabeling><radiologically labeled><renal cortex><response><restraint><social role><standard of care><surgery><targeted drug therapy><targeted drug treatments><targeted therapeutic><targeted therapeutic agents><targeted therapy><targeted treatment><three dimensional><thymus derived lymphocyte><tumor><tumor cell metabolism><tumor immune microenvironment><tumor metabolism><tumor microenvironment><tumor suppressor><tumor-immune system interactions><uptake>