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Principal Investigator: JONATHAN D POWELL
Organization: JOHNS HOPKINS UNIVERSITY
Fiscal Year: 2024
Award: $126,657
Funding agency: National Cancer Institute
The metabolic activity of immune cells is tightly linked to their function. We have been developing a flow
cytometry based platform that enables single cell analysis of traditional cell surface markers combined with
intracellular staining for proteins involved in metabolic programming. The intracellular proteins include carnitine
palmitoyltransferase Ia (CPT1a), which is the rate-limiting enzyme in fatty acid oxidation in mitochondria,
proteins involved in glycolysis such as hexokinase II, and the mitochondrial membrane protein voltage-
dependent anion channel (VDAC). Intracellular staining also detects histone methylation markers. While this
approach was originally developed to predict and track responses to immunotherapy for cancer, we have
recently applied this technology to peripheral blood mononuclear cells (PBMC) from COVID-19 patients. By
interrogating immune-metabolic programs we have defined three distinct and novel populations of immune
cells in the PBMC of COVID-19 patients: He3K27me3+VDAC+ T cells; Hexokinase II+ Granulocytic Myeloid
Derived Suppressor Cells; and CPT1a+VDAC+DR- monocytic MDSC. By interrogating PBMC from SARS-
CoV-2 infected patients, we seek to define novel biomarkers in order to predict severity of disease and track
the course of disease and to define novel surrogate markers for testing therapeutic regimens and identifying
novel therapeutic targets. Furthermore, we will interrogate these three novel cell types as a means of better
understanding the nature of protective and pathogenic COVID-19 immune responses. Specifically, RP1 will
leverage these unique findings to better understand the role of these myeloid cells in innate immunity in SARS-
CoV-2 infection, while this project, RP2, will specifically examine the T cell responses. Indeed, the unique
population of H3K27me3+VDAC+ T cells appear to be exquisitely sensitive to apoptosis, which is prevented by
the pan-caspase inhibitor ZVAD as well as the VDAC inhibitor VBIT-4. To understand the impact of this
population on antiviral immunity, we will investigate whether these T cells are specific for SARS-CoV-2 and
suggest destruction or dysfunction of viral antigen-specific T cells disproportionately. To address this, we have
developed a unique and powerful assay, the functional expansion of specific T cells (FEST) assay that
identifies antigen-specific T cell clonotypes using a T cell receptor sequencing (TCRseq)-based platform. By
combining the FEST assay, which was originally developed to track tumor reactive T cells in cancer patients,
with flow cytometry and TCRseq, we will determine if SARS-CoV-2-specific T cells are uniquely detected in T
cell populations with increased levels of metabolic markers. The completion of these studies will yield a novel
assay that will enable us to predict the severity and track the outcome of the disease course, develop a novel
biomarker for the testing of therapeutic regimens, provide fundamental insight into the pathogenesis of immune
dysfunction, and potentially provide insight and a means for developing novel immune-metabolic based drug
regimens.
Terms: <2019 novel corona virus><2019 novel coronavirus><2019-nCoV><Address><Antigenic Determinants><Apoptosis><Apoptosis Pathway><Assay><Automobile Driving><Binding Determinants><Bioassay><Biological Assay><Biological Markers><Blood granulocytic cell><Blood monocyte><COVID infected patient><COVID patient><COVID positive patient><COVID-19><COVID-19 immune response><COVID-19 infected patient><COVID-19 infection><COVID-19 patient><COVID-19 positive patient><COVID-19 virus><COVID-19 virus infection><COVID19 infection><COVID19 patient><COVID19 positive patient><COVID19 virus><CPT 1><CV-19><Cancer Patient><Carnitine Acyltransferase I><Carnitine O-Palmitoyltransferase><Carnitine Palmitoyltransferase><Carnitine Palmitoyltransferase I><Caspase Inhibitor><Cell Body><Cell Function><Cell Physiology><Cell Process><Cell surface><Cells><Cellular Assay><Cellular Function><Cellular Physiology><Cellular Process><Clinical><CoV-2><CoV2><Coronavirus Infectious Disease 2019><Critical Illness><Critically Ill><Disease><Disease Outcome><Disorder><Drugs><Dysfunction><EC 2.7.1.1><Enzyme Gene><Enzymes><Epitopes><Flow Cytofluorometries><Flow Cytofluorometry><Flow Cytometry><Flow Microfluorimetry><Flow Microfluorometry><Functional disorder><Generations><Glycolysis><Granular Leukocytes><Granulocytic cell><HIV-1><HIV-I><HIV1><HK II><HK2 Protein><Hexokinase 2><Hexokinase II><Human Immunodeficiency Virus Type 1><Human immunodeficiency virus 1><Immune><Immune Diseases><Immune Disorders><Immune Dysfunction><Immune System Diseases><Immune System Disorder><Immune System Dysfunction><Immune System and Related Disorders><Immune response><Immunes><Immunity><Immunodeficiency and Immunosuppression Disorders><Immunologic Diseases><Immunological Diseases><Immunological Dysfunction><Immunological System Dysfunction><Immunological response><Inflammasome><Inflammation><Innate Immunity><Intermediary Metabolism><Link><Lymphocytopenia><Lymphopenia><Marrow monocyte><Mediating><Medication><Metabolic><Metabolic Marker><Metabolic Processes><Metabolism><Mitochondria><Mitochondrial Membrane Protein><Mitochondrial Porin><Monitor><Muscle Form Hexokinase><Muscle Hexokinase-2><Myeloid Cells><Myeloid-derived suppressor cells><Native Immunity><Natural Immunity><Nature><Non-Specific Immunity><Nonspecific Immunity><PBMC><Palmitoylcarnitine Transferase><Palmitylcarnitine Acyltransferase><Pathogenesis><Pathogenicity><Patients><Peripheral Blood Mononuclear Cell><Pharmaceutical Preparations><Physiopathology><Population><Programmed Cell Death><Proteins><R-Series Research Projects><R01 Mechanism><R01 Program><Regimen><Research Grants><Research Project Grants><Research Projects><Role><SARS corona virus 2><SARS-CO-V2><SARS-COVID-2><SARS-CoV-2><SARS-CoV-2 immune response><SARS-CoV-2 infected patient><SARS-CoV-2 infection><SARS-CoV-2 patient><SARS-CoV-2 positive patient><SARS-CoV2><SARS-CoV2 infection><SARS-associated corona virus 2><SARS-associated coronavirus 2><SARS-coronavirus-2><SARS-related corona virus 2><SARS-related coronavirus 2><SARSCoV2><Severe Acute Respiratory Coronavirus 2><Severe Acute Respiratory Distress Syndrome CoV 2><Severe Acute Respiratory Distress Syndrome Corona Virus 2><Severe Acute Respiratory Distress Syndrome Coronavirus 2><Severe Acute Respiratory Syndrome CoV 2><Severe Acute Respiratory Syndrome-associated coronavirus 2><Severe Acute Respiratory Syndrome-related coronavirus 2><Severe acute respiratory syndrome associated corona virus 2><Severe acute respiratory syndrome coronavirus 2><Severe acute respiratory syndrome coronavirus 2 immune response><Severe acute respiratory syndrome coronavirus 2 infection><Severe acute respiratory syndrome related corona virus 2><Severities><Severity of illness><Staining method><Stains><Subcellular Process><Surrogate Markers><T cell receptor repertoire sequencing><T cell receptor sequencing><T cell response><T-Cell Subsets><T-Cells><T-Lymphocyte><T-Lymphocyte Subsets><TCR repertoire sequencing><TCR sequencing><TCR-seq><TCRseq><Technology><Teff cell><Testing><Therapeutic Agents><Treatment Efficacy><Treatment Protocols><Treatment Regimen><Treatment Schedule><Type II Hexokinase><VDAC mitochondrial porin><VDAC proteins><Variant><Variation><Viral Antigens><Virus><Virus Replication><Voltage-Dependent Anion Channel><Wuhan coronavirus><anion-selective channels voltage-dependent><anti-cancer immunotherapy><anti-viral immunity><anticancer immunotherapy><antigen-specific T cells><antiviral immunity><bio-markers><biologic marker><biomarker><cancer immunotherapy><carnitine palmitoyltransferase 1><cell assay><cell type><coronavirus disease 2019><coronavirus disease 2019 immune response><coronavirus disease 2019 infected patient><coronavirus disease 2019 infection><coronavirus disease 2019 patient><coronavirus disease 2019 positive patient><coronavirus disease 2019 virus><coronavirus disease infected patient><coronavirus disease patient><coronavirus disease positive patient><coronavirus disease-19><coronavirus disease-19 patient><coronavirus disease-19 virus><coronavirus infectious disease-19><coronavirus patient><disease severity><driving><drug/agent><effector T cell><fatty acid oxidation><flow cytophotometry><granulocyte><hCoV19><histone methylation><host response><immune system response><immune-based cancer therapies><immunoresponse><immunosuppressive myeloid cells><immunotherapy for cancer><immunotherapy of cancer><infected with COVID-19><infected with COVID19><infected with SARS-CoV-2><infected with SARS-CoV2><infected with coronavirus disease 2019><infected with severe acute respiratory syndrome coronavirus 2><inhibitor><innovate><innovation><innovative><insight><intervention efficacy><life-threatening COVID><life-threatening COVID-19><life-threatening SARS-CoV-2><life-threatening coronavirus disease><life-threatening coronavirus disease 2019><life-threatening severe acute respiratory syndrome coronavirus 2><methylation biomarker><methylation marker><mitochondria porins><mitochondrial><mitochondrial pore protein><monocyte><myeloid suppressor cells><myeloid-derived suppressive cells><nCoV2><new drug target><new drug treatments><new druggable target><new drugs><new marker><new pharmacological therapeutic><new pharmacotherapy target><new therapeutic target><new therapeutics><new therapy><new therapy target><next generation therapeutics><novel><novel biomarker><novel drug target><novel drug treatments><novel druggable target><novel drugs><novel marker><novel pharmaco-therapeutic><novel pharmacological therapeutic><novel pharmacotherapy target><novel therapeutic target><novel therapeutics><novel therapy><novel therapy target><pathophysiology><patient infected with COVID><patient infected with COVID-19><patient infected with SARS-CoV-2><patient infected with coronavirus disease><patient infected with coronavirus disease 2019><patient infected with severe acute respiratory syndrome coronavirus 2><patient with COVID><patient with COVID-19><patient with COVID19><patient with SARS-CoV-2><patient with coronavirus disease><patient with coronavirus disease 2019><patient with severe acute respiratory distress syndrome coronavirus 2><pore forming protein VDAC><prevent><preventing><programs><rational design><response><serious COVID><serious COVID-19><serious SARS-CoV-2><serious coronavirus disease><serious coronavirus disease 2019><serious severe acute respiratory syndrome coronavirus 2><severe COVID><severe COVID-19><severe COVID19><severe SARS-CoV-2><severe acute respiratory syndrome coronavirus 2 infected patient><severe acute respiratory syndrome coronavirus 2 patient><severe acute respiratory syndrome coronavirus 2 positive patient><severe coronavirus disease><severe coronavirus disease 19><severe coronavirus disease 2019><severe severe acute respiratory syndrome coronavirus 2><single cell analysis><social role><suppressive myeloid cells><surrogate bio-markers><surrogate biomarkers><therapeutic efficacy><therapeutic evaluation><therapeutic testing><therapy efficacy><thymus derived lymphocyte><tumor><viral multiplication><viral replication><virus antigen><virus multiplication><voltage-dependent, anion-selective, channel forming protein, mitochondrial>