Document text
Principal Investigator: SACHIN A GUPTE
Organization: NEW YORK MEDICAL COLLEGE
Fiscal Year: 2020
Award: $547,406
Funding agency: National Heart Lung and Blood Institute
Glucose-6-phosphate dehydrogenase (G6PD) is a key enzyme in glucose metabolism and is also a master
regulator of cellular redox signaling that plays a crucial role in the regulation of vascular function. We recently
isolated and characterized long (G6PD545) and short (G6PD515) G6PD isoforms in vascular smooth muscle cells
(VSMCs). Our preliminary data demonstrate that G6PD545 has a putative nuclear localization sequence (NLS)
and is predominantly localized in the nucleus of synthetic VSMCs. We further show that nuclear G6PD545 (but
not cytosolic G6PD515) represses expression of myocardin (MYOCD), the major switch for the VSMC contractile
phenotype, thereby mediating VSMC reversion to a synthetic state, which is considered pathognomonic for a
number of vascular complications, including atherosclerosis, transplant arteriopathy and bypass graft failure.
Importantly, we found that arteries from rats, pigs and humans with metabolic syndrome all exhibit VSMC
synthetic phenotype-associated atherosclerosis and contain high levels of nuclear G6PD545. Interestingly,
individuals harboring a loss-of-function SNP within exon 6 of G6PD (Mediterranean-type G6PD mutation)
exhibit 80% less G6PD activity and are less likely to develop cardiovascular disease. These observations lay the
foundation for our hypothesis that up-regulation of a novel isoform of G6PD functions as a NADPH-
dependent repressor of MYOCD expression that promotes a synthetic VSMC phenotype and pathogenic
vascular remodeling. This novel hypothesis will be tested through a series of interrelated specific aims
designed to elucidate the transcriptional control of MYOCD expression by G6PD545 and the role of G6PD545 in
injury-induced neointimal vascular disease in normal and metabolic syndrome rats. Aim 1 will determine
whether G6PD545 functions as a switch promoting a VSMC synthetic phenotype in balloon-injured rat carotid
arteries in which G6PD545 is down-regulated through Cre/loxP knockdown or CRISPR-Cas9-mediated genome
editing to model the Mediterranean-type G6PD mutation. The effects of G6PD545 ± NLS and G6PD515 vs.
G6PD545 overexpression on VSMC phenotype will also be studied. Aim 2 will determine whether G6PD545 gain-
of-function promotes a VSMC synthetic phenotype through repression of MYOCD expression in cultured
VSMCs as well as vascular tissue from healthy vs. balloon-injured rat carotid arteries. Additionally, the effect
of loss- and gain-of-function of MYOCD on G6PD545-induced changes in VSMC marker genes will be
determined. Aim 3 will elucidate the mechanism of G6PD545 up-regulation and evaluate its role in switching
VSMCs to a synthetic phenotype in models of pathological vascular remodeling (e.g., rat metabolic syndrome).
This project will reveal a heretofore unrecognized relationship between G6PD545 and the regulation of VSMC
phenotype and vascular remodeling in response to physical injury and metabolic disease. The results of these
studies will have enormous applicability for development of new therapies to combat occlusive vascular
diseases and perhaps other diseases in which MYOCD expression may be altered (e.g., asthma, hypertension).
Terms: <2019 novel coronavirus><2019-nCoV><21+ years old><Acute><Adult><Adult Human><Affect><Ailmentary System><Alimentary System><Angiotensin Converting Enzyme><Angiotensin I-Converting Enzyme><Antiviral Agents><Antiviral Drugs><Antivirals><Arterial Disorder><Arteries><Arteriopathy><Asthma><Atheroscleroses><Atherosclerosis><Atherosclerotic Cardiovascular Disease><Blood Vessels><Body Tissues><Brain Stem><Brainstem><Bronchial Asthma><Bypass><CD143 Antigens><COVID-19><COVID19><CRISPR method><CRISPR methodology><CRISPR technique><CRISPR technology><CRISPR-CAS-9><CRISPR-based method><CRISPR-based technique><CRISPR-based technology><CRISPR-based tool><CRISPR/Cas method><CRISPR/Cas technology><CRISPR/Cas9><CRISPR/Cas9 technology><Carboxycathepsin><Cardiovascular><Cardiovascular Body System><Cardiovascular Diseases><Cardiovascular Organ System><Cardiovascular system><Carotid Arteries><Cas nuclease technology><Cell Body><Cell Communication and Signaling><Cell Nucleus><Cell Signaling><Cells><Cellular Membrane><Cessation of life><Cleaved cell><Clustered Regularly Interspaced Short Palindromic Repeats method><Clustered Regularly Interspaced Short Palindromic Repeats methodology><Clustered Regularly Interspaced Short Palindromic Repeats technique><Clustered Regularly Interspaced Short Palindromic Repeats technology><CoV S protein><CoV glycoprotein S><CoV spike glycoprotein><CoV spike protein><Coenzyme II><Common Rat Strains><Coronavirus glycoprotein S><Coronavirus spike protein><Cre-Lox><Cre-LoxP><Cre/LoxP><Data><Death><Development><Diabetes Mellitus><Digestive System><Dipeptidyl Peptidase A><Disease><Disorder><EC 1.1.1.49><Endocytosis><Endothelial Cells><Enzyme Gene><Enzymes><Epithelial Cells><Epitheliasin Gene><Esteroproteases><Exhibits><Exons><Failure><Family member><Family suidae><Foundations><Gastrointestinal Body System><Gastrointestinal Organ System><General Population><General Public><Genes><Genetic Alteration><Genetic Change><Genetic defect><Glucose-6-Phosphate Dehydrogenase><Glucosephosphate Dehydrogenase><Heart Vascular><Human><Human Genome><Hyperplasia><Hyperplastic><Hypertension><Hypertensive Crisis><Individual><Infection><Inflammation><Injury><Intervention><Intervention Strategies><Intracellular Communication and Signaling><Isoforms><Kininase A><Kininase II><Leiomyocyte><Lung><Lung Respiratory System><Lung diseases><Medial><Mediating><Mediator><Mediator of Activation><Mediator of activation protein><Metabolic Diseases><Metabolic Disorder><Metabolic syndrome><Mice><Mice Mammals><Modeling><Modern Man><Morbidity><Morbidity - disease rate><Murine><Mus><Mutation><NAD phosphate><NAD(H) phosphate><NADH phosphate><NADP><NADPH><Nicotinamide-Adenine Dinucleotide Phosphate><Nuclear><Nucleus><Ortholog><Orthologous Gene><Oxidation-Reduction><PRSS10><Pathogenesis><Pathogenicity><Pathologic><Pathologic Constriction><Pathological Constriction><Patients><Peptidases><Peptide Hydrolases><Peptidyl-Dipeptidase A><Phenotype><Pigs><Play><Predisposition><Protease Gene><Proteases><Protein Isoforms><Proteinases><Proteins><Proteolytic Enzymes><Pulmonary Diseases><Pulmonary Disorder><Pulmonary Hypertension><Rat><Rats Mammals><Rattus><Redox><Regulation><Repression><Respiratory Disease><Respiratory System Disease><Respiratory System Disorder><Role><SARS Virus><SARS corona virus><SARS coronavirus><SARS-Associated Coronavirus><SARS-CoV><SARS-CoV-2><SARS-CoV2><SARS-Related Coronavirus><SARS-associated coronavirus 2><SARS-coronavirus-2><SARS-related coronavirus 2><Series><Serine Endopeptidases><Serine Protease><Serine Protein Hydrolases><Serine Proteinases><Severe Acute Respiratory Syndrome Virus><Severe Acute Respiratory Syndrome corona virus><Severe Acute Respiratory Syndrome coronavirus><Severe acute respiratory syndrome coronavirus 2><Severities><Signal Transduction><Signal Transduction Systems><Signaling><Smooth Muscle Cells><Smooth Muscle Myocytes><Smooth Muscle Tissue Cell><Social Distance><Stenosis><Stents><Suidae><Susceptibility><Swine><Syndrome><Systemic hypertension><TMPRSS2><TMPRSS2 gene><Testing><Therapeutic Intervention><Thesaurismosis><Tissues><Transcription Regulation><Transcriptional Control><Transcriptional Regulation><Transmission><Transplantation><Triphosphopyridine Nucleotide><United States><Up-Regulation><Upregulation><Vascular Diseases><Vascular Disorder><Vascular Hypertensive Disease><Vascular Hypertensive Disorder><Vascular Smooth Muscle><Vascular remodeling><Virus><Wuhan coronavirus><adulthood><age group><anti-viral agents><anti-viral drugs><anti-virals><atheromatosis><atherosclerotic disease><atherosclerotic vascular disease><biological signal transduction><blood vessel disorder><cardiovascular disorder><circulatory system><cleaved><co-morbid><co-morbidity><combat><comorbidity><corona virus disease 2019><coronavirus S protein><coronavirus disease 2019><coronavirus spike glycoprotein><cytokine release syndrome><cytokine storm><design><designing><developmental><diabetes><disease of the lung><disorder of the lung><gain of function><gastrointestinal system><genome editing><genome mutation><genomic editing><glucose metabolism><graft failure><high blood pressure><human whole genome><hyperpiesia><hyperpiesis><hypertensive disease><injured><injuries><intervention therapy><interventional strategy><knock-down><knockdown><loss of function><lung disorder><metabolism disorder><mortality><mouse genome><myocardin><new drug treatments><new drugs><new therapeutics><new therapy><next generation therapeutics><novel><novel drug treatments><novel drugs><novel therapeutics><novel therapy><overexpress><overexpression><oxidation reduction reaction><pandemic><pandemic disease><porcine><post-transplant><post-transplantation><posttransplant><posttransplantation><prevent><preventing><protein expression><pulmonary><response><restenosis><severe acute respiratory syndrome-CoV><social role><suid><targeted drug therapy><targeted drug treatments><targeted therapeutic><targeted therapeutic agents><targeted therapy><targeted treatment><transmission process><transplant><vascular><vascular dysfunction><vasculopathy>