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Principal Investigator: Christian J Goossen
Organization: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
Fiscal Year: 2024
Award: $48,974
Funding agency: National Heart Lung and Blood Institute
Project Summary
Pulmonary arterial hypertension (PAH) is a rare, devastating disease with the initial pathognomonic instigator
being a combination of genetic and environmental (injurious) factors including hypoxemia and reactive oxygen
species (ROS) signaling. PAH is characterized by aggressive lung endothelial cell (EC)/smooth muscle cell
(SMC) proliferation/migration (remodeling) in balance with apoptosis, senescence and several other factors all
leading to ensuing right ventricle (RV) failure. Hypoxia and EC injury in rodents spawn physiological and cellular
signaling changes resembling hallmark changes observed in human PAH. We postulated that NADPH oxidase
1 (NOX1) and attendant ROS mediate EC proliferation and hemodynamic changes occurring in PAH. Findings
from our lab have shown induction of NOX1 and ROS in tissue from PAH patients and in human pulmonary
artery endothelial cells (HPAECs). Furthermore, the laboratory’s preliminary findings implicate AMPK isoform
alpha-1 (AMPKα1) as a potential lynchpin in the NOX1 pro-proliferative pathway. Mechanistically speaking, we
deduced that redox-sensitive cysteines (Cys 299 & 304) in AMPKα1 would be susceptible to oxidation, which
would, in turn, augment AMPKα1 activity. Using the bioinformatic tools of STRING, TRANSFAC and Orange, I
discovered that phosphorylated/activated AMPKα1 is associated with cyclin-dependent kinase inhibitor 1
(p21cip1), a member of the Cip/Kip family of cyclin-dependent kinase (CDK) inhibitors. Importantly, the subcellular
localization of p21cip1 dictates its cellular function. In the nucleus, the Cip/Kip family promotes apoptosis and
senescence through retinoblastoma (Rb)-mediated suppression of the cell cycle. In contrast, when
phosphorylated and largely localized in the cytosol, p21cip1 purportedly indirectly actuates (disinhibits) cyclin D
and CDK4/6 leading to Rb inhibition (by phosphorylation) and a permissive cell cycle progression. Therefore, we
postulate that NOX1 mediates oxidative activation of AMPKα1 subunit via oxidizing cysteines 229 and/or 304
(Cys299/304). This is expected to phosphorylate p21cip1 leading to disinhibited binding from cyclin/CDK and cell
cycle progression and EC hyperproliferation in PAH. To test this overarching hypothesis, the following aims will
be tested: (1) To determine whether Nox1-induced oxidation of AMPKα1 elicits p21cip1 family disinhibition of
CyclinD/CDK4/6 and EC proliferation/migration; (2) To Interrogate the effect of EC NOX1 in the promotion of
vascular remodeling and hemodynamic changes in PAH animal models (Mouse and Rat Su/Hx). This proposal
is expected to reveal previously unidentified pathways controlled by NOX1 that participate in the pathology of
PAH. Additionally, an F31 award at this time will enhance my ability to grow as a young scientist and develop
the skills needed to obtain my PhD and pursue a career in the biomedical sciences.
Terms: <Actins><Active Oxygen><Animal Model><Animal Models and Related Studies><Apoptosis><Apoptosis Pathway><Apoptotic><Award><Binding><Bio-Informatics><Bioinformatics><Body Tissues><CCND1 Protein><CD31><CDK Inhibitor Protein><CDK inhibitor p27><CDK2><CDK2 gene><CDK4><CDK4 gene><CDKI Protein><CDKN1B protein><CDKN4 protein><Cancers><Cardiac artery><Causality><Cause of Death><Cell Body><Cell Communication and Signaling><Cell Cycle><Cell Cycle Progression><Cell Division Cycle><Cell Division Kinase 2><Cell Division Kinase 4><Cell Function><Cell Growth in Number><Cell Isolation><Cell Multiplication><Cell Nucleus><Cell Physiology><Cell Process><Cell Proliferation><Cell Segregation><Cell Separation><Cell Separation Technology><Cell Signaling><Cells><Cellular Function><Cellular Physiology><Cellular Process><Cellular Proliferation><Cellular injury><Cessation of life><Common Rat Strains><Confocal Microscopy><Coronary artery><Cyclin D1><Cyclin Gene><Cyclin Kinase Inhibitor><Cyclin-Dependent Kinase 2><Cyclin-Dependent Kinase 4><Cyclin-Dependent Kinase Inhibitor><Cyclin-Dependent Kinase Inhibitor p27><Cyclin-Dependent Kinases><Cyclin-Dependent Protein Kinases><Cyclins><Cysteine><Cytosol><Death><Development><Diagnosis><Disease><Disinhibition><Disorder><Doctor of Philosophy><Endothelial Cells><Endothelium><Enzyme Gene><Enzymes><Equilibrium><Etiology><Exposure to><FLK1><Family><Financial Support><Fostering><G1/S-Specific Cyclin D1><Genetic><Goals><Half-Cystine><Heart><Heart artery><Human><Hypertrophy><Hypoxemia><Hypoxia><Hypoxic><Immunoblotting><In Vitro><Intracellular Communication and Signaling><Involuntary Muscle><Isoforms><KDR gene><Kinases><Kip1 protein><L-Cysteine><Laboratories><Leiomyocyte><Lesion><Literature><Lung><Lung Respiratory System><Malignant Neoplasms><Malignant Tumor><Mass Photometry/Spectrum Analysis><Mass Spectrometry><Mass Spectroscopy><Mass Spectrum><Mass Spectrum Analyses><Mass Spectrum Analysis><Measurement><Mediating><Mediator><Mentorship><Mice><Mice Mammals><Modeling><Modern Man><Molecular Interaction><Murine><Mus><NADPH Oxidase 1><Nucleus><Oncogenesis><Oranges><Oxidation-Reduction><Oxygen Deficiency><Oxygen Radicals><P105-RB><PECAM1><PECAM1 gene><PP110><PRAD1 Protein><PSK-J3><Pathway interactions><Patients><Ph.D.><PhD><Phenotype><Phosphorylation><Phosphotransferase Gene><Phosphotransferases><Physiologic><Physiological><Predisposition><Pro-Oxidants><Programmed Cell Death><Proliferating><Protein Isoforms><Protein Phosphorylation><Proto-Oncogene Proteins c-bcl-1><Pulmonary Artery><Pulmonary Pathology><Pulmonary Vascular Resistance><Pulmonary artery structure><Rat><Rats Mammals><Rattus><Rb Gene Product><Rb Protein><Rb1 Gene Product><Reactive Oxygen Species><Redox><Reporting><Research><Resistance><Retinal Neuroblastoma><Retinoblastoma><Retinoblastoma Associated Protein><Retinoblastoma Protein><Rodent><Rodentia><Rodents Mammals><Role><Science><Scientist><Short interfering RNA><Signal Transduction><Signal Transduction Systems><Signaling><Small Interfering RNA><Smooth Muscle><Smooth Muscle Cells><Smooth Muscle Myocytes><Smooth Muscle Tissue Cell><Staining method><Stains><Subcellular Process><Survival Rate><Susceptibility><Testing><Time><Tissues><Training><Transphosphorylases><VEGF Receptors><VEGFR><VEGFR-2><VEGFR2><VPF Receptor><Vascular Endothelial Cell Growth Factor Receptor><Vascular Endothelial Growth Factor Receptor 2><Vascular Permeability Factor Receptor><Vascular remodeling><Western Blotting><Western Immunoblotting><Wild Type Mouse><Work><antagonism><antagonist><arterial lesion><balance><balance function><bcl-1 Proto-Oncogene Products><bcl-1 Proto-Oncogene Proteins><bcl1 Proto-Oncogene Proteins><bio-informatics tool><bioinformatics tool><biological signal transduction><c-bcl-1 Proteins><career><causation><cdk Proteins><cell damage><cell injury><cell sorting><cellular damage><cyclin D><cyclin-dependent kinase inhibitor 1B><damage to cells><developmental><disease causation><financial assistance><hemodynamics><hypoxemic><hypoxia-induced pulmonary hypertension><hypoxic pulmonary hypertension><immunocytochemistry><in vivo><indexing><inhibitor><injury to cells><interest><lung microvascular endothelial cells><lung pathology><lung vascular endothelial cells><malignancy><member><migration><model of animal><neoplasm/cancer><neoplastic><normoxia><novel><oxidation><oxidation reduction reaction><p27 Kip1 protein><p27 protein><p27-Kip1><p27Kip1 protein><pRB><pathway><permissiveness><programs><protein blotting><pulmonary><pulmonary arterial endothelial cell><pulmonary arterial hypertension><pulmonary artery endothelial cell><pulmonary artery hypertension><pulmonary microvascular endothelial cells><pulmonary vascular endothelial cells><resistant><response><retina neuroblastoma><right heart failure><right sided heart failure><right ventricle failure><right ventricular failure><right ventricular heart failure><senescence><senescent><siRNA><skills><social role><stem><tumorigenesis><wildtype mouse>