Project Summary/Abstract: This proposal outlines a 5-year mentored career development plan as a physician-scientist for Anita Saraf, MD, PhD as the principal investigator. Dr. Saraf is a tenure-track Assistant Professor at University of Pittsburgh School of Medicine. She completed her graduate training through the Medical Scientist Training Program at Baylor College of Medicine, Houston, TX with a Ph.D. in Bioengineering through the William Marsh Rice University. Subsequently, she completed her residency and fellowship through the ABIM research pathway at Emory University, Atlanta, GA and completed subspeciality training in Adult Congenital Heart Disease. Dr. Saraf’s post-graduate research has focused on translational and basic science studies involving biomarker profiles in single ventricle Fontan patients and understanding the effect of these biomarkers in cardiomyocytes derived from induced pluripotent stem cells (iPSCs). Under the combined mentorship of Drs. Bernhard Kühn and Toren Finkel (co-mentor), Dr. Saraf proposes to further her research skill-set in gene-editing and cardiomyocyte physiology, with a long-term goal of establishing herself as a leader in translational and basic science research in congenital heart disease. In this proposal, Dr. Saraf investigates the role of NOTCH1 hypomorphic mutations in inducing arrhythmias and heart failure in patient-derived and engineered cardiomyocytes from iPSCs. Using iPSCs from NOTCH1 hypomorphic patients with hypoplastic left heart syndrome (HLHS) and engineering similar hypomorphic mutations in control iPSCs with CRISPR gene-editing, Dr. Saraf proposes the following objectives: (1) Determine the influence of inflammatory cytokines (found to chronically elevated in univentricular patients) on hypomorphic NOTCH1 cardiomyocytes and (2) Determine the influence of systemic pressures on hypomorphic NOTCH1 cardiomyocytes, with respect to abnormal calcium-handling and contractility. Given the lack of viable animal models for congenital heart disease, iPSC-derived cardiomyocytes together with gene-editing technology provides a powerful platform to understand abnormalities in cardiomyocyte physiology, discover novel targets for therapy, and create viable animal models with similar hypomorphic mutations in the future. In addition to research training, Drs. Kühn, Finkel and Saraf have formulated a clear timeline for career development involving publications, presentations at meetings and courses in leadership that will transition Dr. Saraf to independence as a leader in this field.