This proposal outlines a comprehensive 4-year training program to develop Veli K. Topkara, MD, MSc, an Assistant Professor of Medicine at Columbia University, into an independent translational investigator. Dr. Topkara is an advanced heart failure (HF) cardiologist whose long-term goal is to understand basic mechanisms responsible for maladaptive cardiac remodeling with an emphasis on long non-coding RNAs (lncRNAs), in order to improve outcomes of patients with heart failure. The career development plan and mentorship structure proposed in this application are designed to fill specific educational and experiential gaps in Dr. Topkara?s training, ensuring his maturation into an independent physician-scientist. His short-term career goals include: 1) to develop skills in design and conduct of mechanistic translational research with a specific focus on lncRNA biology, 2) to acquire expertise in generation of induced-pluripotent stem cell derived cardiomyocytes (iPS-CMs) and gene editing technology, 3) to develop experience in computational and bioinformatics analysis of lncRNAs, 4) acquire skills in development and application of in vivo lncRNA knockdown strategies, and 5) to develop grant- writing skills. The mentorship team represents a multidisciplinary group of individuals specifically chosen to achieve PI's particular scientific and career development goals. At an institutional level, Columbia provides an incredibly rich and supportive intellectual and collaborative research environment with ample resources available for Dr. Topkara to successfully accomplish his research and career development goals. Through RNA sequencing of failing human heart samples, Dr. Topkara identified 81 candidate lncRNAs that are significantly dysregulated in HF. Based on (i) cardiac tissue enrichment in Genotype-Tissue Expression (GTEx) dataset, (ii) reversibility with left ventricular assist device support, (iii) locus conservation in mouse, (iv) modulation during cardiomyocyte hypertrophy; lnc-TECRL (lnc-Trans-2,3-Enoyl-CoA Reductase Like) was selected for structural and functional characterization. Based on preliminary data, the main hypothesis of this proposal is that lnc- TECRL mediates pathological cardiac hypertrophy through its interaction with chromatin modifying proteins and activation of the hypertrophic gene program.
The specific aims are to: 1) Determine mechanisms by which lnc- TECRL mediates cardiomyocyte hypertrophy in vitro and 2) Investigate the therapeutic potential of lnc-TECRL in prevention of maladaptive cardiac remodeling in vivo. To achieve these goals, the PI will utilize adult human cardiomyocytes, human iPS-CMs, mature engineered human cardiac tissues, and mouse models which will establish functional and therapeutic relevance of lnc-TECRL in cardiac remodeling. To understand mechanisms by which lnc-TECRL mediates cardiac hypertrophy, state-of-art techniques including RNA pull-down, RNA-IP, CHIP, domain mapping by deletion, ChiRP-seq, and in vivo GapmeR knockdown will be employed. Completion of the proposed work is expected to provide novel insights into the transcriptional regulation of maladaptive cardiac remodeling and uncover new approaches that could be translated for therapeutic benefit in HF patients. .
Long non-coding RNAs (lncRNAs) are emerging as causal in complex disease states, but the roles of lncRNAs in heart failure are largely unknown. In this proposal, the PI will perform mechanistic interrogation of lnc-TECRL in modulating pathological cardiac hypertrophy and remodeling. The research outlined in this proposal has broad novel significance and will seed innovative translation of lncRNAs for novel diagnostic and therapeutic targeting in human heart failure.