This proposal describes a five year career development plan focused on providing the principle investigator (PI) a strong foundation on which to build a successful academic research program. The long-term goal of the PI is to lead a strong NIH funded research program focused on the investigation of the cellular and molecular dysfunction of cardiomyopathy. In the short term, this proposal will allow the PI to develop independent lines of research in dystrophic cardiomyopathy which is both scientifically important and a clear change in direction from her current mentor. The principle investigator has completed a structured fellowship training project at the Medical College of Wisconsin and is currently expanding her scientific skills through a unique integration of interdepartmental resources. This program will promote the command of mitochondrial physiology and induced pluripotent stem cells (iPSCs) as applied to dystrophic cardiomyopathy. The mentor, Dr. Zeljko Bosnjak is an internationally recognized expert on anesthetic preconditioning and mitochondrial function. He is the vice chairman for research in the Department of Anesthesiology and has a long track record of successful post-doctoral fellows and graduate studies, many of which have gone on to successful careers in academia. In addition to the skills and mentoring of Dr. Bosnjak, the PI has cultivated a strong network of collaborators and advisors to provide scientific and career advice. The research will focus on how various dystrophin mutations affect nitric oxide synthase localization and signaling along with mitochondrial function using human cardiomyocytes derived from iPSCs that contain the genetic background of Duchenne and Becker Muscular Dystrophy patients (MD-iPSCs). iPSCs generated from healthy volunteers serve as a control.
The specific aims are to: 1) determine how specific dystrophin mutations increase MD-iPSC-derived cardiomyocytes sensitivity to stress-induced mitochondrial cell death pathways 2) examine the role of nitric oxide synthase in MD-iPSC-derived cardiomyocytes containing different dystrophin mutations and 3) determine whether up-regulating the nitric oxide-cyclic GMP pathway rescues MD- iPSCs from oxidative stress and cell death. This work represents the first human disease model of dystrophic cardiomyopathy. The Medical College of Wisconsin provides an excellent environment of basic cardiovascular research. The Cardiovascular Center and Department of Anesthesiology provides excellent core resources and a strong collaborative environment. In short, this environment provides all of the resources required for the PI to develop strong independent lines of investigation.
Duchenne muscular dystrophy is the most common fatal genetic disease and heart failure is an emerging threat to these patients. This project will lead t an understanding of how the dystrophin gene mutation affects human cardiomyocyte function and will give insights into improving treatment options for muscular dystrophy patients.
|Afzal, Muhammad Z; Reiter, Melanie; Gastonguay, Courtney et al. (2016) Nicorandil, a Nitric Oxide Donor and ATP-Sensitive Potassium Channel Opener, Protects Against Dystrophin-Deficient Cardiomyopathy. J Cardiovasc Pharmacol Ther 21:549-562|
|Cossette, Stephanie M; Bhute, Vijesh J; Bao, Xiaoping et al. (2016) Sucrose Nonfermenting-Related Kinase Enzyme-Mediated Rho-Associated Kinase Signaling is Responsible for Cardiac Function. Circ Cardiovasc Genet 9:474-486|
|Raphael, Roseanne; Purushotham, Diana; Gastonguay, Courtney et al. (2016) Combining patient proteomics and in vitro cardiomyocyte phenotype testing to identify potential mediators of heart failure with preserved ejection fraction. J Transl Med 14:18|
|Afzal, Muhammad Zeeshan; Gartz, Melanie; Klyachko, Ekaterina A et al. (2016) Generation of human iPSCs from urine derived cells of a patient with a novel homozygous PAI-1 mutation. Stem Cell Res 17:657-660|
|Strande, Jennifer L (2015) Haploinsufficiency MYBPC3 mutations: another stress induced cardiomyopathy? Let's take a look! J Mol Cell Cardiol 79:284-6|
|Afzal, Muhammad Z; Strande, Jennifer L (2015) Generation of induced pluripotent stem cells from muscular dystrophy patients: efficient integration-free reprogramming of urine derived cells. J Vis Exp :52032|