DESCRIPTION (provided by applicant): The central theme of our PPG is to gain fundamental insight into the mechanisms responsible for greater susceptibility of diabetic hearts to ischemia-reperfusion injury using both patient-derived cardiomyocytes and diabetic animals. We will use anesthetics to test the ability of diabetic animals and human cardiomyocytes to be protected against ischemia-reperfusion injury for the future development of novel cardioprotective strategies for the diabetic heart. Our hypothesis is that diabetes undermines cardioprotection through actions on cardiomyocytes that are both environmental and cellular in origin. Our translational approach will include in vivo diabetic animal models, isolated animal heart preparations, isolated animal cardiomyocytes and mitochondria, computer simulations and modeling of mitochondrial and cellular function in anesthetic cardioprotection, and finally, human cardiomyocytes derived from disease-specific induced pluripotent stem cells (iPSC). The following closely interrelated and interdependent Projects will address different facets of this theme: Project I (PI: Zeljko J. Bosnjak - Anesthesiology) will utilize human cardiomyocytes derived from the iPSC obtained from non-diabetic individuals and patients with type 2 diabetes mellitus along with a rat model of type 2 diabetes developed at the Medical College of Wisconsin (T2DNmtFHH or T2DN for short). Our fundamental hypothesis is that diabetes and glucolipotoxicity impair anesthetic cardioprotection through signaling components that can be favorably modulated to restore anesthetic cardioprotection during diabetes. Project II (PI: Mingyu Liang - Physiology) will examine the role of miR-21 in anesthetic cardioprotection in non-diabetic and diabetic mice and rats, translate the findings to human using patient-specific cardiomyocytes, and investigate the molecular mechanisms involved. This project will test the hypothesis that a change of miR- 21 expression will restore isoflurane-conferred protection in diabetic animal models and in patient-specific cardiomyocytes. Project III (PI: Ranjan K. Dash - Biotechnology and Bioengineering Center and Physiology) will use a system biology approach to iteratively conduct experiments and use the measured data to computationally model and mechanistically characterize the specific effects and associated mechanisms of volatile anesthetic action on mitochondrial and cellular function that lead to cardioprotection, and how diabetic conditions impair this protection. Administrative Core (Director: Zeljko J. Bosnjak - Anesthesiology) will provide an organized and comprehensive framework of support for all subprojects and cores contributing to the cohesive functionality of the Program while ensuring regulatory compliance. Stem Cell Core (Director: Stephen A. Duncan - Cell Biology, Neurobiology and Anatomy) will support all projects by supplying patient-derived iPSC lines and differentiate these cells into disease-specific cardiomyocytes. Thus, all three Projects focus cohesively on the role of diabetes undermining the cardioprotection. This is a highly focused Program that will be led by investigators who have worked together for many years, and their history of collaboration has resulted in closely linked individual Projects that are ideally suitedfor the continuation of this PPG. These investigations will be supported by well-established state-of-the-art facilities in Anesthesiology Research, and the Departments of Physiology; Biophysics; Biochemistry; Pharmacology; Cell Biology, Neurobiology and Anatomy; Medicine; Cardiovascular Research Center; Human and Molecular Genetic Center and Biotechnology and Bioengineering Center.

PUBLIC HEALTH RELEVANCE: The cause of greater cardiac susceptibility to stress in diabetic patients remains unknown. For the first time we are able to make human disease-specific cardiac cells derived from their skin cells. Hence, we can now assess separately the role of cellular and environmental factors that are responsible for greater cardiac sensitivity in patients with diabetes. Defects observed during our study will be targeted with pharmacological therapies in order to restore anesthetic-induced cardioprotection.