PROJECT SUMMARY Summary of Parent R01: Type 2 diabetes (T2D) is a long-term metabolic disorder affecting 12% of the US population. It is a leading cause of death nationwide, primarily due to associated cardiovascular disease (CVD, >65% of patients). While CVD risk factors include high cholesterol, hypertension, and smoking, a subset of patients suffer from myocardial dysfunction, a term named type 2 diabetic cardiomyopathy (T2DCM), which suggest factors within the cardiac myocyte itself may give rise to detrimental cardiac remodeling associated with diabetes. Despite the obvious importance of T2DCM, there is currently no specific effective treatment for it and a deep understanding of this complex disease at the molecular level is lacking. Hence, resolving the contributing mechanisms of T2DCM is a pressing goal of basic and translational research. The recent advent of new technological breakthroughs, such as patient-specific human induced pluripotent stem cells (iPSCs) and genome editing, provides an unprecedented opportunity to study associations between genetic variability and disease susceptibility. The overarching goal of our multi-PI R01 grant is to understand the underlying mechanisms of T2DCM using patient-specific iPSC-derived cardiomyocytes and endothelial cells from T2D patients and to understand individual susceptibility to disease development. We have assembled a team of highly accomplished clinicians and researchers in cardiac stem cell biology, genomics, molecular genetics, biostatistics and bioinformatics. We are well positioned to achieve the project goals within five years. Our proposal will enable a novel personalized approach to better understand the mechanisms underlying T2DCM that could ultimately revolutionize treatment strategies. Proposed Supplement: Hispanics and non-Hispanic Asians account for 23% of the US population based on recent CDC data released in December 2019 and from a recent study published in the Journal of the American Medical Association. Among 2,266 individuals diagnosed with diabetes of 7,575 US adults, the prevalence of diabetes in the Hispanic population was 22.1%. In the proposed diversity supplement, we will extend the scope of parent R01 to include an additional patient demographic involving Hispanics. Specifically, we will expand the patient cohort with 20 additional individuals (10 T2DCM, 10 Controls) from the Hispanic- American community. We will generate 3D cardiac organoids to characterize the functional changes in cardiomyocytes and endothelial cells. These cardiac organoids will be exposed to hyperglycemic and normoglycemic conditions to unravel the transcriptomic landscape using single-cell RNA sequencing. The experiments will be carried out by Ms. Nicole Lopez.

PROJECT NARRATIVE Diabetic cardiomyopathy is associated with other risk factors such as hypertension and coronary artery disease, it is well known that chronic type-2 diabetes impairs heart function independent of these risk factors. Induced pluripotent stem cell (iPSCs) derived from diabetic patients is an indispensable tool to study pathological signatures that lead to metabolic disorder affecting multiple organ systems. The parent R01 is focused on elucidating the molecular and cellular basis and genotype-phenotype correlation of type-2 diabetes induced cardiomyopathy (T2DCM). For this R01 diversity supplement, we aim to study the dysfunction in using 3-D cardiac organoids generated from Hispanic Americans who are at a higher risk of developing T2DCM in the United States.