Elucidating Drug Interactomes for Cardiac Protection by SGLT2 Inhibitors

PROJECT SUMMARY First designed to treat diabetes, sodium-glucose cotransporter-2 inhibitors (SGLT2i) were used to prevent glucose reabsorption in the kidney. Recent clinical trials of SGLT2i further demonstrated an unexpected and substantial reduction in heart failure hospitalizations in patients with and without diabetes. Since SGLT2 is lowly expressed in the heart, its off-target mechanisms present a fascinating opportunity to elucidate cardiac protective targets beyond glycemic control. This K99/R00 application describes a five-year research training plan that leverages (i) human induced pluripotent stem cell-derived cardiovascular cells, (ii) single-cell RNA transcriptomics (scRNA-seq), (iii) metabolomics, (iv) large-scale drug-protein interaction determination, and (v) cell and animal validation to elucidate cardiac-protective mechanisms in health and disease. Given the well- established, off-target protective mechanisms of SGLT2i in the heart, the applicant, Dr. Arianne Caudal, will test the hypothesis that SGLT2i promotes mitochondrial biogenesis and metabolic remodeling, maintaining energy homeostasis in heart failure. In Aim 1 (K99), Dr. Caudal will use a “cell village” multi-omic population screening platform to determine the transcriptomic and metabolomic response conferred by SGLT2i in cardiomyocytes (iPSC-CMs), fibroblasts (iPSC-CFs), and endothelial cells (iPSC-ECs). In Aim 2 (K99), Dr. Caudal will determine the direct protein binding partners of SGLT2i using a cutting-edge proteomics approach in three-dimensional iPSC-derived engineered heart tissues (EHTs). In Aim 3 (R00), Dr. Caudal will validate mitochondrial pathways using pharmacological induction of cardiac dysfunction in iPSC-CMs and a mouse model of pressure overload- induced hypertrophy heart failure. Furthermore, these methodological pipelines provide a springboard of applicability to a range of small molecules, metabolites, and peptides, creating a systems biology niche for Dr. Caudal’s independent work. The proposed studies build upon PI Dr. Arianne Caudal’s well-suited prior training in iPSC modeling, proteomics, and mitochondrial metabolism while providing new training opportunities in (i) precision health, (ii) single-cell multi-omics, and (iii) animal modeling. Mentor Dr. Joseph Wu is a pioneer in iPSCs and cardiovascular biology, and co-mentor Dr. Michael Snyder is a leading expert in single-cell multi- omics and precision medicine. Collaborators and advisory committee members Drs. Zoltan Arany (cardiac metabolism, heart failure), Devin Schweppe (protein interactions), Allis Chien (mass spectrometry), and Sarah Heilshorn (tissue bioengineering) provide additional expertise and guidance. In summary, the well-tailored research training plan, exceptional mentoring team, and outstanding environment at Stanford University are anticipated to help propel Dr. Caudal toward her long-term goal of establishing an independent research program at the intersection of cardiovascular metabolism and systems biology.