Isabel Kaufman

Investigating the Role of Piezo1 in Differentiation of Cardiomyocytes on Varying Hydrogel Stiffnesses

Heart disease is the leading cause of death in the U.S. In this disease, the heart hypertrophies and cardiomyocytes become enlarged. Cardiomyocytes are a non-renewable cell type, so we can’t use primary human cardiomyocytes. Instead, we use induced pluripotent stem cells (iPSCs): somatic cells reprogrammed to become pluripotent, or able to become any cell type. By studying the differentiation process of iPSCs into cardiomyocytes (IPSC-CMs), we can optimize differentiation procedures for research and regenerative medicine. We will also study mechanosensitive channels that may play a role in hypertrophy. Hypertrophy was prevented by knock-out of Piezo1 in mice (Yu et al., 2022). Piezo1 is a mechanosensitive ion channel that opens in response to force. We seek to find how differentiation of iPSCs to iPSC-CMs are impacted by placement on hydrogels of soft and rigid stiffnesses, mimicking healthy and diseased heart tissue, and activation/knockout of Piezo1. We are using a Piezo1 tagged line, a Piezo1 knock-out (KO) line, and a positive-control with Yoda1 (an activator of Piezo1). We predict that Piezo1 KO and differentiation on stiffer gels will show reduced differentiation effectiveness, confluency, and differing morphology. We differentiated 12 plates of iPSCs from each line into cardiomyocytes while imaging plates daily. Using ImageJ, we will gather morphology data (confluency, circularity, perimeter, solidity, etc.). For differentiation effectiveness, we will use another software to count how many cells are beating to determine successful cardiomyocyte differentiation. Our ultimate research goal is to study the effect of Piezo1 on hypertrophy in heart disease.