Date of Award

Fall 2023

Project Type

Dissertation

Program or Major

Genetics

Degree Name

Doctor of Philosophy

First Advisor

Feixia F.C. Chu

Second Advisor

Feixia Chu

Third Advisor

Xuanmao Chen

Abstract

Heart disease is the leading cause of death in the United States, constituting a quarter of total deaths. A major reason behind the high mortality of heart disease is the low regenerative capacity of cardiomyocytes (CMs), which carry out the contractile function of the heart. Lineage- specific differentiation of pluripotent stem cells into CMs provides a promising therapeutic strategy. However, current methods suffer from challenges in generating mature and homogenous CMs in sufficient quantities for practical application. Moreover, little is known about the underlying epigenetic regulations in CM differentiation, which is essential during the establishment and maintenance of cell identity and lineage specification. To address this knowledge gap, I have compared the histone epigenetic states of embryonic stem cells (ESCs) and CM-enriched cells, as well as ESCs cultured on two-dimensional plates and in three-dimensional hydrogel scaffolds. The most prominent histone modification change upon CM differentiation is identical to that cultured in hydrogel, suggesting a plausible mechanism in hydrogel-promoted CM differentiation. In addition, our phosphoproteomics analysis uncovered the involvement of multiple mechanosensing pathways in pluripotent stem cells responding to three-dimensional hydrogel environment, as well as a putative pathway that least to the change in histone epigenetic states. For high throughput screening of various differentiation conditions, I have generated a stable ESC line with green fluorescence protein with the promoter of a mature cardiomyocyte marker (myosin heavy chain). These results and reagents improve our understanding about CM differentiation, and elucidate promising biological targets in developing an effective CM differentiation method towards cardiac regenerative therapies.

Available for download on Thursday, September 11, 2025

Share

COinS