Date of Award

5-10-2026

Date Published

June 2026

Degree Type

Dissertation

Degree Name

Doctor of Philosophy (PhD)

Department

Biomedical and Chemical Engineering

Advisor(s)

Zhen Ma

Keywords

Cardiotoxicity;costamere;high-content imaging;hiPSC-CM;Machine learning

Abstract

Cardiotoxicity remains a major cause of drug attrition in preclinical development and clinical use, underscoring the need for predictive, mechanistically informative in vitro platforms. Human-induced pluripotent stem cell–derived cardiomyocytes (hiPSC-CMs) provide a physiologically relevant system, yet conventional assays often focus on single readouts and fail to capture the interplay between electrophysiological, mechanical, and structural domains. To address these limitations, this dissertation establishes an integrated physiomics framework combining high-content imaging, genetic editing, traction force microscopy, and computational analysis to characterize cardiomyocyte function at the single-cell level. By simultaneously monitoring action potential dynamics, sarcomeric movement, contractile motion, and mechanical output, the system minimizes batch variation and generates multidimensional datasets capturing overt and subtle drug-induced perturbations. Machine learning uncovers latent phenotypic patterns and improved predictive power: unsupervised analyses revealed drug-specific signatures, while supervised models accurately classified compounds into mechanistic categories and anticipated toxic liabilities. This integrative approach enhances mechanistic interpretation and predictive accuracy in cardiotoxicity assessment. Costameres, as striated muscle–specific cell adhesions, anchor M-lines and Z-lines of the sarcomeres to the extracellular matrix. Previous studies have shown that costameres participate in the initial assembly of myofibrils. However, how costamere maturation cooperates with myofibril growth remains underexplored. In this work, we analyzed zyxin (costameres), α-actinin (Z-lines), and myomesin (M-lines) to track the behaviors of costameres and myofibrils in hiPSC-CMs. We quantified costamere assembly and maturation during myofibril growth in a time-dependent manner. We found asynchrony not only between myofibril and costamere maturation, but also between the formation of Z-costameres and M-costameres associated with distinct sarcomeric structures. This study provides insight into how costameres assemble and integrate with cardiomyocyte sarcomeres, shedding light on cardiomyocyte mechanobiology.

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Open Access

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Available for download on Thursday, June 17, 2027

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