Abstract 479: Cardiomyopathy Phenotypes Observed in Human Engineered Heart Tissue Depend on Functional Maturation

Abstract

Human iPSC-based models of cardiomyopathies have become increasingly prevalent. These models enable the study of genotype-phenotype relationships within a human context and have already led to new insights. A potential limitation of iPSC-derived cardiomyocytes (iPSC-CMs) is their varying degrees of functional maturation with respect to electrophysiology, calcium handling, and contractility. Variable levels of maturity may explain how different groups investigating hypertrophic cardiomyopathy (HCM) mutations in iPSC cardiomyocytes have arrived at conflicting results. Nevertheless, whether or not advanced functional maturation truly impacts disease phenotype in iPSC cardiomyocyte models remains an open question. Using a novel protocol that combines pacing and physiological media in a three-dimensional engineered heart tissue (EHT) derived from decellularized myocardium, we are able to mature human engineered heart tissues within three weeks to recapitulate key functional characteristics of adult heart tissue, such as a robustly positive force-frequency response, a strong post-rest potentiation, potent calcium handling and excitation-contraction handling, and mature isometric twitches that closely match that of intact adult human ventricular myocardium. Using our platform, we investigated the effect of maturation on the functional consequence of a classic variant (E62Q) in the thin filament protein alpha-tropomyosin (TPM1). Initial characterization of TPM1 E62Q in EHTs show a hypercontractile phenotype compared to its isogenic wild-type line, with limited changes in calcium handling. Preliminary data on maturing TPM1 E62Q EHTs show that additional hallmarks of disease may develop with maturation such as arrhythmogenicity, hypertrophy, and fibrosis. In our work, we develop a method for advanced maturation of hiPSC-CMs in EHTs and show the dependence on maturation status of progressive hallmarks of HCM in human EHTs compared to EHTs grown under conditions standard for the field.