TGF-ß Signalling Drives Chemotaxis of Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes in Response to MI Stimulus

Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) hold significant promise for cardiac regeneration therapies. However, the efficacy of such treatments depends on the ability of transplanted cells to migrate and integrate into the damaged myocardium, a process that remains poorly understood. In this study, we investigated the migratory behaviour of hiPSC-CMs using homogenised rat MI tissue to simulate myocardial infarction (MI) in vitro. Transwell migration assays demonstrated a concentration-dependent chemotactic response, with hiPSC-CM migration increasing up to threefold toward MI tissue homogenate. Wound healing assays further confirmed enhanced migration under MI-mimetic conditions. Bulk RNA sequencing revealed activation of the TGF-ß signalling pathway as a key regulator of this response. Inhibition of TGF-ß signalling, both pharmacologically and through antibody neutralisation, significantly reduced hiPSC-CM migration. These findings uncover a previously underappreciated chemotactic capability of hiPSC-CMs and identify TGF-ß signalling as a central mediator, offering new mechanistic insights and potential therapeutic targets to improve the integration and efficacy of hiPSC-CM-based cardiac regeneration strategies. Overall design: RNA-seq of day 15 hiPSC-CMs derived from the KUXP (WTSIi008-A) and QOLG (WTSli004-B) hiPSC lines, both migrated (KUXPM, QOLGM) and non-migrated (KUXPNONM, QOLGNONM) samples. HiPSC-CMs were cultured on transwell inserts with in the lower chamber medium supplemented with left ventricular (LV) homogenate derived from myocardial infarction (MI)–operated rat hearts. After 24 hours of incubation, migrated and non-migrated hiPSC-CMs were collected separately and processed for RNA sequencing.