The Activation of the LIMK/Cofilin Signaling Pathway via Extracellular Matrix–Integrin Interactions Is Critical for the Generation of Mature and Vascularized Cardiac Organoids

The generation of mature and vascularized human pluripotent stem cell-derived cardiac organoids (hPSC-COs) is necessary to ensure the validity drug screening disease modeling. This study investigates effects cellular aggregate (CA) stemness self-organization on hPSC-COs elucidates mechanisms underlying organoid (CO) maturation vascularization. COs derived from 2-day-old CAs with high (H-COs) 5-day-old low (L-COs) were generated in a self-organized microenvironment via Wnt signaling induction. finds that H-COs exhibit ventricular, structural, metabolic, functional cardiomyocyte vessel networks consisting endothelial cells, smooth muscle pericytes, basement membranes compared L-COs. Transcriptional profiling shows upregulation genes associated formation Through experiments LIMK inhibitors, activation ROCK-LIMK-pCofilin ECM–integrin interactions leads H-COs. Furthermore, LIMK/Cofilin pathway induces TGFβ/NODAL PDGF for vascularization demonstrates first time axis plays an important role COs.