Human Heart Organoid-derived Extracellular Vesicles for Cardiac Intercellular Communication

Extracellular vesicles (EVs) are essential mediators of intercellular communication that can affect physiological and pathological conditions by transferring nucleic acids and proteins. In this study, we investigate EV-mediated intercellular communication using both in-silico and in-vitro models. We isolated EVs from human heart organoids (HHO); these systems reproduce many of the structural and functional aspects of the early developing human heart. These HHOs are grown under high glucose and high insulin conditions to model pregestational diabetes, a maternal disease associated with congenital heart defects. Using the diabetic HHO model, we hypothesized that EV miscommunication may be, in part, a cause of congenital heart defects associated with pregestational diabetes. EV characterization was done through Western blotting and nanoparticle tracking analysis (NTA). The anomalous diffusion and the biodistribution of EVs in a cardiac extracellular matrix (ECM) were modeled using 3-dimensional partial differential equations, and the model was verified by particle-based simulations. Our in-vitro research identified Alix and Flotinin-1 proteins as markers of EVs from diabetic HHO, and NTA revealed that EVs with a diameter of 105 nm are detected most frequently. Our in-silico research demonstrates that EV concentrations in the cardiac ECM exhibit varying patterns based on our NTA characterization. Our findings can provide a foundation for future EV-mediated treatment methods and improve our understanding of intercellular communication in pregestational diabetes and heart development.