Nanowired human isogenic cardiac organoids to treat acute myocardial ischemia/reperfusion injuries - Project Summary: In the U.S., there are more than 735,000 myocardial infarctions (MI) each year. While percutaneous coronary intervention (PCI) has significantly reduced acute adverse repones, the long-term prognosis for post-ischemia/reperfusion (I/R) patients remains poor. Due to the limited regenerative capacity of human hearts, human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) have received significant attention due to their proven capacity to restore contractile function upon transplantation to injured hearts in various mammalian models, leading to multiple ongoing clinical trials. However, the current transplantation approach mainly relies on dissociated hPSC-CMs, leading to low cell survival, moderate functional improvement, arrhythmogenic risk, and poor scalability. To address these challenges, our lab developed nanowired, pre- vascularized human cardiac organoids composed of hPSC-CMs, human primary cardiac fibroblasts, endothelial cells, stromal cells, and electrically conductive silicon nanowires (e-SiNWs). Endothelial cells are used to induce vasculature formation within the organoids, and e-SiNWs are added to create an electrically conductive microenvironment to facilitate hPSC-CM contractile development and their electrical integration with the host myocardium. Our preliminary in vivo data showed that nanowired organoids illustrated robust hPSC- CM engraftment and superior functional recovery. The major barriers in their clinical translation include: 1) the use of animal proteins in the cell and organoid culture and 2) the lack of functional benefit demonstration in a large animal model. Replacing human primary cells with isogenic hPSC-derived cells for organoid fabrication would reduce batch-to-batch variations and enhance immune compatibility through Major Histocompatibility Complex (MHC) matching hPSC donors with human recipients. In addition, while the current hPSC-CM implantation strategy has been focused on intramyocardial injection, developing an effective approach for intracoronary delivery of the organoids will accelerate their clinical translation. The goal of this proposal is to develop clinical-grade hPSC cardiac organoids and demonstrate their functional benefits with a large animal model to generate enabling data for IND submission. The central hypothesis of this proposal is the nanowired isogenic hPSC cardiac organoids provide a scalable system to both efficiently and effectively implant hPSC-CMs for cardiac repair. The proposal is innovative in that we will 1) derive isogenic hPSC-derived cells in xeno-free, chemically defined conditions to develop clinical-grade cardiac organoids for implantation and 2) leverage the size and the endothelial lumen-like structures in the organoids to develop an effective intracoronary delivery strategy. Accordingly, we will pursue the following 2 aims: 1) Fabricate and characterize nanowired human cardiac organoids using isogenic cardiac cells derived from hPSCs in xeno-free, chemically defined conditions, and 2) Determine the therapeutic efficacy of the nanowired isogenic hPSC cardiac organoids with a porcine I/R (ischemia/reperfusion) model.
