Recent scientific advances in the discovery of human pluripotent stem cells (hPSCs) and the ability to direct hPSCs to specialized cell types offer the potential to revolutionize the study of human development and disease, drug screening and safety profiling, and cell-based regenerative therapies. To realize this promise of hPSCs, however, efficient biomanufacturing platforms are needed to robustly generate large quantities of high quality hPSCs and their derivatives. Traditional biomanufacturing strategies employ bioreactors to control the environment of the cell, providing extrinsic signals that direct cell fates. Here, the investigators propose a novel intrinsic paradigm for control of hPSC fate, using recent advances in molecular biology to program an hPSC to develop into a heart muscle cell in the absence of external cues. Successful completion of this project will provide a deeper fundamental understanding of how hPSCs make decisions to turn into more specialized cells and provide a new strategy to biomanufacture specialized cells from stem cells by engineering the cell.
This project will develop a novel paradigm for controlling human pluripotent stem cell (hPSC) differentiation, using cell intrinsic sensing of cell state and control of developmental signaling pathways. The investigators will employ recent advances in understanding how canonical Wnt signaling drives hPSC commitment to mesoderm and subsequent specification to cardiac mesoderm and functional cardiomyocytes to design cells that sense their differentiation status and drive signaling to elicit desired differentiation trajectories. The investigators will identify mesoderm and cardiac mesoderm-specific promoter elements and use these to activate expression of Wnt pathway modulators, generating hPSC lines that differentiate to functional cardiomyocytes in the absence of extrinsic signaling. Then, the investigators will test the hypothesis that cell intrinsic control of differentiation enables more robust and scalable biomanufacturing of cardiomyocytes than extrinsic control via application of small molecules during scale down for high throughput screening application and scale up for production of cells suitable for clinical evaluation. The broader impacts of this project include providing a blueprint for engineering hPSC lines for biomanufacturing of somatic lineages beyond cardiomyocytes. Education and outreach activities will also stem cell engineers at the graduate and undergraduate levels, and will provide outreach to K-12 students, K-12 teachers, undergraduate students, and the general public on aspects of stem cell science and biomanufacturing.
