Understanding Cardiac Progenitors to deliver Regenerative Medicine and Disease Modelling

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Heart disease, including damage to heart muscle, kills more people worldwide than any other illness. This means strategies for cardiac regeneration, such as cell replacement therapy after myocardial infarction and 'disease in a dish' modelling in vitro, are urgently needed. Access to cardiovascular cells including cardiomyocytes from human pluripotent stem cells (hPSCs) offers therapeutic potential capable of revolutionising cardiology but, as yet, the promise is unmet. Progress is currently hindered by our poor understanding of, and thus ability to control, cardiac cell development - principally the events of cardiac progenitor cell (CPC) specification, self-renewal and differentiation. The lack of comparative molecular benchmarking with human embryogenesis, required to authenticate cells in vitro, is another major barrier. By consequence, at present, mixtures of poorly specified cardiomyocyte identities emerge from uncontrolled, heterogeneous differentiation. The resultant cells are inadequate for accurate disease modelling (disease phenotypes are highly variable) and would be potentially life-threatening (arrhythmogenic) if transplanted. There are three major components to my study, delivery of which will unlock potential for academic research, clinical and industrial applications: 1) discovering the identity of CPCs and how to control them, 2) authenticating ventricular cardiomyocyte differentiation and 3) testing the system in an in vivo context.The challenge to unravelling the complexities of cardiac cell development is that differentiation is heterogeneous, CPCs are diverse, and cells progress along the differentiation trajectory at different rates. To overcome this, I have created an in vitro model system of single cell genomics linked to clonal lineage tracing and fate assessment. This integrated and unbiased approach will enable me to identify the gene expression and chromatin signatures pertinent to CPC self-renewal and cardiomyocyte differentiation potential. Perturbation experiments in vitro, combined with comparative measurements of the native cells in human embryogenesis, will help me identify the most functionally influential and authentic candidates. Together, these experiments will deliver the knowledge and tools to isolate, expand and differentiate the most desirable CPC identities - those with potential to be accurately programmed to specific target cell types e.g. ventricular cardiomyocytes (a key target for disease modelling and cell therapy). To learn how to accurately control downstream differentiation of CPCs to ventricular cardiomyocytes, I will use the gene expression and chromatin structure of human ventricular myocardium through development to benchmark differentiation in vitro. The relationship between signalling pathways and the regulation and expression of key transcription factors will be systematically resolved, to achieve the accurate, lineage-specific differentiation of CPCs. Finally, to validate these advances in CPC maintenance, tracking and differentiation to translational relevance, I will work in partnership with international collaborators to assess cell behaviour following transplantation into an animal model. In summary, by delivering research innovation of immediate clinical and industrial importance, this project is closely in tune with the UK Government's Industrial Strategy. The predicted advances will benefit applications in regenerative medicine, cardiac disease modelling, biomarker identification and drug screening. Moreover, it will also allow me to establish myself as a global leader in the field and, by partnering with industry, a persistent force towards achieving these translational goals.