Research
Title: Modelling endocardial and coronary endothelial cells using pluripotent stem cells
Abstract
Endocardial endothelial cells lining the heart chambers contribute to key cardiac functions. These include revascularization of the heart after myocardial infarction (MI), the formation of heart valves and paracrine signalling affecting myocardial function. Currently there are no endocardial cell lines available. Therefore, there is a clinical need for easily accessible and defined endocardial-like population of cells for modelling endocardial cell biology, cell dysfunction and for translational applications such as in regenerative medicine.
I have developed a serum-free, chemically-defined, high yield protocol to differentiate pluripotent stem cells (PSCs), into cells enriched for endothelial and endocardial specific markers. The cells generated with this protocol also express these markers at significantly higher levels than cells differentiated using other PSC endothelial differentiation protocols and endothelial cells (ECs) isolated from adult and foetal tissues. The gene expression profile of the cells generated is important as this will impact the function of the cells generated. The endocardial-like cells generated were transitioned with VEGF-A, in a dose dependent manner, into cells with a transcriptional phenotype resembling a coronary EC identity. The endocardiaI-like cells have also been shown to improve engineered heart tissue function. In future work, the endocardial-to-coronary EC transition model will be exploited to identify pathways and compounds that can enhance the transition of endocardial cells into coronary ECs and thus might be used to improve re-vascularisation and cardiac function in vivo post-MI and in other cardiac diseases.
After the generation and functional characterisation of iPSC-endocardial cells, I next explored how this work might be translated towards cellular transplantation post-MI to improve cardiac function. I worked on generating iPSCs using a novel RNA-based reprogramming protocol combined with late-outgrowth endothelial progenitor cells. This work could now lay the foundations for reprogramming of iPSCs under conditions suited for clinical translation and thereby the potential use of iPSC-endocardial cells in the clinic for the future treatment of cardiovascular diseases.
Aims
- To define a pluripotent stem cell (PSC) differentiation protocol that generates endothelial-like cells that have an enriched expression for endocardial genes (endocardial-like)
- Functional characterisation of iPSC-derived endocardial-like cells
- Combining self-replicative RNA reprogramming with late-outgrowth endothelial progenitor cells (EPCs) to produce iPSCs