Generating a 3D Micro necroptosis induced Myocardial Infarction model

Cardiovascular disease remains one of the leading causes of death worldwide, with more than 4 out of 5 of these due to heart attacks.

Following a myocardial infarction (MI) the loss of cardiomyocytes (CMs) is irreparable due to their non-proliferative state. Following a MI one of the most promising forms of recovery is using therapeutic drugs.

Since generating a new drug requires considerable amounts of money and animal testing there is a need for alternative models such as human induced pluripotent stem cell (hPSC) models. hPSCs provide an excellent platform to model cardiac disease, although 2D models are too immature and currently 3D models are not viable for screening of therapeutic compound libraries.

To solve this, I aim to implement a 'kill switch' in a subpopulation of a 3D cardiac tissue, that when triggered will induce necroptosis in the population, simulating a localized heart attack.

Integrating macrophages will improve the physiological relevance and cTnT-FUCCI CMs will be used to monitor CM proliferation.To generate this innovative, dependable, inducible model of MI using hiPSCs, the project is divided into 3 achievable goals.1.Generate a hPSC cell line with inducible kill switch system2.Integrate macrophages into microtissues, induce the kill switch, investigate response to localized cell death and cellular changes producing a scRNA-Seq timeline3.Incorporate cTnT-FUCCI cardiomyocytes to investigate cardiac regenerative therapiesI will receive state-of-the-art genetic modification training to generate the cell line (Goal 1), immune cell generation and integration (Goal 2) and bioinformatic study to identify key pathways identified through scRNA-Seq data generated from the MiniMI model to trial cardiac regenerative compounds (Goal 3).

This project will not only generate a reliable model of MI capable for screening of cardioprotective compounds, but also a malleable model suitable for easy adaptation to other researchers interests.