Title:
Single Cell RNA-Sequencing of Cardiac Progenitor Cells Across Patient Age Populations
Single Cell RNA-Sequencing of Cardiac Progenitor Cells Across Patient Age Populations
Author(s)
Jayaraman, Arun Ravi
Advisor(s)
Davis, Michael E.
Gibson, Greg
Qiu, Peng
Cho, Hee Cheol
Gibson, Greg
Qiu, Peng
Cho, Hee Cheol
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Abstract
Human c-kit+ cardiac progenitor cells (CPCs) have seen success in the treatment of heart failure and myocardial dysfunction. However, research has demonstrated the reparative capacity of CPCs appears to be linked to the age of the patient, with younger patients having increased heart function and reduced fibrosis following treatment. We hypothesized that these differences may be driven by differing subtypes of CPCs existing in each donor sample. Using the high-resolution capabilities of single cell RNA-sequencing technologies, we hope to elucidate the different subtypes that may be giving rise to the differences in therapeutic outcomes observed during in vivo studies. In the first study we analyzed the differences between adult CPCs (aCPCs) and neonatal CPCs (nCPCs). In vivo studies indicated injected aCPC had reduced cell retention and cell proliferation due to increased phagocytosis in comparison to nCPCs. We found three distinct subtypes of CPCs following analysis of the data. The two nCPC-enriched clusters correlated strongly with wound healing and cell proliferation, while the third aCPC-enriched cluster indicated some immune response activity. Analysis of selected gene expression in the third cluster indicated reduced expression of CD47, an important anti-phagocytic protein, along with reduced expression of several important growth factor and ECM proteins.6 In the second study we analyzed the differences between pediatric patient populations. Previously published in vivo and in vitro results indicate reduced fibrosis and immune response and increased chemotaxis when using nCPCs in comparison to child CPCs (cCPCs). Subcluster analysis finds cCPC-enriched clusters upregulated in several fibrosis- and immune response-related genes. Clustering of genes indicates genes correlated in chemotaxis to be upregulated in nCPC clusters. We identified the surface proteins versican and ITGA2 to be upregulated in fibrosis-related cluster 6 cells. Flow cytometric analysis using antibodies specific to these proteins identified a cell population with high levels of both proteins, consistent with the gene expression profile identified by the cluster 6 cells. We hope that this research will allow for researchers in the future to better optimize for and predict clinical outcomes prior to injection in autologous CPC-based therapies.
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Date Issued
2021-12-08
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