A novel approach of generating and epigenomic characterization of cardiomycytes from human peripheral blood cells
Description: Due to the limited adult cardiomyocyte (CM) regeneration after cardiac injury, myocardial infarction (MI) patients usually suffer from permanent CM loss and eventually heart failure. Various approaches were developed recently aiming to enhancing cardiac tissue regeneration and regaining of normal function, nevertheless, currently, cardiac cell reprogramming and cell-based therapies are still far from a clinical practice, with much less satisfied outcomes in clinical trials. Particularly, one major concern of the current iPSC-based therapy is the possible formation of teratomas due to the viral integrations into the human genome and tumorigenesis of the residual undifferentiated iPSCs. A direct reprogramming of fibroblasts into CMs was attempted recently, but with a very low efficiency. Another problem is the lack of understanding of the molecular mechanisms underlying CM regeneration. Thus, there is an urgent need to develop a better method of cell reprogramming into CMs. We recently developed a novel cell reprogramming method by directly converting human peripheral blood mononuclear cells (PBMCs) into induced mesenchymal stem cells using episomal vectors, which are only presented shortly during the cell reprogramming process with low likelihood to be integrated into the human genome. Based on this, our two specific aims are: 1) Develop a novel non-integrating method of directly converting human PBMCs into CMs using a set of combined factors. 2) Characterize the epigenomic and single-cell transcriptomic landscapes of the direct conversion of human PBMCs into CMs. Our proposed studies are novel as: (1) it fills in a major gap in current knowledge with mechanistic approaches and methodologies; (2) Using the new cutting-edge NGS-based technologies such as RRBS, ATAC-seq and scRNA-seq in a novel CM programming method, we will be able to gain a deep understanding on the molecular mechanisms of CM reprogramming; (3) Our novel method will use the factors cloned in non-integrating vectors, thus eliminating the possibility of viral integration into the genome and the development of teratomas. The impact of our study is high. The findings will not only significantly advance our knowledge of the molecular mechanisms underlying the epigenomic regulation of gene expression at single-cell resolution in cardiac cell programming, but also improve our current cardiac cell therapy approach.
Funder: American Heart Association
Amount: $200,000
Funding type: research
Funder scheme: Innovative Project Award
Start date: 2018-07-01
End date: 2020-06-30
Investigators: Charles Wang, Charles Wang
Institution: Loma Linda University
Funded outputs count: 13
Funder award ID: 18IPA34170301
OpenAlex ID: G8946239403