Background and Objective: Reprogramming of somatic fibroblasts into induced cardiac progenitor cells (iCPCs) has provided a promising therapeutic paradigm for cardiac repair. This study aims to generate new iCPCs using CRISPR technology and evaluate their therapeutic potential in a myocardial infarction (MI) animal model.
Methods: Tail-tip fibroblasts (TTFs) were isolated from Nkx2-5RFP tracing mice and induced using the CRISPR activation system targeting 10 progenitor genes. iCPCs were implanted into mouse infarcted myocardium. Heart function was measured by echocardiography and heart tissue sections were harvested for immunostaining.
Results: Generation of CRISPR-iCPC clones labeled with RFP (Fig. 1A) was confirmed by expression of early heart development markers. Ejection fraction and fractional shortening of MI mice were significantly improved after transplantation of iCPCs as compared to the control groups (Fig. 1B). Masson’s staining also showed that the infarct area was reduced in groups with implanted iCPCs (Fig. 1C). Finally, it was confirmed that engrafted iCPCs (RFP+) can differentiate into cTnT+ cardiomyocytes, α-SMA+ smooth muscle cells, or CD31+ endothelial cells in the infarcted heart (Fig. 1D).
Conclusion: CRISPR-iCPCs improved cardiac function of infarcted heart and can serve as a novel therapeutic for regenerative cell therapy in the future.