Cardiomyocytes from CCND2-overexpressing human induced-pluripotent stem cells repopulate the myocardial scar in mice: A 6-month study

Background: Cardiomyocytes that have been differentiated from CCND2-overexpressing human induced-pluripotent stem cells (hiPSC-CCND2(OE) CMs) can proliferate when transplanted into mouse hearts after myocardial infarction (MI). However, it is unknown whether remuscularization can replace the thin LV scar and if the large muscle graft can electrophysiologically synchronize to the recipient myocardium. Our objectives are to evaluate the structural and functional potential of hiPSC-CCND2(OE) CMs in replacing the LV thin scar. Methods: NOD/SCID mice were treated with hiPSC-CCND2(OE) CMs (i.e., the CCND2(OE) group), hiPSC-CCND2(WT) CMs (the CCND2(WT) group), or an equal volume of PBS immediately after experimentally-induced myocardial infarction. The treatments were administered to one site in the infarcted zone (IZ), two sites in the border zone (BZ), and a fourth group of animals underwent Sham surgery. Results: Six months later, engrafted cells occupied >50% of the scarred region in CCND2(OE) animals, and exceeded the number of engrafted cells in CCND2(WT) animals by similar to 8-fold. Engrafted cells were also more common in the IZ than in the BZ for both cell-treatment groups. Measurements of cardiac function, infarct size, wall thickness, and cardiomyocyte hypertrophy were significantly improved in CCND2(OE) animals compared to animals from the CCND2(WT) or PBS-treatment groups. Measurements in the CCND2(WT) and PBS groups were similar, and markers for cell cycle activation and proliferation were significantly higher in hiPSC-CCND2(OE) CMs than in hiPSC-CCND2(WT) CMs. Optical mapping of action potential propagation indicated that the engrafted hiPSC-CCND2(OE) CMs were electrically coupled to each other and to the cells of the native myocardium. No evidence of tumor formation was observed in any animals. Conclusions: Six months after the transplantation, CCND2-overexpressing hiPSC-CMs proliferated and replaced >50% of the myocardial scar tissue. The large graft hiPSC-CCND2(OE) CMs also electrically integrated with the host myocardium, which was accompanied by a significant improvement in LV function.