Dose-Response Effect of Cardiac-Inducing RNA(CIR) on the Differentiation of Human Fibroblast Cells into Cardiomyocytes.
Abstract: Acute myocardial infarction (AMI), commonly termed heart attack, is the leading cause of death worldwide and every year about 790,000 Americans are inflicted by this disease. In AMI, the coronary artery gets blocked causing cardiac muscle cell necrosis, due to deprivation of oxygen and nutrients, however the cells are subsequently replaced by non-contractile fibrous connective tissue. Since adult cardiac muscle cells lack regenerative ability, one potential approach to reconstitute normal heart function after AMI would be stem cell therapy. Earlier studies in our laboratory used cardiac-inducing RNAs (CIR) from normal embryos to restore heart function in cardiac non-function mutant salamanders. Similarly, by cloning and sequencing RNA libraries from human fetal heart (396 clones), two Human derived clones of Cardiac Inducing RNA (CIR) were obtained that have the ability to promote differentiation of non-cardiac cells into cardiomyocytes. Transfection of the CIR using liposomes further promotes differentiation of human dermal fibroblasts into cardiomyocytes in vitro. We hypothesize that CIR directs differentiation of human fibroblasts into cardiac lineage cells. Currently, we are performing dose-response experiments to determine optimal concentrations of CIR that promote efficient differentiation. To test our hypothesis, we are using polyethylene glycol liposome-mediated CIR transfection at varying concentrations. Human induced pluripotent stem cell (HiPSC) and Human cardiac and dermal fibroblast cells are co-transfected with a Luciferase reporter gene to measure transfection efficiency by Flow Cytometry. The differentiation into cardiomyocytes is measured using RT-PCR and immunofluorescent confocal microscopy to analyze the contractile protein and myofibril formation. One of the major treatment benefits would be, cells (fibroblasts) can be obtained from the patients themselves, which would be immunocompatible. Thus, this developing technology to replace damaged heart muscle cells with viable cardiomyocytes to restore heart function promises to provide a cure for heart failure with tremendous medical, humanitarian, and economic benefits to our society.
Judging Forms – Official judges only