Abstract 13247: Preconditioning Cardiac Progenitor Cells With Biologically Active Gases (NO and CO) Enhances Cell Survival Through Activation of Survival Signal Pathways
Background: The poor survival of donor cell is one of the major limitations for effective stem cell therapy. Preconditioning cardiac progenitor cells (CPCs) with antioxidant or anti-apoptotic molecules may improve the stem cell survival and increase therapeutic efficacy. Nitric oxide (NO) and carbon monoxide (CO) are biologically active gases with increasingly well defined physiological roles in a number of disease states. Although breathing high concentrations of NO and CO is toxic, these gases protect against ischemia-reperfusion injury in the heart via anti-apoptotic effects. Therefore, we sought to determine whether preconditioning CPCs with these gases promotes cell survival following oxidative stress.
Methods and Results: We have recently established a protocol to purify c-kit positive, lin negative CPCs from the heart of patients. The results of lactate dehydrogenase (LDH) release assays demonstrated that CPCs preconditioned with the NO donor DETA-NOate and the heme oxygenase-1 (HO-1) inducer cobalt protoporphyrin (CoPP) promotes cell survival following oxidative stress induced by 1 mM H2O2 in basal medium, concomitant with significant up-regulation of HO-1 expression following preconditioning demonstrated by Western blot. CORM-3, a CO donor, also produced a cytoprotective effect compared with inactive CORM-3 (iCORM-3). Both the annexin V/PI assay and the caspase-3 activity assay showed that pretreatment of CPCs with low concentrations of NO and CO donors enhances the cell's resistance to apoptosis induced by oxidative stress. Fluorescence immunostaining showed nuclear translocation of nuclear factor-kappaB (NF-kappaB). Activation of phosphorylation of NF-kappaB, p38, Erk1/2, and STAT1/3 was also observed following preconditioning of CPCs with NO and CO donors.
Conclusions: Preconditioning of human CPCs with physiological concentrations of NO or CO promotes cell survival under oxidative stress. The mechanism appears to involve activation and nuclear translocation of NF-kappaB and activation of survival signal pathways such as the MAPK and ERK pathways. Our results may have important therapeutic implications for cardiac progenitor cell therapy in patients with myocardial infarction and ischemic heart disease.
- © 2010 by American Heart Association, Inc.