Activation of Diverse Signaling Pathways by Ex-Vivo Delivery of Multiple Cytokines for Myocardial Repair

作者：Konoplyannikov Mikhail; Haider Khawaja Husnain; Lai Vien Khach; Ahmed Rafeeq P H; Jiang Shujia; Ashraf Muhammad^*

来源：Stem Cells and Development, 2013, 22(2): 204-215.

DOI：10.1089/scd.2011.0575

We tested the hypothesis that simultaneous transgenic overexpression of a select quartet of growth factors activates diverse signaling pathways for mobilization and participation of various stem/progenitor cells for cardiogenesis in the infarcted heart. Human insulin growth factor-1 (IGF-1), vascular endothelial growth factor (VEGF), stromal cell-derived factor-1 (SDF-1a), and hepatocyte growth factor (HGF) plasmids were synthesized and transfected into skeletal myoblasts (SM) from young male wild-type or transgenic rats expressing green fluorescent protein (GFP). Overexpression of growth factors in transfected SM ((SM)-S-Trans) was confirmed by reverse transcription polymerase chain reaction, western blotting, and fluorescence immunostaining. Using our custom-made growth factor array and western blotting, multiple angiogenic and prosurvival factors were detected in (SM)-S-Trans, including secreted frizzled related protein-1,2,4,5, matrix metalloproteinases-3 and 9, connexin-43, netrin-1, Nos-2, Wnt-3, Akt, MAPK42/44, Stat3, nuclear factor kappa B (NFk kappa hypoxia-inducible factor 1 (HIF-1 alpha), and protein kinase C (PKC). The conditioned medium (CM) from (SM)-S-Trans was cytoprotective for cardiomyocytes following H2O2 treatment [P %26lt; 0.01 vs. CM from native SM ((SM)-S-Nat)], promoted a higher transwell migration of human umbilical cord vein endothelial cells (223.3 +/- 1.8, P %26lt; 0.01) and in vitro tube formation (47.8 +/- 1.9, P %26lt; 0.01). Intramyocardial transplantation of 1.5 x 10(6) (SM)-S-Trans (group-3) in a rat model of acute myocardial infarction induced extensive mobilization of cMet(+), ckit(+), ckit(+)/GATA(4+), CXCR4+, CD44(+), CD31(+), and CD59(+) cells into the infarcted heart on day 7 and improved integration of (SM)-S-Trans in the heart compared to (SM)-S-Nat (group 2) (P %26lt; 0.05). Extensive neomyogenesis and angiogenesis in group-3 (P %26lt; 0.01 vs. group-2), with resultant attenuation of infarct size (P %26lt; 0.01 vs. group-2) and improvement in global heart P %26lt; 0.01 vs. group-2) was observed at 8 weeks. In conclusion, simultaneous activation of diverse signaling pathways by overexpression of multiple growth factors caused massive mobilization and homing of stem/progenitor cells from peripheral circulation, the bone marrow, and the heart for accelerated repair of the infarcted myocardium.

• 出版日期2013-1