Activation of diverse signaling pathways by Ex-vivo delivery of multiple cytokines for myocardial repair

Mikhail Konoplyannikov, Khawaja Husnain Haider, Vien Khach Lai, Rafeeq P.H. Ahmed, Shujia Jiang, Muhammad Ashraf

Research output: Contribution to journalArticle

20 Scopus citations

Abstract

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 (TransSM) 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 TransSM, 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 (NFκB), hypoxia-inducible factor 1 (HIF-1α), and protein kinase C (PKC). The conditioned medium (CM) from TransSM was cytoprotective for cardiomyocytes following H 2O2 treatment [P<0.01 vs. CM from native SM ( NatSM)], promoted a higher transwell migration of human umbilical cord vein endothelial cells (223.3±1.8, P<0.01) and in vitro tube formation (47.8±1.9, P<0.01). Intramyocardial transplantation of 1.5×106 TransSM (group-3) in a rat model of acute myocardial infarction induced extensive mobilization of cMet+, ckit+, ckit+/GATA4+, CXCR4+, CD44+, CD31+, and CD59+ cells into the infarcted heart on day 7 and improved integration of TransSM in the heart compared to NatSM (group 2) (P<0.05). Extensive neomyogenesis and angiogenesis in group-3 (P<0.01 vs. group-2), with resultant attenuation of infarct size (P<0.01 vs. group-2) and improvement in global heart function (P<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.

Original languageEnglish (US)
Pages (from-to)204-215
Number of pages12
JournalStem Cells and Development
Volume22
Issue number2
DOIs
StatePublished - Jan 15 2013

ASJC Scopus subject areas

  • Hematology
  • Developmental Biology
  • Cell Biology

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