Suicide Gene Reveals the Myocardial Neovascularization Role of Mesenchymal Stem Cells Overexpressing CXCR4 (MSC^CXCR4)

<div><p>Background</p><p>Our previous studies indicated that MSC^CXCR4 improved cardiac function after myocardial infarction (MI). This study was aimed to investigate the specific role of MSC^CXCR4 in neovascularization of infarcted myocardium using a suicide gene approach.</p><p>Methods</p><p>MSCs were transduced with either lentivirus-null vector/GFP (MSC^Null as control) or vector encoding for overexpressing CXCR4/GFP. The MSC derived-endothelial cell (EC) differentiation was assessed by a tube formation assay, Dil-ac-LDL uptake, EC marker expression, and VE-cadherin promoter activity assay. Gene expression was analyzed by quantitative RT-PCR or Western blot. The suicide gene approach was under the control of VE-cadherin promoter. <i>In vivo</i> studies: Cell patches containing MSC^Null or MSC^CXCR4 were transduced with suicide gene and implanted into the myocardium of MI rat. Rats received either ganciclovir (GCV) or vehicle after cell implantation. After one month, the cardiac functional changes and neovascularization were assessed by echocardiography, histological analysis, and micro-CT imaging.</p><p>Results</p><p>The expression of VEGF-A and HIF-1α was significantly higher in MSC^CXCR4 as compared to MSC^Null under hypoxia. Additionally, MSC^CXCR4 enhanced new vessel formation and EC differentiation, as well as STAT3 phosphorylation under hypoxia. STAT3 participated in the transcription of VE-cadherin in MSC^CXCR4 under hypoxia, which was inhibited by WP1066 (a STAT3 inhibitor). In addition, GCV specifically induced death of ECs with suicide gene activation. <i>In vivo</i> studies: MSC^CXCR4 implantation promoted cardiac functional restoration, reduced infarct size, improved cardiac remodeling, and enhanced neovascularization in ischemic heart tissue. New vessels derived from MSC^CXCR4 were observed at the injured heart margins and communicated with native coronary arteries. However, the derived vessel networks were reduced by GCV, reversing improvement of cardiac function.</p><p>Conclusion</p><p>The transplanted MSC^CXCR4 enhanced neovascularization after MI by boosting release of angiogenic factors and increasing the potential of endothelial differentiation.</p>