Novel zinc finger transcription factor ZFP580 promotes differentiation of bone marrow-derived endothelial progenitor cells into endothelial cells via eNOS/NO pathway.

ABSTRACT

BACKGROUND: The differentiation of endothelial progenitor cells (EPCs) plays a pivotal role in endothelial repair and re-endothelialization after vascular injury. However, the underlying mechanisms still remain largely elusive. Here, we investigated the role of the novel C2H2 zinc finger transcription factor ZFP580 in EPC differentiation and the molecular mechanisms behind EPC-mediated endothelial repair. METHODS: Bone marrow-derived EPCs were isolated, cultured, and identified. EPCs were infected with an adenovirus encoding ZFP580 or Ad-siRNA to silence ZFP580. Fluorescence-activated cell sorting (FACS) analysis was performed to analyze EPC surface makers. The expression of ZFP580, eNOS, VEGFR-2, CD31, CD34, CD45 and vWF was performed by Q-PCR, Western blot and immunostaining. NO donor SNAP or NOS inhibitor L-NAME was used to elucidate the possible molecular mechanism. Tube formation in vitro and angiogenesis assay in vivo were also used in this study. RESULTS: Both ZFP580 and eNOS were displayed dynamic expression during EPC differentiation. Overexpression of ZFP580 enhanced EPC differentiation, while knockdown suppressed it. ZFP580 also enhanced eNOS expression, and eNOS inhibition suppressed differentiation. Upregulation/knockdown of ZFP580 also enhanced/reduced endothelial tube formation from EPC in vitro, and angiogenesis in vivo in response to Matrigel plugs containing EPC. CONCLUSIONS: ZFP580 promotes not only the differentiation of EPCs into ECs by increasing the expression of eNOS and the availability of nitric oxide, but also the vessel formation in vitro and in vivo. This might represent a novel mechanism of ZFP580 in EPC differentiation and its therapeutic value in the treatment of vascular disease.