The angiogenic gene profile of circulating endothelial progenitor cells from ischemic stroke patients

Abstract

Altres ajuts: AR is supported by the Miguel Servet programme (CP09/00265) from the Spanish Ministry of Health (Instituto de Salud Carlos III). This work has been funded by Instituto de Salud Carlos III, grant PI10/00694 and the Spanish stroke research network RENEVAS (RD06/0026/0010) co-financed by the European Regional Development Fund (ERDF). This work has been supported also by the European Union's Seventh Framework Programme (FP7/2007-2013) under grant agreements n° 201024 and n° 202213 (European Stroke Network).The identification of circulating endothelial progenitor cells (EPCs) has introduced new possibilities for cell-based treatments for stroke. We tested the angiogenic gene expression of outgrowth endothelial cells (OECs), an EPC subtype capable to shape vessel structures. OECs (at colony or mature stages) from ischemic stroke patients (n=8) were characterized using the RT 2 Profiler TM human angiogenesis PCR Array, and human microvascular endothelial cells (hCMEC/D3) were used as an expression reference of endothelial cells. Colony-OECs showed higher expression of CCL2, ID3, IGF-1, MMP9, TGFBR1, TNFAIP2, TNF and TGFB1. However, BAI-1, NRP2, THBS1, MMP2 and VEGFC expression was increased in mature-OECs (p<0.05). ID3 (p=0.008) and TGFBR1 (p=0.03) genes remained significantly overexpressed in colony-OECs compared to mature-OECs or hCMEC/D3. MMP9 levels were significantly increased in colony-OECs (p=0.025) compared to mature-OECs. Moreover, MMP-2, VEGF-C, THBS1 and NRP-2 gene expression was also significantly increased in mature-OECs compared to hCMEC/D3 (p<0.05). Some of these genes were positively validated by RT-PCR. Our study shows that OECs from stroke patients present higher levels of pro-angiogenic factors at early stages, decreasing in mature OECs when they become more similar to mature microvascular endothelial cells