Abstract 10664: Ephrin-B2 Treatment Restores the Efficiency of Peripheral Blood Mononuclear Cells Based-Therapy in Diabetes
We hypothesized that the acute treatment of peripheral blood mononuclear cells from diabetic patients (PB-MNC) with ephrin-B2 (EFNB2) enhances their proangiogenic potential in diabetic nude mice with hindlimb ischemia. Paw skin blood flow, angiographic score and capillary density were significantly increased by 95%, 110% and 97% in ischemic leg of diabetic mice receiving intravenous injection of EFNB2-stimulated PB-MNC compared to that of non-treated diabetic PB-MNC (n=12, p<0.001). Flow cytometry analysis evidenced that EFNB2 could bind to 5% of PB-MNC, including monocytes (70%), T lymphocytes (15%) and CD34+/CD45+ cells (15%). The EFNB2-related stimulation increased the ability of PB-MNC to bind to HUVEC monolayer through PI3 kinase-related pathway (n=6, p<0.001). Moreover, transcriptomic analysis demonstrated that EFNB2 stimulation rapidly increased the expression of proinflammatory/proangiogenic factors such as IL-1, MCP-1, and MIPs. Our findings that EFNB2 treatment increased PB-MNC-release of factors known to mobilize HSC and EPC from bone marrow prompted us to investigate the effects of intravenous administration of EFNB2-pretreated PBMNC on mobilization and differentiation of bone marrow-derived cells from host tissues. Administration of EFNB2-stimulated PB-MNC raised by 93% the number of circulating CD34+/VEGFR2 vascular progenitors in diabetic nude mice (n=9, p<0.001) and increased by 110% the ability of BM-derived cells of host mice to differentiate into cells with endothelial phenotype and promote post-ischemic revascularization (enhanced by 25%, 52% and 116% the paw skin blood flow, angiographic score and capillary density respectively, n=6, p<0.001). Therefore, EFNB2 treatment abrogates the diabetes-induced stem/progenitor cells dysfunctions and opens the way for the clinical development of an innovative and more accessible strategy based on the intravenous autologous transplantation of pretreated PB-MNC in patients with critical ischemic diseases.
- © 2011 by American Heart Association, Inc.