Establishment of a functionally active collagen-binding vascular endothelial growth factor fusion protein in situ
Establishment of a functionally active collagen-binding vascular endothelial growth factor fusion protein in situ
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DOI：
10.1161/01.atv.0000233359.74484.77
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发表时间：
2006-09-01
影响因子：
8.7
通讯作者：
Asahara, Takayuki
中科院分区：
文献类型：
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作者：
Ishikawa, Tetsuya;Eguchi, Masamichi;Asahara, Takayuki
目的 - 组织再生需要生长因子和胶原蛋白等细胞外基质，作为细胞生长的支架。我们建立了由纤连蛋白胶原结合域 (FNCBD) 和血管内皮生长因子 (VEGF) 121 组成的 FNCBD-VEGF121，并研究了其特性。方法和结果 - FNCBD-VEGF121 特异性结合明胶和 I、II、III、IV 和 V 型胶原。这种胶原蛋白结合的 FNCBD-VEGF121 捕获可溶性 VEGF 受体 2 (VEGFR-2)/Fc 嵌合蛋白。 FNCBD-VEGF121 的细胞生长促进活性与 VEGF121 几乎相同。 VEGF 融合蛋白显着增强脐带血来源的内皮祖细胞 (EPC) 上 VEGFR-2 的表达 (71.6 +/- 0.8%)。预期的是，胶原蛋白结合的 VEGF 融合蛋白不仅促进内皮细胞 ( EC) 的生长，而且还诱导从骨髓 CD34(+) 细胞扩增的非贴壁细胞上 VEGFR-2 ( 63.7 +/- 0.8%) 的表达。此外，VEGF 融合蛋白增强了基质胶模型中 EC 的芽形成。体内实验表明，FNCBD-VEGF121 对 EPC 动员具有局部效应，但没有全身效应。结论 - 这些结果表明，FNCBD-VEGF121 与胶原基质一起稳定地维持 VEGF 的最佳局部高浓度，并原位刺激 EC 和 EPC，提供血管再生生态位。
Objective - Tissue regeneration requires both growth factor and extracellular matrix such as collagen, serving as a scaffold for cell growth. We established FNCBD-VEGF121, consisting of the fibronectin collagen-binding domain ( FNCBD) and vascular endothelial growth factor ( VEGF) 121, and investigated its properties.Methods and Results - FNCBD- VEGF121 specifically bound to gelatin and type I, II, III, IV, and V collagen. This collagen-bound FNCBD- VEGF121 captured soluble VEGF receptor 2 ( VEGFR-2)/Fc chimeric protein. Cell growth-promoting activity of FNCBD- VEGF121 was almost identical to that of VEGF121. The VEGF fusion protein significantly enhanced the expression of VEGFR-2 ( 71.6 +/- 0.8%) on endothelial progenitor cells ( EPCs) derived from umbilical cord blood. Expectably, the collagen-bound VEGF fusion protein not only promoted the growth of endothelial cells ( ECs) but also induced the expression of VEGFR-2 ( 63.7 +/- 0.8%) on non-adherent cells expanded from bone marrow CD34(+) cells. Moreover, the VEGF fusion protein enhanced sprout formation of ECs in a matrigel model. In vivo experiments revealed that FNCBD- VEGF121 had local effects but not systemic effect on EPC mobilization.Conclusions - These results suggest that FNCBD- VEGF121 stably maintains an optimally high and local concentration of VEGF with collagen matrix and stimulates both ECs and EPCs in situ, supplying a vascular regeneration niche.