Development of growth factor fusion proteins for cell-triggered drug delivery
Development of growth factor fusion proteins for cell-triggered drug delivery
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DOI：
10.1096/fj.00-0564fje
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发表时间：
2001-05-01
期刊：
影响因子：
4.8
通讯作者：
Hubbell, JA
中科院分区：
文献类型：
--
作者：
Sakiyama-Elbert, SE;Panitch, A;Hubbell, JA
本研究的目标是开发一种生长因子递送方法，该方法允许以与组织再生相关的局部酶活性可以触发生长因子释放的方式将生长因子稳定掺入细胞生长基质中。我们通过设计修饰的β-神经生长因子（β-NGF）融合蛋白并测试其促进神经突延伸的能力，在周围神经再生的背景下研究了这种方法。选择纤维蛋白作为细胞生长基质，并利用因子XIIIa的转氨酶活性将β-NGF融合蛋白共价掺入纤维蛋白基质中。重组表达了新型β-NGF融合蛋白，其含有外源性因子XIIIa底物以允许掺入纤维蛋白基质。在因子XIIIa底物和NGF结构域之间放置一个间插纤溶酶底物结构域，以允许细胞介导的生长因子响应于侵入细胞产生的纤溶酶而释放。与可溶性天然β-NGF相比，固定化的NGF融合蛋白与插入的功能性纤溶酶切割序列使胚胎鸡背根神经节的神经突延伸增加了50%，相对于没有NGF增加了350%。这些结果表明，这种新的生长因子递送方法，其中该因子根据细胞需求递送，可以增强神经再生，并可能在组织工程中有用。
The goal of this research was to develop an approach to growth factor delivery that would allow the stable incorporation of growth factors within a cell in‐growth matrix in a manner such that local enzymatic activity associated with tissue regeneration could trigger growth factor release. We investigated this approach in the context of peripheral nerve regeneration by designing modified beta‐nerve growth factor (β‐NGF) fusion proteins and testing their ability to promote neurite extension. Fibrin was selected as the cell in‐growth matrix, and the transglutaminase activity of factor XIIIa was utilized to covalently incorporate β‐NGF fusion proteins within fibrin matrices. Novel β‐NGF fusion proteins, which contained an exogenous factor XIIIa substrate to allow incorporation into fibrin matrices, were expressed recombinantly. An intervening plasmin substrate domain was placed between the factor XIIIa substrate and the NGF domain to allow cell‐mediated growth factor release in response to plasmin, which is generated by invading cells. Immobilized NGF fusion protein with an intervening functional plasmin cleavage sequence enhanced neurite extension from embryonic chick dorsal root ganglia by 50% relative to soluble native β‐NGF and by 350% relative to the absence of NGF. These results suggest that this novel approach to growth factor delivery, in which the factor is delivered upon cellular demand, could enhance nerve regeneration and may be useful in tissue engineering.