PREVENTION OF VEIN GRAFT SPASM

描述(由申请人提供)： 血管痉挛发生在用于血管搭桥手术的血管中。这种痉挛是由于平滑肌松弛受损造成的。松弛是由环核苷酸依赖的信号通路介导的。我们已经证明，一个小的热休克蛋白HSP20的磷酸化代表了一个点，在这个点上，环核苷酸信号通路会聚在一起，导致松弛。这项研究的假设是，包括HSP20基序和蛋白转导结构域的血管松弛分子将通过更好地保存移植物在收获期间和通过防止内膜增生来防止1)由于血管痉挛而导致的短期静脉移植物失败和2)长期移植物失败。这项研究的具体目的是：#1：构建连接到蛋白转导结构域(PTD)的重组HSP20，并确定该工程蛋白是否在体外逆转人类血管平滑肌痉挛。#2：确定PTD-HSP20对与内膜增生相关的动态细胞骨架过程的影响。#3：确定HSP20类似物在体内移植静脉的蛋白质转导的可行性。该项目的目标是设计生物分子，通过将内源性松弛蛋白(HSP20)的类似物直接引入到血管中来增强静脉移植物的保存。静脉移植物是这些治疗方法的理想靶点，因为移植物可以在体外进行治疗，从而为工程蛋白/肽疗法的本地化提供了最佳的环境。这代表了一种新的方法，因为受体和信号级联被“绕过”，效应蛋白/肽直接进入靶细胞。这代表了设计具有生物活性的蛋白质/多肽分子的“后基因组”平台。

DESCRIPTION (provided by applicant): Vasospasm occurs in the blood vessels used for vascular bypass operations. This spasm is due to impaired relaxation of the smooth muscle. Relaxation is mediated by cyclic nucleotide-dependent signaling pathways. We have demonstrated that the phosphorylation of a small heat shock protein, HSP20 represents a point in which the cyclic nucleotide signaling pathways converge to cause relaxation. The hypotheses of this investigation are that vasorelaxing molecules, engineered to include HSP20 motifs and protein transduction domains, will prevent 1) short term vein graft failure due to vasospasm and 2) long term graft failure by better preservation of the graft during harvest and by the prevention of intimal hyperplasia. The specific aims of this investigation are: #1: Generate recombinant HSP20 linked to a protein transduction domain (PTD) and determine if this engineered protein reverses human vascular smooth muscle spasm ex vivo. #2: Determine the effect of PTD-HSP20 on dynamic cytoskeletal processes relevant to intimal hyperplasia. #3: Determine the feasibility of protein transduction of HSP20 analogues of vein grafts in vivo. The goal of this project is to engineer biomolecules that enhance vein graft preservation by directly introducing the analogues of an endogenous relaxing protein (HSP20) into the smooth muscle. Vein grafts represent an ideal target for these therapeutic approaches in that the graft can be treated ex vivo, thus providing an optimal environment for the localization of engineered protein/peptide therapeutics. This represents a novel approach in that the receptors and signaling cascades are "bypassed" and the effector protein/peptide is directly introduced into the target cell. This represents a "post-genomic" platform for engineering biologically active protein/peptide molecules.