RECEPTOR KINASE GENE THERAPY FOR NEOINTIMAL HYPERPLASIA

受体激酶基因治疗新生内膜增生

基本信息

批准号：
6184883
负责人：
NEIL J. FREEDMAN
金额：
$ 33.94万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1999
资助国家：
美国
起止时间：
1999-07-01 至 2003-05-31
项目状态：
已结题

项目摘要

Percutaneous transluminal coronary angioplasty (PTCA) and coronary artery bypass graft surgery (CABG) are each used to treat approximately 400,000 coronary artery disease patients annually in the United States. PTCA and saphenous vein grafts in CABG are both complicated by proliferation of smooth muscle cells in the vessel subintima, in a process termed neointimal hyperplasia. Neointimal hyperplasia leads to significant coronary restenosis in approximately 30% of PTCA cases, and engenders significant vein graft stenoses--requiring some sort of intervention--in 30% of CABG cases within two years of surgery. The annual cost of PTCA restenosis alone exceeds $2 billion. This work proposes to test the hypothesis that transient gene therapy with G protein-coupled receptor kinases (GRKs) at the time of PTCA or saphenous in graft placement may substantially alleviate neointimal hyperplasia. GRKs are a family of enzymes which initiate either desensitization or signal termination for many G protein-coupled receptors, several of which have been implicated in the pathology of vascular smooth muscle cell proliferation and restenosis: endothelin A and B, angiotensin II (type I), thrombin, and thromboxane A2 receptors. Over-expression of GRK2 can drastically reduce signaling through all of these receptors in transfected cell systems. Systematically administered antagonists of the endothelin and angiotensin II (type I) receptors significantly reduce neointimal hyperplasia in animal models of vascular injury. This proposal therefore seeks to develop recombinant adenoviruses for expressing GRKs 2, 3, and 5 transiently in smooth muscle cells, and to test each of these recombinant viruses in rabbit and human smooth muscle cells for the ability to reduce cellular responses to agonists for the receptors listed above, as assessed by inositol phosphate signaling, cellular migration, DNA synthesis, and cell proliferation. Subsequently, GRK adenoviruses and adeno-associated viruses will be used ex vivo to treat rabbit jugular veins, which will be grated across the rabbit's ligated carotid artery in a well-established model of neointimal hyperplasia. Four weeks after surgery, vein grafts will be evaluated to determine the GRK's ability (a) to attenuate neointimal hyperplasia in vivo, by histology, and (b) to attenuate agonist-stimulated hyperplasia by transiently over-expressed GRKs in the rabbit model of vascular injury would have therapeutic implications for restenosis in PTCA and vein graft failure in CABG.

在美国，经皮易流性冠状动脉血管成形术（PTCA）和冠状动脉搭桥移植手术（CABG）每年用于治疗约40万名冠状动脉疾病患者。 CABG中的PTCA和隐静脉移植物在容器亚米塔中的平滑肌细胞的增殖变得复杂，这在称为新内膜增生的过程中。新内膜增生会导致大约30％的PTCA病例导致冠状动脉再狭窄，并在手术后的两年内导致30％的CABG病例，引起了明显的静脉移植stenose（要求进行某种干预）。仅PTCA再狭窄的年度成本超过20亿美元。这项工作提出了以下假设：在PTCA或在移植物放置中使用G蛋白偶联受体激酶（GRK）的瞬时基因治疗可能会大大减轻新的新膜增生。 GRK是一个酶家族，用于许多G蛋白偶联受体启动脱敏或信号终止，其中几种与血管平滑肌细胞增殖和再吻合性的病理有关：内皮素A和B，血管素II（I型I），凝血酶II），螺栓蛋白和Thromboxane A2受体。 GRK2的过表达可以通过转染的细胞系统中的所有这些受体大大减少信号传导。在血管损伤的动物模型中，系统施用内皮素和血管紧张素II（I型）受体的拮抗剂可显着降低新内膜增生。 This proposal therefore seeks to develop recombinant adenoviruses for expressing GRKs 2, 3, and 5 transiently in smooth muscle cells, and to test each of these recombinant viruses in rabbit and human smooth muscle cells for the ability to reduce cellular responses to agonists for the receptors listed above, as assessed by inositol phosphate signaling, cellular migration, DNA synthesis, and cell proliferation.随后，将使用GRK腺病毒和与腺相关的病毒用于治疗兔子颈静脉，该静脉将在兔子绑扎的颈动脉上磨碎，以良好的新内膜增生模型。 Four weeks after surgery, vein grafts will be evaluated to determine the GRK's ability (a) to attenuate neointimal hyperplasia in vivo, by histology, and (b) to attenuate agonist-stimulated hyperplasia by transiently over-expressed GRKs in the rabbit model of vascular injury would have therapeutic implications for restenosis in PTCA and vein graft failure in CABG.