TARGETING G PROTEINS IN VASCULAR INTIMAL HYPERPLASIA

靶向血管内膜增生中的 G 蛋白

• 批准号: 6165783
• 负责人: Walter J. Koch
• 金额: $ 34.65万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-08-01 至 2004-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6165783
• 关键词: Adenoviridae; G protein; G protein coupled receptor kinase; adeno associated virus group; biological signal transduction; blood vessel prosthesis; carotid artery; cell growth regulation; cell migration; cell proliferation; disease /disorder model; enzyme activity; enzyme inhibitors; heart revascularization; hyperplasia; intraluminal angioplasty; laboratory rabbit; laboratory rat; mitogen activated protein kinase; muscle cells; swine; tissue /cell culture; transfection /expression vector; vascular smooth muscle

Major clinical problems caused by unchecked vascular smooth muscle (VSM) proliferation and migration (intimal hyperplasia) can occur following surgical by-pass leading to vein-graft failure or restenosis following arterial angioplasty. Several agents present in serum have been shown to stimulate mitogenesis via dissociated heterotrimeric G proteins (Gq and Gi) appear to be critically involved in the pathological process of intimal hyperplasia since several serum mitogens, such as angiotensin II and thrombin, can activate cellular receptors that can couple to Gq and/or Gi. Peptide inhibitors targeted specifically against Galphaq (GqI) and Gbetagamma (BetaARKct) signaling have recently been developed and thus, molecular inhibitors of Galphaq and Gbetagamma-mediated signaling may represent a novel therapeutic approach to limit vein-graft failure and arterial restenosis after angioplasty. The Overall Hypothesis of this proposal is that G proteins are a critical regulator of VSM cellular proliferation and migration, especially in injured vessels. Furthermore, the targeted inhibition of intracellular G protein signaling, including Gbetagamma inhibition, will lead to a broader understanding of the molecular mechanisms involved in these processes, which may lead to novel molecular therapies to limit the pathological process of intimal hyperplasia. Several in vitro and in vivo model systems are available to test this hypothesis including a rabbit vein-graft model and a porcine coronary artery angioplasty and stent restenosis model. To delivery the GqI and betaArkct peptides to VSM cells in culture and in vivo to the vascular wall, we will utilize adenoviral vectors. The associated Specific Aims are: (1) To identify the specific role of Galphaq and Gbetagamma in the activation of cellular migration and proliferation in VSM cells using in vitro model systems; (2) To determine if in vivo inhibition of Galphaq or Gbetagamma signaling in VSM via adenoviral-mediated gene transfer will limit intimal hyperplasia during vein-graft failure; (3) To determine if adenoviral-mediated delivery of the GqI and betaArkct transgenes to the arterial wall will prevent restenosis is clinically relevant animal models; (4) To evaluate gene transfer to VSM cells within the vessel wall using "advanced" adenovirus and AAV vectors in attempts to more efficiently deliver the peptide inhibitors of Galphaq and Gbetagamma.

在手术旁通后，可能会出现由未检查的血管平滑肌（VSM）增殖和迁移（内膜增生）引起的主要临床问题，导致动脉血管成形术后静脉移动衰竭或再狭窄。 血清中存在的几种药物已被证明可以通过分离的异三聚体G蛋白（GQ和GI）刺激有丝分裂发生，因为几种血管菌素II和血栓蛋白等几种血管蛋白可以激活可以激活夫妇到GQ和/或GI和/或gi。 最近已经开发了针对Galphaq（GQI）和Gbetagamma（BetaArkCT）信号传导的肽抑制剂，因此，Galphaq和Gbetagamma介导的信号传导的分子抑制剂可能代表了一种新型的治疗方法，可以限制静静脉 - 移植衰竭和静脉外的静脉衰减症。 该提议的总体假设是G蛋白是VSM细胞增殖和迁移的关键调节剂，尤其是在受伤的血管中。此外，靶向抑制细胞内G蛋白信号传导（包括gbetagamma抑制）将导致对这些过程所涉及的分子机制的广泛理解，这可能会导致新的分子疗法以限制内膜增生的病理过程。 几种体外和体内模型系统可用于检验该假设，包括兔静脉移植模型以及猪冠状动脉血管成形术和支架再狭窄模型。 为了将GQI和Betaarkct肽递送到培养物和体内的VSM细胞中，我们将利用腺病毒载体。 相关的特定目的是：（1）使用体外模型系统确定Galphaq和Gbetagamma在VSM细胞中细胞迁移和增殖激活中的特定作用； （2）确定通过腺病毒介导的基因转移在VSM中对Galphaq或Gbetagamma信号传导的体内抑制是否会限制静脉移植衰竭期间的内膜增生； （3）确定腺病毒介导的GQI和BetaArkct转基因向动脉壁的递送是否会预防再狭窄是临床相关的动物模型； （4）使用“高级”腺病毒和AAV载体评估基因转移到血管壁中的VSM细胞中，以更有效地传递Galphaq和Gbetagamma的肽抑制剂。