TGF-BETA 1 RECEPTORS IN RESTENOSIS AND AGING

心血管疾病是老年人发病和死亡的最大原因。 令人信服的证据表明，血管修复细胞中与年龄相关的变化是血管损伤的根本原因。 老年人动脉粥样硬化的发病率增加。人们认为，血管修复的“失败死亡” 在吸烟、高血压和高脂血症损伤后，细胞导致它们在血管壁中积聚。 我们已经证明了人动脉粥样硬化细胞对凋亡因子的功能性抵抗 病变，并在细胞来源于老年Fisher 344大鼠。在延长MERIT的建议中， 奖，我们表明，细胞生物学和微阵列分析的组合可能已经确定了潜在的 因为这个缺陷。人类病变细胞、药物处理细胞和老化啮齿动物动脉的微阵列分析 并且细胞产生了一系列可以赋予细胞凋亡抗性的候选基因。对系统地 通过QPCR、蛋白质印迹和强制过表达分析这些靶点，我们可以确定Bcl-XI 仍然是一个强有力的候选人的致病因素，在年龄相关的抵抗凋亡诱导的Fas 结扎同样，细胞周期蛋白D1和Smurf 2仍然是TGF-β抵抗的可靠候选者。 在MERIT奖的第二个任期，我们继续朝着了解具体的目标前进， 控制血管细胞凋亡抗性的机制。提出三个具体目标：（1）进一步 通过siRNA抑制来恢复功能，并通过强制过表达来诱导这些靶点。 敏感细胞中的表型; 2）使用微阵列和蛋白质组学来定义抗性的整个途径 方法，以找到适合干预的成分; 3）确定调节的效果， 在老化啮齿类动物血管壁中损伤诱导的损伤形成过程中的阻力途径。的 研究已经发现了新的治疗药物，如flavopiridol和picetannol，一种天然的葡萄 产物，其能够调节损伤中对细胞凋亡的抗性。建议的结果 研究将准确界定机制，并确定其他适合干预的步骤。的 这些研究的目的是确定进展性动脉粥样硬化病变的诊断标志物以及 可以通过药物或遗传干预来调节的靶点，以防止 血管壁中的细胞，从而防止心脏病发作和中风。

Cardiovascular disease is by far the largest cause of morbidity and mortality in the elderly population. Compelling evidence indicates that age-related changes in vascular repair cells are a root cause of the increased incidence of atherosclerosis in the elderly. It is thought that a 'failure to die'by vascular repair cells leads to their accumulation in the vessel wall after injury by smoking, hypertension, and hyperlipidemia. We have documented functional resistance to apoptotic factors in cells derived from human atherosclerotic lesions, and in cells derived from aged Fisher 344 rats. In this proposal for an extension of the MERIT Award, we show that a combination of cell biology and microarray profiling has likely identified the underlying cause of this defect. Microarray profiling of human lesion cells, drug-treated cells, and aged rodent arteries and cells produced a list of candidate genes which could confer resistance to apoptosis. In systematic analysis of these targets by QPCR, Western Blot, and forced overexpression, we could determine that Bcl-XI remains a strong candidate for a causative factor in age-related resistance to apoptosis induced by fas ligation. Likewise, cyclin D1 and Smurf2 remain solid candidates as mediators of the resistance to TGF-fi>. In the second term of the MERIT Award, we continue toward the goal of understanding the specific mechanisms that control apoptotic resistance in vascular cells. Three Specific Aims are proposed: 1) further confirm these targets by siRNA inhibition to restore function, and by forced overexpressionto induce the phenotype in sensitive cells; 2) define the entire pathway of resistance using microarrays and proteomic methods to find components that are amenable to intervention; 3) determine the effect of modulating the resistance pathway on the course of injury-induced lesion formation in the aging rodent vessel wall. The studies have already identified novel therapeutic agents, such as flavopiridol and picetannol, a natural grape product, that are capable of modulating resistance to apoptosis in the lesion. The results of the proposed studies will precisely define the mechanism, and identify other steps that are amenable to intervention. The goal of these studies is to identify both diagnositic markers of advancing atherosclerotic lesions as well as targets which can be modulated by Pharmaceuticalsor genetic interventions to prevent the accumulation of cells in the vessel wall, and thereby prevent heart attacks and stroke.