SIGNAL TRANSDUCTION IN VASCULAR SMOOTH MUSCLE

血管平滑肌的信号转导

• 批准号: 6110456
• 负责人: GARY L STILES
• 金额: $ 25.99万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-01-01 至 2000-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6110456
• 关键词: CHO cells; G protein; adenosine; adenylate cyclase; biological signal transduction; cell cycle; congestive heart failure; guanylate cyclase; membrane proteins; neurotransmitter receptor; phospholipase C; phosphorylation; receptor coupling; recombinant DNA; tissue /cell culture; transfection; vascular smooth muscle; western blottings

Adenosine receptors (AR) play a major role in maintaining and regulating cardiovascular function. The physiological effects of adenosine range from control of chronotropy and ionotropy to vasoregulation and the modulation of growth and migration of smooth muscle cells. Congestive heart failure is associated with decreased vasoregulatory response to adenosine and small vessel vasospasm. In this project, the investigators will characterize AR mediated transmembrane signalling in vascular smooth muscle cells (VSMC) including a delineation of which AR are present, which effector systems are utilized in signalling, how AR regulate growth and differentiation, how elevated levels of adenosine regulate these receptor systems and how these functions can be modified through creation of mutant ARs with particular emphasis on AZARs. Evidence will also be sought to determine if the newly found A2b and A3AR are present in VSMC since physiological data are consistent with their presence. Studies will be carried out in primary cultures of VSMCs, a transformed VSMC line and in CHO and COS7 cells transfected with wild type or mutant receptors. The process of desensitization will be probed in detail concerning the role of phosphorylation of the receptor, mechanisms by which the cell cycles AR and how mutation of potential regulatory sites on the AR changes the patterns of desensitization. Strong evidence exists that the A1AR can regulate multiple effector systems in VSMCs including adenylylcyclase, phospholipase C and guanylate cyclase. A2AR receptors appear to only stimulate adenylylcyclase. Evidence suggests that activation of A2ARs inhibit and A1ARs stimulate growth and DNA synthesis in VSMC. These pathways will be studied in detail and approaches to selectively differentiate pathways that lead to regulation of muscle tone such as inhibition of adenylylcyclase versus regulation of growth and DNA synthesis which may well be mediated by phospholipase C will be undertaken. Attempts to modulate growth of SMCs through transfection (infection) of mutant receptors such as constitutively active AR or dominant negative AR will be undertaken using high efficiency adenovirus vectors. The overall goal will be to understand AR mediated transmembrane signalling in VSMCs, how it is regulated and how it is possible to manipulate these pathways via recombinant DNA techniques for potential therapeutic benefit.

腺苷受体（AR）在维持和调节中起主要作用 心血管功能。 腺苷的生理作用包括 变时性和离子变性对血管调节和调节的控制 平滑肌细胞的生长和迁移。 充血性心力衰竭 与腺苷和小分子血管调节反应降低有关 血管痉挛。 在这个项目中，研究人员将描述 AR 血管平滑肌细胞 (VSMC) 介导的跨膜信号传导 包括描述哪些 AR 存在、哪些效应器系统存在 AR如何用于信号传导，如何调节生长和分化，如何 腺苷水平升高调节这些受体系统以及这些受体系统如何 可以通过创建具有特定功能的突变 AR 来修改功能 强调AZAR。 还将寻求证据以确定新的 发现 A2b 和 A3AR 存在于 VSMC 中，因为生理数据是 与他们的存在一致。 研究将在小学进行 VSMC、转化的 VSMC 系以及 CHO 和 COS7 细胞中的培养物 用野生型或突变型受体转染。 的过程 将详细探讨脱敏的作用 受体的磷酸化，细胞周期AR和的机制 AR 上潜在调控位点的突变如何改变模式 的脱敏作用。 强有力的证据表明 A1AR 可以调节 VSMC 中的多个效应系统，包括腺苷酸环化酶、磷脂酶 C和鸟苷酸环化酶。 A2AR受体似乎只刺激 腺苷酸环化酶。 有证据表明 A2AR 的激活会抑制和 A1AR 刺激 VSMC 的生长和 DNA 合成。 这些途径将是 详细研究了选择性区分途径的方法 导致肌张力调节，例如抑制腺苷酸环化酶 与生长和 DNA 合成的调节相比，这很可能是由 将进行磷脂酶C。 尝试调节 SMC 的生长 通过突变受体的转染（感染），例如组成型受体 主动 AR 或显性负 AR 将采用高效率进行 腺病毒载体。 总体目标是了解 AR 介导的 VSMC 中的跨膜信号传导及其调控方式 可以通过重组DNA技术操纵这些途径 潜在的治疗益处。