ILK SIGNAL & CYCLIN DL EXPRESSION POST ARTERIAL INJURY

伊尔克信号

• 批准号: 6726082
• 负责人: MICHAEL A. REIDY
• 金额: $ 26.53万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-04-01 至 2007-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6726082
• 关键词: biological signal transduction; blocking antibody; cadherins; calmodulin dependent protein kinase; cardiovascular injury; carotid artery; cyclins; integrins; laboratory mouse; laboratory rat; transfection; vascular smooth muscle

DESCRIPTION (provided by the applicant): We have recently observed that the integrin-linked signaling pathway (ILK) is active in replicating intimal smooth muscle cells of balloon catheter injured arteries. The focus of this proposal is to determine if the activation of this signaling pathway is responsible for the expression of cyclin Dl and ultimately for the replication of intimal SMC. Experiments are designed to examine the downstream targets of ILK in injured rat arteries and will include the activation of GSK-3, the expression and nuclear location of f3-catenin and the activation of the Lef/B-catenin sites on the cyclin Dl promoter. The next aim will identify the upstream regulators of ILK and experiments are proposed to investigate the importance of integrin activation and the P13K pathway. Blocking antibodies will be used to inhibit B1 and B3 integrin activation in injured arteries and ILK activity measured. Wortmannin will be used to inhibit the P13K pathway in replicating intimal SMC and ILK activity quantitated. The last aim will use mice with the ILK gene flanked by Lox sites and breed them with mice expressing smooth muscle specific Cre mice. The resultant off spring will have ILK deleted in SMC and these mice will be then used for injury studies. Cyclin Dl expression and SMC replication will then be measured at a various times after carotid artery injury. Strategies are proposed to circumvent the possibility that such mice are not viable. Finally an adenovirus expressing dominant negative ILK will be used to transfect intimal SMC and both cyclin Dl expression and intimal SMC replication measured. These studies will allow us to demonstrate the importance of the ILK-signaling pathway for the expression of cyclin Dl and the replication of intimal SMC.

描述（由申请人提供）：我们最近观察到 整合素相关信号通路 (ILK) 在复制内膜平滑过程中具有活性 球囊导管损伤动脉的肌肉细胞。本提案的重点 是为了确定该信号通路的激活是否负责 cyclin D1 的表达并最终促进内膜 SMC 的复制。 实验旨在检查受伤者中 ILK 的下游靶标 大鼠动脉，包括 GSK-3 的激活、表达和 f3-连环蛋白的核定位和 Lef/B-连环蛋白位点的激活 细胞周期蛋白D1启动子。下一个目标是确定上游监管机构 提出ILK和实验来研究整合素的重要性 激活和 P13K 途径。封闭抗体将用于抑制 B1 受损动脉中的 B3 整合素激活和测量的 ILK 活性。 渥曼青霉素将用于抑制内膜 SMC 复制中的 P13K 通路 和ILK活性定量。最后一个目标是使用带有 ILK 基因的小鼠 两侧是 Lox 位点，并与表达平滑肌特异性的小鼠进行繁殖 Cre 小鼠。由此产生的后代将在 SMC 中删除 ILK，这些小鼠 然后将用于伤害研究。 Cyclin D1 表达和 SMC 复制 然后将在颈动脉损伤后的不同时间进行测量。 提出了一些策略来避免此类小鼠不存在的可能性 可行的。最后，表达显性失活 ILK 的腺病毒将用于 转染内膜 SMC 以及细胞周期蛋白 D1 表达和内膜 SMC 复制 测量。这些研究将使我们能够证明 用于细胞周期蛋白 D1 表达和复制的 ILK 信号通路 内膜平滑肌细胞。