Cell Cycle Control of Restenosis by apoE

apoE 对再狭窄的细胞周期控制

• 批准号: 8492133
• 负责人: Richard Assoian
• 金额: $ 48.88万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8492133
• 关键词: Accounting; Agonist; Alleles; Angioplasty; Animals; Aorta; Apolipoprotein E; Arterial Fatty Streak; Arterial Injury; Arteries; Atherosclerosis; Atomic Force Microscopy; Blood Vessels; Cardiovascular Diseases; Cell Cycle; Cell Cycle Regulation; Cellular biology; Cholesterol; Collaborations; Complication; Consultations; Cyclin D1; Cyclins; Data; Development; Down-Regulation; Epoprostenol; Etiology; Event; Extracellular Matrix; Gene Expression; Gene Proteins; Hepatic; High Density Lipoproteins; Human; Injury; Knockout Mice; Lipid Binding; Measurement; Measures; Mechanics; Mediating; Messenger RNA; Mitogens; Molecular; Molecular Biology; Mus; Mutant Strains Mice; Partner in relationship; Pathway interactions; Phenocopy; Physiological; Plasma; Play; Process; Proteins; Relative (related person); Repression; Research; Resistance; Role; Signal Pathway; Signal Transduction; Site; Smooth Muscle Myocytes; Stimulation of Cell Proliferation; Testing; Therapeutic Intervention; Tissues; Transgenic Mice; arterial stiffness; base; cell injury; cyclin-dependent kinase inhibitor 1B; femoral artery; in vivo; inhibitor/antagonist; injured; mouse model; neointima formation; novel; prevent; protective effect; research study; response; response to injury; restenosis; ubiquitin-protein ligase; vascular smooth muscle cell proliferation

Project 4. ApoE protects against atherosclerosis and restenosis, in part, by reducing plasma cholesterol through hepatic clearance of remnant lipoproteins and HDL. However, newer data indicate that ApoE also protects against atherosclerosis and restenosis independently of lipid-binding. Inhibition of vascular smooth muscle cell (VSMC) proliferation has emerged as a novel, lipid binding-independent effect of ApoE, and this application is focused on elucidating ApoE-dependent anti-mitogenic pathways and testing their relevance in vivo. Our preliminary studies in early passage mouse and human VSMCs have revealed an anti-mitogenic effect of ApoE that Is transduced by the induction of Cox2, synthesis of PGI2, and activation of IP. This ApoE-Cox2-PGl2-IP pathway ultimately controls the levels of the cyclin-dependent kinase Inhibitor, p27'''''^ by regulating the activity of its E3 ligase, SCF^'^P^. Aim 1 will test the importance of the ApoE-Cox2-PGl2-IP pathway in vivo by assessing the response to fine wire arterial injury after deletion and enforced expression of ApoE, Cox2, and IP in the mouse. Aim 2 will test the Importance of Skp2 and p27 as cell cycle targets of ApoE by comparing the response to fine-wire vascular injury In mice lacking Skp2 or p27, or expressing a Skp2-resistant allele of p27 (T187A). Finally, Aim 3 extends preliminary studies showing that the physiological compliance of the mouse aorta and femoral artery, In itself, prevents cell cycling through its inhibitory effect on the mitogen-dependent induction of cyclin D1 mRNA. Thus, the proliferation of VSMCs seen after vascular injury, and the exaggerated proliferation seen after Injury in ApoE-null mice must overcome this control by stiffening the VSMC matrix or by activating a signaling pathway that can induce cyclin D1 in the absence of arterial stiffening. We will directly test these possibilities using milliprobe Indentation or atomic force microscopy to measure homeostatic vessel compliance as well as localized changes in compliance occurring at sites of Injury In the mouse. Related studies will extend preliminary data Indicating that arterial compliance is controlled by the ApoE-Cox2-PGl2-IP pathway and determine the Importance of cyclin D1 in the VSMC injury response in vivo. Together, our studies will characterize new protective effects of ApoE and validate their importance to VSMC proliferation and restenosis in vivo. Our proposed studies closely interact with the Cox2 focus of Project 1 and the focus on mechanical control of cardiovascular disease in Project 5.

项目 4. ApoE 部分通过降低血浆胆固醇来预防动脉粥样硬化和再狭窄 通过肝脏清除残余脂蛋白和高密度脂蛋白。然而，最新数据表明 ApoE 还 防止动脉粥样硬化和再狭窄，与脂质结合无关。抑制血管平滑 肌肉细胞 (VSMC) 增殖已成为 ApoE 的一种新颖的、不依赖于脂质结合的效应，并且这种效应 该应用的重点是阐明 ApoE 依赖性抗有丝分裂途径并测试其在 体内。我们对早期传代小鼠和人类 VSMC 的初步研究揭示了一种抗有丝分裂 ApoE 的作用是通过 Cox2 的诱导、PGI2 的合成和 IP 的激活来转导的。这 ApoE-Cox2-PGl2-IP 途径最终控制细胞周期蛋白依赖性激酶抑制剂 p27'''''^ 的水平 调节其 E3 连接酶 SCF^'^P^ 的活性。目标 1 将测试 ApoE-Cox2-PGl2-IP 的重要性 通过评估删除和强制表达后对细线动脉损伤的反应来评估体内途径 小鼠中的 ApoE、Cox2 和 IP。目标 2 将测试 Skp2 和 p27 作为细胞周期靶标的重要性 ApoE 通过比较缺乏 Skp2 或 p27 或表达 a 的小鼠对细丝血管损伤的反应 p27 (T187A) 的 Skp2 抗性等位基因。最后，目标 3 扩展了初步研究，表明 小鼠主动脉和股动脉的生理顺应性本身就阻止了细胞通过其循环 对细胞周期蛋白 D1 mRNA 的有丝分裂原依赖性诱导的抑制作用。因此，VSMCs的增殖 血管损伤后可见，并且 ApoE 缺失小鼠损伤后观察到的过度增殖必须 通过强化 VSMC 基质或激活信号通路来克服这种控制 没有动脉硬化时的细胞周期蛋白 D1。我们将使用毫探针压痕或原子力显微镜直接测试这些可能性，以测量稳态血管顺应性以及小鼠损伤部位发生的顺应性局部变化。相关研究将扩展初步数据表明动脉顺应性受ApoE-Cox2-PGl2-IP通路控制，并确定cyclin D1在体内VSMC损伤反应中的重要性。我们的研究将共同​​描述 ApoE 的新保护作用，并验证它们对体内 VSMC 增殖和再狭窄的重要性。我们提出的研究与项目 1 的 Cox2 重点和项目 5 的心血管疾病机械控制重点密切相关。