Cell Cycle Control of Restenosis by apoE

apoE 对再狭窄的细胞周期控制

• 批准号: 7796925
• 负责人: Richard Assoian
• 金额: $ 50.02万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7796925
• 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; 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 and Cellular 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防止动脉粥样硬化和再狭窄 通过肝脏清除残余脂蛋白和HDL。然而，新的数据表明，ApoE也 不依赖于脂质结合而防止动脉粥样硬化和再狭窄。血管平滑肌抑制 肌肉细胞（VSMC）增殖已成为ApoE的一种新的、不依赖于脂质结合的效应， 本申请集中于阐明ApoE依赖性抗有丝分裂途径并测试它们在 vivo.我们在早期传代小鼠和人VSMCs中的初步研究显示， 通过Cox2的诱导、PGI2的合成和IP的活化转导的ApoE的作用。这 ApoE-Cox2-PG12-IP通路通过以下方式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SCF11P12的活性。目的1将测试ApoE-Cox2-PG12-IP的重要性 通过评估删除和增强表达后对细钢丝动脉损伤的反应， ApoE、Cox2和IP在小鼠中的表达。目的2将测试Skp2和p27作为细胞周期靶点的重要性。 在缺乏Skp2或p27或表达ApoE基因的小鼠中， p27的Skp2抗性等位基因（T187A）。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阻止了细胞通过其 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，血管平滑肌细胞的增殖 在血管损伤后观察到的过度增殖，以及在ApoE缺失小鼠中损伤后观察到的过度增殖， 通过硬化VSMC基质或通过激活可以诱导VSMC增殖的信号通路来克服这种控制。 cyclin D1在动脉硬化的情况下。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。相关的研究将扩展表明动脉顺应性受ApoE-Cox2-PG12-IP通路控制的初步数据，并确定cyclin D1在体内VSMC损伤反应中的重要性。总之，我们的研究将表征ApoE的新保护作用，并验证其对体内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的心血管疾病的机械控制重点密切相关。