Poldip2: structural and functional implications for vascular disease

Poldip2：对血管疾病的结构和功能影响

• 批准号: 9271231
• 负责人: Kathy K Griendling
• 金额: $ 31.68万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-13 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9271231
• 关键词: Acute; Affect; Alpha Cell; Aneurysm; Animals; Area; Arteries; Atherosclerosis; Autophagocytosis; Binding; Binding Proteins; Biochemistry; Biological Assay; Biology; Birth; Blood Vessels; C-terminal; Cardiovascular Diseases; Cause of Death; Cell Cycle Proteins; Cell Cycle Regulation; Cell Nucleus; Cell Proliferation; Cell physiology; Cells; Cellular biology; Chronic; Collagen; Contracts; Cultured Cells; Cytoplasm; Cytoskeletal Modeling; DNA Damage; Extracellular Matrix; Focal Adhesions; Goals; Growth; Homozygote; Hydrogels; Hydrogen Peroxide; Impairment; Inflammation; Injury; Ischemia; Laboratories; Lesion; Mediating; Mitochondria; Mitotic spindle; Modality; Mus; Muscle Cells; Muscle Proteins; NADPH Oxidase; Oxidases; Peptides; Phase; Phenotype; Physiological; Physiology; Play; Polymerase; Production; Protein Analysis; Proteins; Reactive Oxygen Species; Reagent; Regulation; Role; Site; Smooth Muscle; Smooth Muscle Myocytes; Structure; TNFSF5 gene; Tail; Testing; Therapeutic; Translating; United States; Vascular Diseases; Vascular Smooth Muscle; Vascular remodeling; Work; arterial stiffness; carcinoembryonic antigen-related cell adhesion molecules; cardiovascular risk factor; design; experimental study; extracellular; femoral artery; healing; migration; mutant; novel; novel therapeutic intervention; novel therapeutics; protein function; response; vascular inflammation

PROJECT SUMMARY Reactive oxygen species derived from NADPH oxidases and inflammation have been implicated in most vascular diseases, but also play a role in normal physiology and vascular healing via regulation of smooth muscle cell (SMC) proliferation and matrix secretion. Our laboratory has recently discovered a novel p22phox binding protein, polymerase delta interacting protein-2 (Poldip2), which regulates Nox4, polymerase-δ and cell cycle proteins and has important roles in growth, migration and collagen secretion. In this proposal, we test the hypothesis that the multiple roles of Poldip2 in vascular cells are a function of its binding partners, and that these functions can be specifically targeted using small interfering peptides. In Aim 1, we plan to characterize the physical interaction between Poldip2 and p22phox with the goal of defining peptides that can be used to specifically disrupt this interaction therapeutically. Truncation mutants and pulldown assays will be used to define interacting regions and to design peptides for cell and animal delivery. Aim 2 is devoted to using these peptides to determine the mechanism by which Poldip2 regulates extracellular collagen levels. We will test the hypothesis that depletion of Poldip2 reduces autophagy, thus enhancing collagen secretion. In Aim 3, we will determine the specific consequence of smooth muscle Poldip2-mediated matrix regulation on arterial stiffness and aneurysm formation using Poldipflox/flox animals crossed with smooth muscle-specific Cre mice in both chronic and acute settings, as well as hydrogel- or microparticle-mediated delivery of peptide decoys. Together, the experiments described in this proposal will not only extend our understanding of the vascular biology of Poldip2, but will allow us to take the first step towards translating our findings into a therapeutic modality potentially useful for treating vascular disease.

项目摘要 来源于NADPH氧化酶和炎症的活性氧与大多数肿瘤的发生有关。 血管疾病，但也发挥作用，在正常生理和血管愈合通过调节顺利 肌细胞（SMC）增殖和基质分泌。我们的实验室最近发现了一种新的p22 phox 结合蛋白，聚合酶δ相互作用蛋白-2（Poldip 2），其调节Nox 4、聚合酶-δ和细胞 在生长、迁移和胶原蛋白分泌中具有重要作用。在本提案中，我们测试 假设Poldip 2在血管细胞中的多重作用是其结合伴侣的功能， 可以使用小干扰肽特异性地靶向这些功能。在目标1中，我们计划描述 Poldip 2和p22 phox之间的物理相互作用，目的是定义可用于 在治疗上破坏这种相互作用。将使用截短突变体和下拉试验来 定义相互作用区域并设计用于细胞和动物递送的肽。目标2致力于使用这些 肽，以确定Poldip 2调节细胞外胶原蛋白水平的机制。我们将测试 假设Poldip 2的缺失减少了自噬，从而增强了胶原蛋白分泌。在目标3中，我们 确定平滑肌Poldip 2介导的基质调节对动脉僵硬度的具体结果 和使用Poldipflox/flox动物与平滑肌特异性Cre小鼠杂交的动脉瘤形成， 慢性和急性环境，以及水凝胶或微粒介导的肽诱饵递送。 总之，本提案中描述的实验不仅将扩展我们对血管的理解， Poldip 2的生物学，但将使我们迈出第一步，将我们的发现转化为治疗 可能用于治疗血管疾病的方法。