Shp2 a Non-Receptor Tyrosine Phosphatase Regulates Nephrin Phosphorylation and Po

Shp2 非受体酪氨酸磷酸酶调节去氧肾上腺素磷酸化和 Po

• 批准号: 8629068
• 负责人: PUNEET GARG
• 金额: $ 33.82万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-01-01 至 2018-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8629068
• 关键词: Actins; Address; Animal Model; Binding; Biochemical; Biopsy Specimen; Cellular biology; Cytoskeleton; Data; Development; Diabetic Nephropathy; Disease Progression; End stage renal failure; Evaluation; Event; Extracellular Domain; Focal Adhesions; Focal Segmental Glomerulosclerosis; Foot Process; Gene Mutation; Generations; Genes; Homeostasis; Infusion procedures; Injury; Integrins; Investigation; Kidney Diseases; Knockout Mice; Knowledge; Ligands; Link; Maintenance; Maps; Modality; Modeling; Molecular; Morphology; Mus; PTK2 gene; Patients; Phenotype; Phosphorylation; Phosphotransferases; Play; Protamine Sulfate; Protein Tyrosine Phosphatase; Proteins; Proteinuria; Regulation; Renal glomerular disease; Research; Role; Signal Transduction; Techniques; Testing; Tyrosine; Tyrosine Phosphorylation; Urine; Urokinase Plasminogen Activator Receptor; base; cancer therapy; cardiovascular risk factor; genetic regulatory protein; in vivo; inhibitor/antagonist; nephrin; novel; podocyte; prevent; protective effect; protein protein interaction; public health relevance; receptor; research study; response to injury; slit diaphragm; src-Family Kinases; therapeutic target

Project Abstract Proteinuric kidney diseases, including diabetic nephropathy are responsible for majority of the patients that develop end stage renal disease. Proteinuria is not only important for progression of kidney disease but is also an important cardiovascular risk factor. Podocytes have been the main focus of investigations in glomerular diseases that present with proteinuria. Recent studies suggest that podocyte foot process effacement is a form of podocyte mobility that is a result of increase in lamellipodia formation. Nephrin ability to regulate podocyte actin dynamics, lamellipodia formation and focal adhesion dynamics in a phosphorylation dependent manner suggests its role in foot process effacement. Hypothesis: Hypothesis: Inhibition of signaling events that increase podocyte mobility or prevent lamellipodia formation will prevent podocyte foot process spreading and loss following injury. Rationale: Increase in Nephrin phosphorylation has been observed following podocyte injury. Our preliminary data suggests that Src kinase dependent Nephrin phosphorylation is regulated by non- receptor tyrosine phosphatase shp2. Furthermore, inhibition of Shp2 prevents podocyte foot process effacement in protamine sulfate model of podocyte injury. Specific Aims: We will test our hypothesis by pursuing the following two specific aims. 1) Define regulation of Nephrin phosphorylation by Shp2. Using biochemical and cell biology techniques we will expand on our preliminary observations and define the protein-protein interaction and functional consequences of Shp2 dependent Nephrin phosphorylation. 2) Characterize signaling mechanism by which integrin activation regulates Nephrin phosphorylation. We will examine the molecular mechanism that result in Nephrin phosphorylation. Our preliminary experiments suggest that Nephrin phosphorylation occurs as a result of integrin activation. 2) Determine the role of Shp2 and Nephrin phosphorylation on podocyte homeostasis in vivo. We will generate an inducible and a non-inducible podocyte specific conditional shp2 knockout mouse. This mouse will enable us to study the role of shp2 during development and following injury. We will also be able to examine the benefit of preventing foot process effacement in podocyte injury models like protamine sulfate specifically in regards to podocyte loss and proteinuria. Using these mice we will also be able to test other models of podocyte injury.

项目摘要 包括糖尿病性肾病在内的蛋白尿肾脏疾病是导致大多数患者 发展末期肾脏疾病。蛋白尿不仅对肾脏疾病的进展很重要，而且也很重要 重要的心血管危险因素。足细胞一直是肾小球研究的主要重点 蛋白尿中存在的疾病。最近的研究表明，足细胞脚步过程的效率是一种形式 足细胞迁移率的增加，这是层状形成的增加。肾素调节足细胞的能力 肌动蛋白动力学，层状形成和局灶性粘附动力学以磷酸化依赖性方式 暗示其在脚步过程中的作用。假设：假设：抑制信号事件 增加足细胞的迁移率或预防片状脂蛋白的形成将防止脚足迹过程扩散和 受伤后的损失。理由：已经观察到肾素磷酸化的增加 受伤。我们的初步数据表明，SRC激酶依赖性肾素磷酸化受非 - 调节 受体酪氨酸磷酸酶SHP2。此外，抑制SHP2可防止足够的足迹 足细胞损伤的硫酸鱼精素模型中的extracement。 具体目的：我们将通过追求以下两个具体目标来检验我们的假设。 1）定义调节 SHP2的肾素磷酸化。使用生化和细胞生物学技术，我们将在我们的 初步观察并定义SHP2的蛋白质蛋白质相互作用和功能后果 依赖性肾素磷酸化。 2）表征整合素激活的信号传导机制 调节肾素磷酸化。我们将检查导致肾素的分子机制 磷酸化。我们的初步实验表明，由于 整合素激活。 2）确定SHP2和肾素磷酸化对足细胞稳态的作用 体内。我们将产生一种可诱导的和不可诱导的Podocyte特异性条件SHP2基因敲除小鼠。 该小鼠将使我们能够研究SHP2在发育和受伤后的作用。我们也会 能够检查在精完全损伤模型中预防脚步过程的好处 硫酸盐专门针对足细胞丢失和蛋白尿。使用这些老鼠，我们还可以测试 其他足细胞损伤模型。