Glomerular Capillaries-- Normal and Pathologic

肾小球毛细血管——正常和病理

• 批准号: 8792268
• 负责人: MARILYN Gist FARQUHAR
• 金额: $ 52.4万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-06-05 至 2017-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8792268
• 关键词: Adriamycin PFS; Affect; Animals; Apoptosis; Binding; Biochemical; Biological Assay; Cell Survival; Cells; Complex; Data; Development; Diabetic Nephropathy; Disease; Fluorescence; Focal Adhesions; Focal glomerulosclerosis; Glomerular Capillary; Glomerulonephritis; Goals; Grant; Image; In Situ; Injury; Instruction; Intercellular Junctions; Kidney; Knock-out; Knockout Mice; Life; Ligation; Mediating; Molecular; Mus; Nephrosis; Pathologic; Permeability; Phenotype; Play; Principal Investigator; Process; Progress Reports; Proteins; Proteinuria; Proteomics; Renal glomerular disease; Research; Role; Signal Pathway; Signal Transduction; Testing; Vascular Endothelial Growth Factor Receptor; Vascular Endothelial Growth Factor Receptor-2; Vascular Endothelial Growth Factors; Vinculin; Work; absorption; cellular imaging; db/db mouse; glomerular filtration; insight; mutant; nephrin; novel; podocyte; research study; response; slit diaphragm

During the last 39 years under this grant our research has been directed toward understanding the cellular and molecular mechanisms of glornerular permeability and protein absorption as well as the derangements in these processes that occur in glomerular diseases. The experiments proposed in this application represent a direct continuation of our ongoing work on characterization of slit diaphragms and the role of GIV in podocyte function and survival after podocyte injury. We will focus on three specific aims: Specific Aim # 1 : To characterize and determine the functions of novel, putative slit diaphragm components identified by MS and quantitative organellar proteomics in slit diaphragm fractions. We will verify their junctional localization and determine their binding partners and functions in podocytes. Specific Aim #2: To investigate the role of GIV in mediating Akt signaling and cell survival in passive Heymann nephritis, focal glomerulosclerosis, and diabetic nephropathy. In PAN nephrosis GIV is upregulated, assembles a VEGFR2/GIV/Gal3 complex, and functions to maintain podocyte survival. We will determine if GIV plays a similar role in other diseases associated with podocyte injury. Specific Aim #3: To further investigate the mechanisms and spatial-temporal aspects of GIV's effects on podocytes. Our preliminary data indicate that GIV might function at focal adhesions (FA) and/or at slit diaphragms as it colocalizes with vinculin and VEGFR2 at FA and it binds CD2AP. We will investigate the dynamics ofthe interactions between GIV and VEGFR2 and CD2AP by live cell imaging, use proximity ligation assays and superresolution IF imaging (STORM) to determine where interaction occurs, and test whether GIV depletion or expression of GIV mutants affect FA or junction formation. These studies can be expected to provide novel insights into understanding ofthe cellular and molecular mechanisms of glomerular filtration and glomerular injury and their alterations in glomerular diseases associated with proteinuria. RELEVANCE (See instructions): Podocyte injury is the initiating cause of many glomerular diseases. The studies planned will provide key insights into the signaling neworks that regulate podocyte organization, functions and survival in response to glomerular injury in diseases associated with proteinuria.

在过去的39年中，根据这项赠款，我们的研究旨在理解 Glornerular渗透性和蛋白质吸收的细胞和分子机制以及 这些过程中发生的肾小球疾病中发生的危险。提出的实验 应用代表了我们正在进行的有关缝隙隔膜和表征的工作的直接延续 GIV在足细胞损伤后足细胞功能和存活中的作用。我们将专注于三个具体目标： 特定目的＃1：表征并确定新颖的，推定的缝隙膜片的功能 通过缝隙隔膜级分中MS和定量细胞器蛋白质组学鉴定的成分。我们将 验证其连接定位，并确定其结合伙伴和足细胞中的功能。 特定目的＃2：研究GIV在被动中介导AKT信号传导和细胞存活中的作用 海曼肾炎，局灶性肾小球硬化和糖尿病性肾病。在泛肾病中GIV是 上调，组装vegfr2/giv/gal3复合物，并具有维持足细胞存活的功能。我们 将确定GIV是否在与足细胞损伤相关的其他疾病中起着相似的作用。 特定目的＃3：进一步研究GIV对GIV影响的机制和时空方面 足细胞。我们的初步数据表明，GIV可能在焦点粘附（FA）和/或缝隙中起作用 diaphragms与FA处的Vinculin和Vegfr2共定位，并结合CD2AP。我们将调查 通过活细胞成像，GIV和VEGFR2与CD2AP之间相互作用的动力学，使用接近度 结扎测定和超分辨率如果成像（风暴）以确定发生相互作用的地方，并测试 GIV突变体的GIV耗竭或表达会影响FA或结的形成。 可以期望这些研究为理解细胞和分子的新见解提供新的见解。 肾小球过滤和肾小球损伤的机制及其在肾小球疾病中的改变 与蛋白尿有关。 相关性（请参阅说明）： 足细胞损伤是许多肾小球疾病的起始原因。计划的研究将提供关键 对调节足细胞组织，功能和生存的信号新功能的见解 与蛋白尿相关的疾病中的肾小球损伤。