Role of Focal Adhesion Kinase (FAK) in Nephrosis and Nephritis

粘着斑激酶 (FAK) 在肾病和肾炎中的作用

基本信息

批准号：
8724480
负责人：
Shuta Ishibe
金额：
$ 34万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-07-01 至 2015-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8724480
关键词：
Accounting Actins Address Adhesives Affect Alkylating Agents Angiotensin Receptor Angiotensin-Converting Enzyme Inhibitors Basement membrane Blood Proteins Cell Culture Techniques Cell Line Cells Chronic Kidney Failure Complex Cre-LoxP Cytoskeleton DNA Sequence Rearrangement Data Defect Deterioration Development Diabetic Nephropathy Dialysis procedure Disease Progression End stage renal failure Endothelium Enzymes Event Exhibits Expenditure Extracellular Matrix Failure Filtration Focal Adhesion Kinase 1 Focal Adhesions Foot Process Functional disorder Gelatinase A Glomerular basement membrane antibody Glomerulonephritis Goals Hematuria In Vitro Inflammation Injection of therapeutic agent Injury Injury to Kidney Integrins Kidney Kidney Diseases Kidney Failure Kidney Transplantation Knock-out Knockout Mice Mechanics Mediating Mediator of activation protein Medicare Metalloproteases Methods Modeling Molecular Genetics Mus Mutate Nephritis Nephrosis Oryctolagus cuniculus Parietal Patients Play Process Proteins Proteinuria Public Health Recovery Regulation Renal function Renal glomerular disease Research Project Grants Resistance Rodent Model Role Secondary to Severities Signal Transduction Signaling Molecule Site Steroids Streptozocin Syndrome System Testing Therapeutic Therapeutic Agents Tissue Inhibitor of Metalloproteinases United States Urine Wild Type Mouse anti glomerular basement membrane antibody nephritis cell motility glomerular basement membrane in vivo in vivo Model inhibitor/antagonist injured injury and repair insight kinase inhibitor knock-down migration mouse development mouse model nephrogenesis neuronal cell body novel podocyte public health relevance repaired research study response slit diaphragm small hairpin RNA therapeutic target

项目摘要

DESCRIPTION (provided by applicant): Chronic kidney disease (CKD) often leads to irreversible deterioration of renal function and progresses to End Stage Kidney Disease (ESKD). CKD has emerged as a serious public health problem and data obtained from the USRDS reveals that the number of new cases of ESKD in the United States is projected to be 650,000by 2010, with accompanying Medicare expenditures of $28 billion. As glomerular diseases secondary to podocyte dysfunction contribute up to 90% of all ESKD, a detailed molecular and genetic approach to identify mechanisms for podocyte development and repair may give us new insights for developing therapeutic agents and targets. Currently the therapeutic options available to treat glomerular diseases are limited to Angiotensin Receptor Blockers. Angiotensin Converting Enzyme Inhibitor, Steroids, and Alkylating Agents. As many patients treated with these agents still progress to ESKD, this suggests that other mechanisms responsible for injury are likely involved. When podocytes are damaged, the cell body retracts resulting in effacement and subsequently, proteinuria. For effacement to occur, cells must regulate adhesive contacts between the glomerular basement membrane and the extracellular matrix, which is comprised of focal adhesions and integrins. The goal of this research project is to define the role of focal adhesion protein, Focal Adhesion Kinase (FAK), a critical regulator of cell movement, in podocyte regulation following injury. Preliminary results demonstrate that FAK is highly activated following podocyte injury and a conditional knockout mice lacking podocyte FAK expression appear resistant to injury in murine modes of nephrotic and nephritic syndromes. The aim of the current proposal is to assess the functional relevance of FAK activation by inducing podocyte injury in-vivo and by knocking down podocyte FAK expression with shRNA, and mutating critical FAK regulatory sites in-vitro (Specific Aim 1). As a novel specific FAK inhibitor (Novartis) is currently available, the functional response to FAK inhibition in vivo will be tested before and after podocyte injury in mice using this compound (Specific Aim 2). Finally, to address FAK's role in its regulation of downstream signaling molecules such as inducing matrix metalloproteinase 2 (MMP-2) activity, mice as well as cell culture models defined in the first two specific aims will be utilized to determine the mechanism inducing injury (Specific Aim 3).

描述（由申请人提供）：慢性肾脏疾病（CKD）通常会导致肾功能不可逆的恶化，并发展为终阶段肾脏疾病（ESKD）。 CKD已成为一个严重的公共卫生问题，从USRDS获得的数据显示，美国ESKD的新案件数量预计为2010年为650,000，而Medicare支出为280亿美元。由于肾小球功能障碍继发的肾小球疾病占所有ESKD的90％，因此一种详细的分子和遗传方法，用于鉴定足细胞发育和修复机制，可以为我们提供开发治疗剂和靶标的新见解。目前，可用于治疗肾小球疾病的治疗选择仅限于血管紧张素受体阻滞剂。血管紧张素转化酶抑制剂，类固醇和烷基化剂。由于许多用这些药物治疗的患者仍会发展为ESKD，这表明可能涉及其他负责伤害的机制。当足细胞受损时，细胞体会缩回导致eq eff，随后会导致蛋白尿。为了发生效率，细胞必须调节肾小球基底膜和细胞外基质之间的粘附接触，该基质由局灶性粘连和整联蛋白组成。该研究项目的目的是定义局部粘附蛋白，局灶性粘附激酶（FAK）的作用，局灶性粘附激酶（FAK）是细胞运动的关键调节剂，在损伤后调节足细胞调节中。初步结果表明，在足细胞损伤后，FAK高度激活，而缺乏足细胞FAK表达的有条件敲除小鼠在肾病和肾素性综合征的鼠模式下似乎对损伤具有抵抗力。当前建议的目的是通过在体内诱导足细胞损伤并用SHRNA击倒Podocyte FAK表达并突变关键的FAK调节位点，以评估FAK激活的功能相关性（特定目标1）。作为一种新型的特异性FAK抑制剂（Novartis），使用该化合物在小鼠的小鼠损伤之前和之后测试了对体内FAK抑制的功能反应（特定的AIM 2）。最后，为了解决FAK在调节下游信号分子（例如诱导基质金属蛋白酶2（MMP-2）活性）中的作用，将利用前两个特定目标中定义的小鼠以及细胞培养模型来确定诱导损伤的机制（特定目标3）。