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

• 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
• 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

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）通常导致肾功能不可逆恶化，并进展为终末期肾脏疾病（ESKD）。CKD已经成为一个严重的公共卫生问题，从USRDS获得的数据显示，到2010年，美国ESKD的新病例数预计将达到650，000例，伴随的医疗保险支出为280亿美元。由于继发于足细胞功能障碍的肾小球疾病占所有ESKD的90%，因此详细的分子和遗传方法来确定足细胞发育和修复的机制可能会为我们开发治疗药物和靶点提供新的见解。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、类固醇和烷化剂。由于许多接受这些药物治疗的患者仍进展为ESKD，这表明可能涉及导致损伤的其他机制。当足细胞受损时，细胞体收缩，导致消失，随后出现蛋白尿。为了发生消失，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，其由粘着斑和整合素组成。本研究项目的目的是确定粘着斑蛋白，粘着斑激酶（FAK），细胞运动的关键调节因子，在足细胞损伤后的调节中的作用。初步结果表明，FAK是高度活化后足细胞损伤和条件敲除小鼠缺乏足细胞FAK表达出现抗损伤的肾病和肾炎综合征的小鼠模式。本提案的目的是通过在体内诱导足细胞损伤和通过用shRNA敲低足细胞FAK表达以及在体外突变关键FAK调控位点来评估FAK激活的功能相关性（具体目的1）。由于目前可获得一种新型特异性FAK抑制剂（Novartis），因此将在小鼠足细胞损伤前后使用该化合物测试体内对FAK抑制的功能反应（特异性目标2）。最后，为了阐明FAK在其调节下游信号分子（如诱导基质金属蛋白酶2（MMP-2）活性）中的作用，将利用小鼠以及前两个特定目标中定义的细胞培养模型来确定诱导损伤的机制（特定目标3）。