KIDNEY INJURY MOLECULE-1 IN EPITHELIAL REPAIR

肾损伤分子 1 在上皮修复中的作用

• 批准号: 7237837
• 负责人: JOSEPH VINCENT BONVENTRE
• 金额: $ 37.45万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-06-01 至 2010-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7237837
• 关键词: Acute; Acute Kidney Failure; Acute Kidney Tubular Necrosis; Address; Adherens Junction; Adhesions; Adhesives; Agar; Anchorage-Independent Growth; Area; Brain; Cell Adhesion; Cell Aggregation; Cell Communication; Cell Count; Cell Differentiation process; Cell Polarity; Cell membrane; Cell physiology; Cell-Matrix Junction; Cells; Chinese Hamster Ovary Cell; Chronic Kidney Failure; Cisplatin; Cleaved cell; Collagen; E-Cadherin; ERM protein; Epithelial; Epithelial Cells; Epithelium; Event; Extracellular Domain; Extracellular Matrix; Fibronectins; Fibrosis; Functional disorder; Gene Transfer; Genes; Genomics; Goals; Growth; Hand; Heart; Human; Human Cloning; Immunoglobulins; In Vitro; Injury; Integrins; Invasive; Ischemia; Kidney; LLC-PK1 Cells; Lead; Letters; Link; Localized; MDCK cell; Malignant - descriptor; Mechanics; Mediating; Membrane Glycoproteins; Mesenchymal; Mitogen-Activated Protein Kinases; Mitogens; Modeling; Modification; Monoclonal Antibodies; Morbidity - disease rate; Morphology; Mucins; Mus; N-Cadherin; Nuclear; Numbers; Organ; Pathway interactions; Patients; Permeability; Pharmacologic Substance; Phenotype; Phosphotransferases; Polycystic Kidney Diseases; Process; Production; Protein Family; Protein Overexpression; Proteins; RGD (sequence); Rattus; Regulation; Renal Cell Carcinoma; Research Personnel; Rodent; Role; Signal Transduction; Snails; Stress Fibers; Structure; Testing; Tight Junctions; Transcriptional Regulation; Transforming Growth Factor beta; Tubular formation; Type I Epithelial Receptor Cell; Tyrosine Phosphorylation; Urine; Work; adhesion process; cell dedifferentiation; cell motility; epithelial to mesenchymal transition; extracellular; human disease; injury and repair; insight; integrin-linked kinase; interstitial; intracellular protein transport; kidney epithelial cell; migration; mortality; mutant; polyclonal antibody; programs; protein localization location; rat KIM-1 protein; repaired; representational difference analysis; research study; restoration; rho; rho GTP-Binding Proteins; sialylation; therapeutic target; transcription factor; wound

DESCRIPTION (provided by applicant): Kidney injury molecule-1 (KIM-1) is strongly upregulated in proximal tubular epithelial cells in various states characterized by epithelial cell dedifferentiation: ischemia, toxic renal injury, polycystic kidney disease and renal cell carcinoma. It is upregulated more than any other known protein with renal injury. KIM-1 protein is a Type I cell membrane glycoprotein containing extracellular immunoglobulin-like and mucin domains suggesting functional roles in cell-cell and/or cell-matrix interactions. We have cloned, and generated monoclonal and polyclonal antibodies to, the human, mouse and rat KIM-1. KIM-1 is upregulated in vitro in MDCK cells adjacent to a mechanical "wound". The ectodomain of KIM-1 is cleaved and found in urine of patients with acute tubular necrosis or renal cell carcinoma. Gene transfer of KIM-1 in LLC-PK1 epithelial cells results in an increased number of cells with mesenchymal morphology and confers growth in soft agar. The goal of this proposal is to characterize the functional role of KIM-1 during the processes of adhesion and differentiation of epithelial cells. We hypothesize that KIM-1 reduces cell-cell and increases cell-matrix adhesion and potentiates epithelial to mesenchymal transition. In Specific Aim 1, we will characterize the effects of KIM-1 on cell-cell and cell-matrix interactions of epithelial cells. In pilot experiments, we found decreased cell-cell aggregation of LLC-PK1 cells expressing KIM-1. CHO cells overexpressing KIM-1 "scatter" on various matrix substrates. KIM-1 expression results in loss of E-cadherin and increased cell-matrix adhesion. The roles of ERM proteins, integrins, Rho, integrin-linked and mitogen-activated protein (MAP) kinases as effectors of KIM-1 actions on cell-cell and cell-matrix adhesion will be explored. The contributions of the extracellular and intracellular domains of KIM-1 and tyrosine phosphorylation of the intracellular domain of KIM-1 in KIM-l's effects on adhesion will be evaluated. In the second specific aim, we will analyze the role of KIM-1 in the processes associated with dedifferentiation of renal tubular epithelial cells that are important for repair of the epithelium with both positive and potentially negative consequences. We will explore the role of TGF-beta in potentiating the effects of KIM-1 on cell differentiation and epithelial to mesenchymal transition. A mechanism is proposed that implicates ERM proteins, Rho, integrin-linked and MAP family kinases as well as a-catenin and the transcription factor Snail, in the effect of KIM-1 on the differentiation status of the renal epithelial cell. KIM-1-expressing proximal tubules localize in areas of the kidney with increased fibrosis in a model of polycystic kidney disease and we will explore if there is an effect of KIM-1 expression on matrix protein production and extracellular matrix remodeling. Understanding the function of KIM-1 will provide important insight into the role of this protein in injury and repair processes of the kidney, and may identify KIM-1 as an important therapeutic target not only for acute and chronic renal disease but also for malignant transformation of the epithelial cell.

描述（由申请人提供）：肾损伤分子-1（KIM-1）在以上皮细胞去分化为特征的各种状态的近端管状上皮细胞中强烈上调：缺血，毒性肾脏损伤，多囊性肾脏疾病和肾细胞癌。它的上调比其他任何已知的肾脏损伤都要多。 KIM-1蛋白是一种I型细胞膜糖蛋白，含有细胞外免疫球蛋白样和粘蛋白结构域，表明在细胞细胞和/或细胞 - 矩阵相互作用中功能作用。我们已经克隆并产生了与人，小鼠和大鼠KIM-1的单克隆和多克隆抗体。在与机械“伤口”相邻的MDCK细胞中，KIM-1在体外上调。 KIM-1的外域域被裂解并在急性管状坏死或肾细胞癌患者的尿液中发现。 KIM-1在LLC-PK1上皮细胞中的基因转移会导致数量增加具有间充质形态的细胞，并赋予软琼脂的生长。该建议的目的是表征KIM-1在上皮细胞的粘附和分化过程中的功能作用。我们假设KIM-1可以减少细胞细胞并增加细胞矩阵粘附，并增强上皮到间质转变。在特定的目标1中，我们将表征KIM-1对上皮细胞细胞细胞和细胞矩阵相互作用的影响。在试验实验中，我们发现表达KIM-1的LLC-PK1细胞的细胞细胞聚集减少。 CHO细胞过表达KIM-1在各种基质底物上的“散射”。 KIM-1表达会导致E-钙粘着蛋白的丧失和细胞矩阵粘附增加。 ERM蛋白，整联蛋白，RHO，整联蛋白连接和有丝分裂原激活蛋白（MAP）激酶作为KIM-1作用对细胞细胞和细胞 - 矩阵粘附的效应子的作用。将评估KIM-1的KIM-1和酪氨酸磷酸化的细胞外和细胞内结构域在KIM-1对KIM-1对粘附作用中的细胞内磷酸化的贡献。在第二个特定目的中，我们将分析KIM-1在与肾小管上皮细胞的去分化相关的过程中的作用，这些细胞对于修复上皮和潜在负面后果很重要。我们将探讨TGF-beta在增强KIM-1对细胞分化和上皮对间质转变的影响方面的作用。提出了一种机制，该机制暗示了ERM蛋白，RHO，整联蛋白连接和MAP家族激酶以及A-catenin和转录因子蜗牛对KIM-1对肾上皮细胞分化状态的影响。在多囊肾脏疾病模型中，KIM-1表达的近端小管定位在肾脏区域，纤维化增加，我们将探索KIM-1表达对基质蛋白产生和细胞外基质重塑的影响。了解KIM-1的功能将为该蛋白在肾脏的损伤和修复过程中的作用提供重要的见解，并且可以将KIM-1识别为重要的治疗靶标，不仅是急性和慢性肾脏疾病的重要治疗靶标上皮细胞。