Technology and Endothelial Biology of Kidney Injury Molecule-1

肾损伤分子1的技术和内皮生物学

基本信息

批准号：
7686935
负责人：
Vishal S. Vaidya
金额：
$ 24.4万
依托单位：
BRIGHAM AND WOMEN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-09-12 至 2011-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7686935
关键词：
Acute Renal Failure with Renal Papillary Necrosis Adult Agar Animals Antibodies Antigens Binding Biological Assay Biological Markers Biology Blood Vessels Boston Candidate Disease Gene Capillary Endothelial Cell Cattle Cell Count Cell Proliferation Cell Survival Cell membrane Cells Characteristics Chemicals Chemotactic Factors Chemotaxis Chimeric Proteins Cisplatin Clear Cell Cleaved cell Clinical Clinical Research Collaborations Cytoplasmic Tail Development Disease Embryo Endothelial Cells Endothelium Environmental Exposure Enzyme-Linked Immunosorbent Assay Epithelial Epithelial Cells Evaluation Extracellular Domain Figs - dietary Gene Expression Growth Growth Factor Hospitals Human Hypoxia Immigration Immunoglobulin Domain Immunoglobulin G In Vitro Individual Injury Integral Membrane Protein Integrins Interleukin-18 Kidney Knock-in Mouse Knock-out Knockout Mice Link Malignant Neoplasms Malignant neoplasm of prostate Measures Mediating Membrane Glycoproteins Mesenchymal Microspheres Modeling Monoclonal Antibodies Morphology Mucins Mus Natural regeneration Nude Mice Oxygen Pathway interactions Patients Pediatric Hospitals Pharmaceutical Preparations Phenotype Process Proteins Proximal Kidney Tubules Rattus Recovery Renal Cell Carcinoma Renal carcinoma Reperfusion Injury Research Rodent Role Sensitivity and Specificity Small Interfering RNA Structure Technology Time Tissues Toxic Environmental Substances Toxic effect Transgenic Mice Tube Tubular formation Tumor Angiogenesis Type I Epithelial Receptor Cell Urine Validation Woman Work Wound Healing angiogenesis base cancer cell cell dedifferentiation cell type environmental chemical extracellular in vivo integrin-linked kinase kidney epithelial cell matrigel migration mortality mouse model neovascularization nephrotoxicity neutrophil osteopontin overexpression paracrine post gamma-globulins pre-clinical promoter protein expression rat KIM-1 protein repaired rho rho GTP-Binding Proteins sulfated glycoprotein 2 tool tubular necrosis tumorigenesis tumorigenic

项目摘要

Kidney Injury Molecule-1 (KIM-1) is a type 1 transmembrane protein that is not detectable in normal kidney tissue but is expressed at very high levels in dedifferentiated proximal tubule epithelial cells in human and rodent kidneys after ischemic or toxic injury. The extracellular domain of KIM-1 is cleaved and can be quantitated, by an ELISA assay, in the urine of rodents and patients with kidney injury or renal cell carcinoma. An important feature common to repair of the tissue after injury (e.g. acute kidney injury, AKI) or growth of cancer (e.g. renal cell carcinoma, RCC) is adequate supply of oxygen through formation of blood vessels. This led us to hypothesize that the function of KIM-1 ectdomain is to activate the endothelial cells by paracrine mechanisms and stimulate angiogenesis. In the preliminary results we found that KIM-1 ectodomain binds to endothelial cells and stimulates migration in a chemotactic and chemoattractant manner. Furthermore, KIM-1 also induced a 2-fold increase in the number of new blood vessels in the in vivo matrigel plug assay indicating its role in neovascularization. The objective of this proposal is to investigate the mechanisms involved in endothelial repair stimulated by KIM-1 following kidney toxicity due to drugs or environmental toxicants. In the first specific aim the expression of soluble KIM-1 will be characterized in relation with other biomarkers involved in endothelial repair process after kidney injury using various in vivo preclinical and clinical models of kidney toxicity. The second aim of this proposal is to investigate the mechanism of angiogenesis by KIM-1 in the context of endothelial cell regeneration by conducting structure function studies to identify the critical domain of KIM-1 responsible for endothelial cell survival, migration, tube formation and new blood vessel growth. We will also investigate if KIM-1 stimulates migration by inducing candidate gene expression further activating Rho and Integrin linked kinase pathways to cause chemotaxis. We will determine the critical role of KIM-1 in angiogenesis and endothelial resurtucturing by "Knock-in" and "Knockout" strategies using Kim-1 transgenic mice that overexpresses KIM-1 selectively on the epithelial cells of the embryonic and adult kidney proximal tubules and investigate if these mice have faster and more effective endothelial regeneration and recovery from kidney toxicity. Consequently, we will also use the KIM-1 knockout mice to determine if they are more susceptible to nephrotoxicity owing to the lack of repair. Since kidney is a major target for toxicity due to chemical annd physical agents, unraveling the mechanisms of how KIM-1 regulates growth of blood vessels would offer treatment paradigms to promote (in AKI) or inhibit (in kidney cancer) angiogenesis to ameliorate or perhaps even cure disorders that are leading causes of mortality today.

肾损伤分子1（KIM-1）是一种1型跨膜蛋白，在正常情况下无法检测到肾脏组织，但在人类的近端小管上表达很高缺血性或有毒损伤后的啮齿动物肾脏。 Kim-1的细胞外域被裂解，可以是通过ELISA测定，在啮齿动物和肾脏损伤或肾细胞患者的尿液中定量癌。受伤后修复组织（例如急性肾脏损伤，AKI）或癌症的生长（例如肾细胞癌，RCC）是通过血液形成的足够的氧气供应船只。这使我们假设Kim-1 ectomain的功能是通过旁分泌机制并刺激血管生成。在初步结果中，我们发现Kim-1 外生域与内皮细胞结合并刺激趋化和趋化剂中的迁移方式。此外，KIM-1还引起体内新血管数量增加了2倍 Matrigel插头测定表明其在新血管中的作用。该提案的目的是研究由肾脏毒性或环境毒性引起的肾脏毒性后的Kim-1。在第一个特定目标中可溶性KIM-1的表达将与其他与内皮相关的生物标志物相关肾损伤后使用各种体内临床前和肾脏毒性临床模型的修复过程。这该提议的第二个目的是研究KIM-1在内皮细胞再生通过进行结构功能研究来确定KIM-1的关键结构域负责内皮细胞存活，迁移，管形成和新血管生长。我们也会研究KIM-1是否通过诱导候选基因表达进一步激活Rho和整合素连接的激酶途径引起趋化性。我们将确定KIM-1在使用KIM-1转基因的“敲入”和“敲除”策略通过“敲入”和“敲除”策略进行血管生成和内皮重塑在胚胎和成年肾脏的上皮细胞上有选择地表达Kim-1的小鼠小管和研究这些小鼠是否具有更快，更有效的内皮再生和恢复来自肾脏毒性。因此，我们还将使用Kim-1敲除小鼠来确定它们是否更多由于缺乏修复，易受肾毒性的影响。由于肾脏是由于化学和物理剂引起的毒性的主要目标，因此 Kim-1如何调节血管生长的机制将提供治疗范例以促进（在 AKI）或抑制（在肾癌中）血管生成可改善甚至可以治愈引起的疾病今天的死亡率。