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组织中存在，但在人的去分化近端小管上皮细胞中表达水平非常高和啮齿动物肾脏在缺血或中毒性损伤后。 KIM-1 的胞外结构域被切割并且可以通过 ELISA 测定，对啮齿动物和肾损伤或肾细胞患者的尿液进行定量癌。损伤后组织修复所共有的一个重要特征（例如急性肾损伤，AKI）或癌症（例如肾细胞癌，RCC）的生长是通过血液的形成提供足够的氧气船只。这使我们推测 KIM-1 胞外域的功能是通过以下方式激活内皮细胞：旁分泌机制并刺激血管生成。在初步结果中我们发现KIM-1 胞外域与内皮细胞结合并刺激趋化剂和趋化剂的迁移方式。此外，KIM-1 还诱导体内新血管数量增加 2 倍。基质胶塞测定表明其在新血管形成中的作用。该提案的目的是研究刺激内皮修复的机制 KIM-1 因药物或环境毒物导致肾毒性。在第一个具体目标中可溶性 KIM-1 的表达将与内皮细胞相关的其他生物标志物的关系进行表征。使用各种体内肾毒性的临床前和临床模型研究肾损伤后的修复过程。这该提案的第二个目的是研究 KIM-1 在以下情况下的血管生成机制：通过进行结构功能研究以确定 KIM-1 的关键域来促进内皮细胞再生负责内皮细胞的存活、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 来刺激迁移整合素连接激酶途径引起趋化性。我们将确定 KIM-1 在使用 Kim-1 转基因通过“敲入”和“敲除”策略进行血管生成和内皮再生在胚胎和成年肾近端的上皮细胞上选择性过度表达 KIM-1 的小鼠肾小管并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有效。由于缺乏修复而容易产生肾毒性。由于肾脏是化学和物理制剂毒性的主要目标，因此需要解开 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（肾癌）血管生成以改善甚至治愈导致疾病的疾病今天的死亡原因。