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: 10299122
• 负责人: JOSEPH VINCENT BONVENTRE
• 金额: $ 54.79万
• 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 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10299122
• 关键词: 3-Dimensional; Acute; Acute Renal Failure with Renal Papillary Necrosis; Affect; Albumins; Algorithms; Anemia; Animal Model; Animals; Antigen Presentation; Architecture; Autophagocytosis; Binding; Binding Sites; Biological Markers; Blood; Canis familiaris; Cardiovascular Diseases; Cell Aging; Cell Cycle Arrest; Cell Line; Cells; Characteristics; Chronic; Chronic Kidney Failure; Cleaved cell; Clinical; Clinical Trials; Complex; Cryoelectron Microscopy; DNA Damage; Development; Diabetic Nephropathy; Diabetic mouse; Drug Targeting; Endocytosis; Epithelial Cells; FRAP1 gene; Family suidae; Fatty Acids; Fibronectins; Fibrosis; Functional disorder; G2/M Arrest; Generations; Genetic Models; Goals; Grant; Heart Hypertrophy; Hemodialysis; Human; Hypertension; Impairment; Inflammation; Inflammatory; Inflammatory Response; Injury; Injury to Kidney; Integrins; Ischemia; Kidney; Kidney Failure; Lead; Link; Lymphoid Tissue; MAP Kinase Gene; Mediating; Mitochondria; Modeling; Molecular; Monoclonal Antibodies; Mucins; Mus; Myofibroblast; Nonesterified Fatty Acids; Nuclear; Organ failure; Organoids; Oxidants; Palmitates; Palmitic Acids; Pathway interactions; Phagocytes; Phagocytosis; Phagosomes; Phenotype; Plasma; Play; Population; Process; Production; Proteins; Proximal Kidney Tubules; Rattus; Regulation; Regulatory T-Lymphocyte; Renal Cell Carcinoma; Reporting; Rodent; Role; Secondary Hypertension; Signal Transduction; Structure; System; Time; Tissues; Toxic effect; Transplantation; Tubular formation; Up-Regulation; Urine; Zebrafish; cell dedifferentiation; cell injury; energy balance; epithelial repair; extracellular; fatty acid oxidation; fatty acid-binding proteins; glomerular filtration; human pluripotent stem cell; inhibitor/antagonist; injured; interstitial; kidney epithelial cell; man; mitochondrial dysfunction; mouse genetics; mouse model; mutant; nephrotoxicity; non-diabetic; novel; oxidized lipid; oxidized low density lipoprotein; phosphatidylserine receptor; polyclonal antibody; pre-clinical; prevent; rat KIM-1 protein; repaired; response; safety study; screening; senescence; side effect; small molecule libraries; therapeutic target; uptake; urinary

Project Summary/ Abstract Kidney Injury Molecule-1 (KIM-1) is the most upregulated protein in proximal tubular epithelial cells in various states characterized by epithelial cell dedifferentiation: ischemia, toxic renal injury, and renal cell carcinoma. We have cloned, generated cells and animals expressing wild-type and mutant KIM-1, and created monoclonal and polyclonal antibodies to, human, mouse, rat, pig, dog, and zebrafish KIM-1. We have reported that the KIM-1 ectodomain is cleaved into the blood and urine of subjects with acute (AKI) and chronic (CKD) kidney injury and is a sensitive and specific kidney injury biomarker to detect kidney injury and predict progression of CKD. KIM-1 has been qualified by the FDA for preclinical and clinical use in kidney safety studies. We have discovered that KIM-1 transforms kidney epithelial cells into semiprofessional phagocytes making it the first nonmyeloid phosphatidylserine receptor. We have described a novel phagocytosis pathway that links autophagy to KIM-1-mediated phagosome maturation and MHC restricted antigen presentation in epithelial cells. We have shown that KIM-1 expression in early AKI is adaptive, but chronic expression leads to CKD with severe fibrosis, secondary hypertension, and cardiac hypertrophy. A mouse lacking the extracellular mucin domain, important for phagocytosis, is protected against development of fibrosis. We have found that KIM-1 mediates uptake of palmitate-bound albumin and recently found an inhibitor of KIM-1- mediated phagocytosis by screening a small molecule library. The inhibitor reduces cell lipotoxicity and fibrosis in a novel mouse model of diabetic kidney disease. The current competing renewal application builds upon and extends our prior findings. Our goal is to further characterize KIM-1-mediated uptake of fatty acid bound albumin (FA-Albumin), and the implications of this uptake for cellular injury and maladaptive repair, including cell senescence leading to profibrotic and proinflammatory responses that ultimately lead to progressive CKD. KIM-1 may be a drug target to prevent and treat CKD. We hypothesize that persistent KIM-1-mediated endocytosis of FA-Albumin and subsequent signaling leads to toxicity. FA-Albumin uptake leads to a mitochondrial dysfunction, DNA damage response (DDR), G2/M arrest, mTOR signaling, TASCC formation, and a prosecretory fibrotic phenotype. In addition KIM-1-FA-Albumin uptake leads to chronic tissue inflammation in part due to tertiary lymphoid tissue development through LTaβ/LTβR signaling. In Specific Aim 1 we will characterize binding of KIM-1 to FA-Albumin and determine the architecture, structural dynamics and molecular basis for FA-Albumin binding to KIM-1. In Specific Aim 2 we will characterize the intracellular consequences of KIM-1 mediated FA-Albumin endocytosis, particularly on mitochondrial function, DNA damage, the DDR, cell cycle arrest and the profibrotic secretome. In Specific Aim 3 we will evaluate the role of KIM-1 mediated FA- Albumin uptake and DDR in LTaβ/LTβR signaling leading to tertiary lymphoid tissue (TLT) formation with consequent pro-inflammatory consequences in animal models of AKI to CKD transition.

项目摘要/摘要 肾损伤分子1（KIM-1）是各种近端肾小管上皮细胞中最新蛋白质的蛋白质 以上皮细胞去分化为特征的状态：缺血，有毒肾损伤和肾细胞癌。 我们已经克隆，产生的细胞和动物表达野生型和突变体Kim-1，并创建了单克隆 和对人，小鼠，大鼠，猪，狗和斑马鱼Kim-1的多克隆抗体。我们报告说 Kim-1外生域被裂解成急性（AKI）和慢性（CKD）肾脏的受试者的血液和尿液 受伤，是一种敏感且特异的肾脏损伤生物标志物，可检测肾脏损伤并预测 CKD。 KIM-1已获得FDA的资格，用于肾脏安全研究中的临床前和临床用途。我们有 发现Kim-1将肾上皮细胞转化为半体型吞噬细胞，使其成为 第一个非甲状腺磷脂酰丝氨酸接收器。我们描述了一种新型的吞噬作用途径 自噬至KIM-1介导的吞噬体成熟和MHC限制性抗原表现 细胞。我们已经表明，早期AKI中的Kim-1表达是自适应的，但慢性表达导致 CKD具有严重的纤维化，继发性高血压和心脏肥大。一只鼠标缺乏 细胞外粘蛋白结构域对吞噬作用很重要，可保护纤维化的发展。我们 发现Kim-1介导了棕榈酸酯结合的白蛋白的摄取，最近发现了Kim-1-的抑制剂 通过筛选一个小分子文库，介导的吞噬作用。抑制剂降低细胞脂毒性和 新型糖尿病肾脏疾病的小鼠模型中的纤维化。当前的竞争续约申请正在建立 并扩展我们的先前发现。我们的目标是进一步表征KIM-1介导的脂肪酸摄取 绑定的白蛋白（FA-Albumin），以及这种摄取对细胞损伤和适应不良修复的影响， 包括细胞感应导致纤维化和促炎反应，最终导致 进步的CKD。 KIM-1可能是预防和治疗CKD的药物靶标。我们假设这一持久 KIM-1介导的FA-醛蛋白的内吞作用和随后的信号传导导致毒性。 FA-Albumin 摄取导致线粒体功能障碍，DNA损伤响应（DDR），G2/m逮捕，mTOR 信号传导，TASCC形成和起诉纤维化表型。另外，Kim-1-FA-Albumin 摄取导致慢性组织注射部分由于第三纪 淋巴组织通过LTAβ/LTβR信号传导发育。在特定目标1中，我们将描述 KIM-1与FA-α蛋白的结合，并确定结构，结构动力学和分子基础 FA-α-与KIM-1结合。在特定目标2中，我们将表征KIM-1的细胞内后果 介导的FA-α-珠蛋白内吞作用，尤其是线粒体功能，DNA损伤，DDR，细胞周期 逮捕和纤维性分泌组。在特定目标3中，我们将评估KIM-1介导的FA-的作用 LTAβ/LTβR信号传导中的白蛋白摄取和DDR导致三级淋巴组织（TLT）形成 随之而来的是AKI到CKD转变的动物模型中的促炎后果。