PROJECT 2- MOLECULAR & CELLULAR MECHANISMS AAs

项目 2-分子

• 批准号: 8069936
• 负责人: SHINYA SHIBUTANI
• 金额: $ 43.8万
• 依托单位: STATE UNIVERSITY NEW YORK STONY BROOK
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-05 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8069936
• 关键词: Affect; Animals; Aristolochia; Aristolochic Acids; Attention; Biochemical; Biochemical Pathway; Biochemical Process; Carcinogens; Cell Culture Techniques; Cells; Chronic Kidney Failure; DNA; DNA Adducts; DNA Damage; Drug Metabolic Detoxication; Enzyme Gene; Enzymes; Epithelial; Experimental Models; Fibrosis; Gene Expression; Genes; Genetic Polymorphism; Human; In Vitro; Individual; Injury; Kidney; Kidney Diseases; Malignant Neoplasms; Mediating; Mesenchymal; Metabolic; Metabolism; Mitochondria; Modeling; Molecular; Molecular Profiling; Mouse Strains; Mus; Nephrotoxic; Organ; Pathology; Pathway interactions; Plant Components; Predisposition; Principal Investigator; Process; Proteomics; Proximal Kidney Tubules; Rattus; Reaction; Relative (related person); Renal tubule structure; Resistance; Rodent; Role; Toxic effect; Toxin; Tubular formation; Urinary tract; Urothelial Cell; adduct; aristolochic acid II; carcinogenesis; cell injury; epithelial to mesenchymal transition; genotoxicity; in vivo; kidney cell; mouse model; neoplastic; programs; repaired; response

Principal Investigator/Program Director (Last, First, Middle): Grollman, Arthur P DESCRIPTION: PROJECT 2 Aristolochic acid (AA), the principal component of plants belonging to the genus Aristolochiae, is a potent human nephrotoxin and carcinogen. Its toxic effects are targeted primarily to the renal proximal tubule where it causes cellular injury leading to cortical tubulointerstitial fibrosis, and ultimately, chronic renal failure. The pathology of this nephropathy is well defined but little is known about the biochemical mechanisms by which AA exerts its nephrotoxic action or how it promotes organ specific carcinogenesis. We hypothesize that AA and/or one of its metabolites act through two separate and distinct cellular mechanisms; namely, (a) transporter-mediated concentration and metabolism in renal proximal tubule cells, where the toxin induces tubular injury; and (b) by reacting with DMAto form dA- and dG-AA adducts that initiate neoplastic change in urothelial cells. A spectrum of experimental models including whole animal, perfused kidney, isolated renal proximal tubules, and cell culture will be used to identify mechanisms for renal handling of AA and the biochemical pathways used for AA metabolic transformation(s). We will take advantage of naturally occurring mouse strains that are sensitive or resistant to AA toxicity to identify processes associated with relative susceptibility. Isolated perfused rat kidneys, renal proximal tubules isolated from mouse and human kidney, and cell culture models, will be used to examine mechanisms by which AA and its metabolites are transported by, and exert their toxic effects, in the proximal tubule, with a focus on defining the roles of mitochondria and oxidative injury in the nephrotoxic response. Pathways involved in the proximal tubule response to AA in induction of tubulointerstitial fibrosis through epithelial-to-mesenchymal transition, and in the human ureteric urothelial cell response to AA as it relates to DMA damage and carcinogenesis, will be studied by gene array expression profiling and proteomics analysis. The formation and repair of AA-DNA adducts will be explored in vivo with mouse models of AA nephropathy, and in vitro with isolated mouse and human tubules, and human ureteric urothelial cell cultures, with an emphasis on defining mechanisms related to AA nephro- and genotoxicity. Finally, we will identify genes and enzymes involved in activation and detoxification of AA and its metabolites, with the thought that genetic polymorphisms may predispose certain individuals to AA nephropathy and/or urothelial cell cancer by altering the expression or function of these enzymes.

首席调查员/计划主任（最后，第一，中间）：grollman，Arthur P 描述：项目2 亚里士胆胆酸（AA）是属于Aristolochiae属的植物的主要成分，是一种有效的 人肾毒素和致癌物。它的毒性作用主要针对肾近端小管 它会导致细胞损伤导致皮质肾小管间质纤维化，并最终导致慢性肾衰竭。这 这种肾病的病理是很好的定义，但对生化机制知之甚少 AA发挥其肾毒性作用或如何促进器官特定的癌变。我们假设AA 和/或它的一种代谢物通过两个独立和不同的细胞机制作用；即（a） 肾近端小管细胞中转运蛋白介导的浓度和代谢，其中毒素诱导 管状损伤； （b）通过与da-和dg-aa加合物反应，启动肿瘤变化 尿路上皮细胞。一系列实验模型，包括全动物，灌注肾脏，孤立的肾脏 近端小管和细胞培养将用于识别肾脏处理和肾脏的机制 用于AA代谢转化的生化途径（S）。我们将利用自然发生的优势 对AA毒性敏感或抗性的小鼠菌株，以识别与相对相关的过程 敏感性。孤立的灌注大鼠肾脏，从小鼠和人肾脏分离的肾近端小管， 和细胞培养模型，将用于检查AA及其代谢产物的机制 通过近端小管运输并发挥其有毒作用，重点是定义 线粒体和肾毒性反应中的氧化损伤。近端小管所涉及的途径 通过上皮到间质转变诱导拟南芥纤维化的AA的反应 与DMA损伤和致癌作用有关的人类输尿管尿路上皮细胞反应将是 通过基因阵列表达分析和蛋白质组学分析研究。 AA-DNA的形成和修复 与AA肾病的小鼠模型在体内将在体内探索加合物，并在体外与孤立的小鼠和 人小管和人类输尿管尿路上皮细胞培养物，重点是定义机制 与AA肾脏和遗传毒性有关。最后，我们将确定与激活有关的基因和酶 AA及其代谢产物的解毒，认为遗传多态性可能会使某些 通过改变这些人的表达或功能，患有AA肾病和/或尿路上皮细胞癌 酶。