PROJECT 2- MOLECULAR & CELLULAR MECHANISMS AAs

项目 2-分子

• 批准号: 7837663
• 负责人: SHINYA SHIBUTANI
• 金额: $ 43.66万
• 依托单位: STATE UNIVERSITY NEW YORK STONY BROOK
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7837663
• 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; Molecular Toxicology; 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.

首席研究员/项目总监（最后、第一、中间）：Grollman, Arthur P 描述：项目 2 马兜铃酸 (AA) 是马兜铃属植物的主要成分，是一种有效的 人类肾毒素和致癌物质。其毒性作用主要针对肾近曲小管，其中 它会引起细胞损伤，导致皮质肾小管间质纤维化，并最终导致慢性肾功能衰竭。这 这种肾病的病理学已明确，但对其生化机制知之甚少。 AA 发挥其肾毒性作用或如何促进器官特异性癌变。我们假设 AA 和/或其代谢物之一通过两种独立且不同的细胞机制发挥作用；即，（一） 肾近曲小管细胞中转运蛋白介导的浓度和代谢，其中毒素诱导 肾小管损伤； (b) 通过与 DMA 反应形成 dA- 和 dG-AA 加合物，从而引发肿瘤变化 尿路上皮细胞。一系列实验模型，包括整体动物、灌注肾、离体肾 近曲小管和细胞培养将用于确定肾脏处理 AA 的机制和 用于 AA 代谢转化的生化途径。我们将利用自然发生的 对 AA 毒性敏感或具有抗性的小鼠品系可识别与相关相关的过程 易感性。离体灌注大鼠肾、从小鼠和人肾中分离出的肾近端小管、 和细胞培养模型，将用于检查 AA 及其代谢物的机制 在近端小管中运输并发挥其毒性作用，重点是定义 肾毒性反应中的线粒体和氧化损伤。参与近曲小管的通路 AA 的反应通过上皮间质转化诱导肾小管间质纤维化，以及 人类输尿管尿路上皮细胞对 AA 的反应，因为它与 DMA 损伤和致癌作用有关，将是 通过基因阵列表达谱和蛋白质组学分析进行研究。 AA-DNA的形成和修复 加合物将在体内用 AA 肾病小鼠模型进行探索，并在体外用分离的小鼠和 人类肾小管和人类输尿管尿路上皮细胞培养，重点是定义机制 与 AA 肾毒性和遗传毒性有关。最后，我们将识别参与激活和激活的基因和酶。 AA 及其代谢物的解毒作用，认为遗传多态性可能会导致某些 通过改变这些细胞的表达或功能，使个体罹患 AA 肾病和/或尿路上皮细胞癌 酶。