Molecular Toxicology in Human Kidney Cells

人肾细胞的分子毒理学

• 批准号: 6781240
• 负责人: LAWRENCE H. LASH
• 金额: $ 23.29万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-15 至 2008-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6781240
• 关键词: DNA damage; DNA repair; apoptosis; cell proliferation; chemical conjugate; cytotoxicity; flow cytometry; human tissue; kidney cell; kidney metabolism; mitochondria; mitochondrial membrane; regeneration; renal toxin; renal tubule; tissue /cell culture; toxin metabolism; trichloroethylene; western blottings

DESCRIPTION (provided by applicant): Trichloroethylene (TRI) is a major environmental contaminant, is an established animal carcinogen, and is considered a "probably human carcinogen" by the National Toxicology Program and the International Agency for Research on Cancer. The kidneys are one target organ for TRI and its nephrotoxic and nephrocarcinogenic effects are mediated by metabolites derived from conjugation with glutathione (GSH). Subsequent metabolism to the cysteine conjugate S- (1,2- dichlorovinyl)-L-cysteine (DCVC) generates the penultimate toxic metabolite. It is metabolism of DCVC by either the cysteine conjugate ¿-Iyase or the flavin-containing monooxygenase that generates the ultimate reactive and toxic species. Most of the previous research that has delineated the metabolism and potential modes of action for TRI and DCVC has been performed in rodents or with tissue from rodents. While these studies, some of which have been done by the PI, have provided much useful insight, there are problems in using data obtained from rodents for human health risk assessment. This is particularly true for halogenated solvents such as TRI, because of marked species differences in metabolism, transport, and overall sensitivity to toxicity. Previous studies of ours showed that DCVC can cause both apoptosis or necrosis in primary cultures of human proximal tubular (hPT) cells, depending on concentration and time of exposure. Findings also suggested effects of DCVC on expression of proteins related to stress response and regulation of cell growth. This application uses primary cultures of hPT cells as the experimental model and will investigate the ability of hPT cells exposed to moderately toxic concentrations of DCVC to undergo repair and regeneration, the potential for DCVC to induce cell proliferation by non-genotoxic mechanisms, and the requirement for mitochondrial toxicity in the course of events leading from exposure to toxicity. The application comprises three Specific Aims. Specific Aim 1 addresses the question of whether hPT cells exposed to moderately toxic concentrations of DCVC undergo repair and regeneration. Several markers of repair will be assessed and precise conditions and potential mechanisms by which the repair and regeneration response are induced will be investigated. Specific Aim 2 will address the question of whether DCVC can stimulate uncontrolled proliferation of hPT cells. Effects on cell cycle and cell cycle signaling molecules under various conditions of DCVC exposure will be studied. Finally, Specific Aim 3 will address the question of whether mitochondrial toxicity is sufficient and necessary for DCVC-induced toxicity in hPT cells. Although previous work has shown that mitochondria are early and potently affected intracellular targets of DCVC, it is not known whether mitochondrial toxicity is an obligatory step in the progression of events that occur after DCVC exposure or whether other pathways that are independent of mitochondria can mediate renal cell injury. Achievement of these aims should build on our previous work in human kidney cells and extend it to provide a much more complete understanding of the various and complex ways in which DCVC affects the human kidney

描述（由应用提供）：三氯乙烯（TRI）是一种主要的环境污染物，是一种已建立的动物致癌物，被国家毒理学计划和国际癌症研究机构视为“可能的人类致癌物”。孩子们是三位一体的一个靶器官，其肾毒性和肾癌作用作用是由与谷胱甘肽（GSH）结合的代谢产物介导的。随后对半胱氨酸结合物S-（1,2-二氯苯基）-L-半胱氨酸（DCVC）的代谢会产生倒数第二个有毒代谢物。它是通过半胱氨酸结合»-iyyase或含黄素单加氧酶的DCVC代谢，产生最终的反应性和有毒物种。以前的大多数研究都描述了TRI和DCVC的代谢和潜在的作用模式。尽管这些研究是由PI完成的，但提供了许多有用的见解，但使用从啮齿动物获得的数据进行人体健康风险评估时存在问题。对于卤代溶液（例如TRI）尤其如此，因为新陈代谢，运输和对毒性的总体敏感性的物种差异明显。对我们的先前研究表明，DCVC可能会导致人近端肾小管（HPT）细胞的原发性培养物的细胞凋亡或坏死，这取决于浓度和暴露时间。发现还表明，DCVC对与胁迫反应和细胞生长调节有关的蛋白质表达的影响。应用使用HPT细胞的原发性培养物作为实验模型，并将研究暴露于DCVC中毒性中等毒性浓度进行修复和再生的HPT细胞的能力，DCVC的潜力是通过非生物毒性机制诱导细胞增殖的潜力，以及导致曝光毒性毒性毒性毒性的毒性的需求。该应用程序包括三个特定目标。具体目的1解决了HPT细胞是否暴露于中度毒性的DCVC进行修复和再生的问题。将评估维修的几个标志，并将研究修复和再生反应的精确条件和潜在机制。具体目标2将解决DCVC是否可以刺激HPT细胞不受控制的增殖的问题。在DCVC暴露的各种条件下，将研究对细胞周期和细胞周期信号分子的影响。最后，特定的目标3将解决线粒体毒性是否足够且对于DCVC诱导的HPT细胞毒性所必需的问题。尽管先前的工作表明，线粒体是DCVC的早期和潜在影响的细胞内靶标，但尚不清楚线粒体毒性是否是DCVC暴露后发生的事件的进展，还是其他与线粒体无关的途径可以介导肾细胞损伤的事件的进展。这些目标的实现应建立在我们以前在人类肾细胞中的工作，并将其扩展，以提供对DCVC影响人类肾脏的各种复杂方式的完整理解