权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Identification and Characterization of Mitochondrial Toxins

线粒体毒素的鉴定和表征

基本信息

批准号：
8127097
负责人：
Lauren P. Wills
金额：
$ 5.13万
依托单位：
MEDICAL UNIVERSITY OF SOUTH CAROLINA
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-08-01 至 2012-09-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8127097
关键词：
Acute Address Adenosine Triphosphate Affect Agriculture Apoptosis Biological Assay Cell Death Cell Respiration Cells Chemical Structure Chemicals Chronic Clinical Computer Simulation Data Databases Development Electron Transport Environmental Exposure Environmental Pollution Evaluation Functional disorder Future Genus Hippocampus Goals Government Agencies Health Human Injury Ions Kidney Lead Libraries Link Methods Mitochondria Modeling Molecular Necrosis Organ Oryctolagus cuniculus Oxidative Phosphorylation Pathway interactions Pesticides Pharmacologic Substance Production Proximal Kidney Tubules Research Respiration Risk Risk Assessment Screening Result Stress Tests Therapeutic Toxic effect Toxicology Toxin Training Water Work base cell injury cheminformatics design drug discovery environmental chemical exposed human population extracellular high throughput screening innovation insight instrument mitochondrial dysfunction mitochondrial permeability transition pore novel small molecule small molecule libraries toxicant

项目摘要

DESCRIPTION (provided by applicant): Many environmental, pharmaceutical, and industrial compounds negatively affect human health by deleterious effects on mitochondrial function. Mitochondrial toxicity is defined as a decrease in the function and /or number of mitochondria, leading to decreased respiration and energy production. Severe loss of mitochondrial function can result in damage to vital organs due to cellular injury, apoptosis, or necrosis. Currently there is no database of chemically related compounds that cause mitochondrial dysfunction, and there are no reliable methods for predicting mitochondrial toxicity. To develop a database of mitochondrial toxicants, a novel respirometric assay of 1760 diverse compounds was conducted using primary cultures of rabbit renal proximal tubule cells (RPTC) and the Seahorse Biosciences Extracellular Flux (XF) analyzer. The results of this screen identified 22 compounds that diminish uncoupled mitochondrial respiration, a stress test for mitochondrial function. From these 22 compounds, five chemical clusters were identified based on molecular similarity between three or more compounds. One of these clusters has been aligned to identify a preliminary group of chemical structures related to mitochondrial toxicity, defined as a toxicophore. We hypothesize that discrete toxicophores defined by distinct chemical entities can identify previously unknown and future mitochondrial toxicants. We will examine this hypothesis in three Specific Aims. Specific Aim 1 is to elucidate the toxicophores for the five identified clusters of mitochondrial toxicants using cheminformatic analysis. Specific Aim 2 is to identify mitochondrial toxicants related to the elucidated toxicophores through an in silico screen of a diverse library of 50,000 small molecules. The resulting compounds will be examined for their effects on mitochondrial respiration using RPTC and a 96-well XF instrument. Confirmed mitochondrial toxicants will be included into the cheminformatic models and used to refine the toxicophores. Specific Aim 3 is to analyze the mechanism of mitochondrial damage for the identified toxicophores by examining the effects of representative compounds on uncoupling oxidative phosphorylation, disruption of the electron transport chain, and/or altering the mitochondrial permeability transition pore (MPT). The long-term goal of this research is to design toxicophores that can reliably predict mitochondrial toxicity for both chemical discovery and risk assessment. The training that I will acquire by completing the outlined proposal will enhance my research expertise, and will prepare me to apply innovative concepts to the field of toxicology. PUBLIC HEALTH RELEVANCE: We hypothesize that discrete toxicophores defined by distinct chemical entities can identify previously unknown and future mitochondrial toxicants. A high-throughput respirometric screen of a diverse chemical library in primary cultures of rabbit renal proximal tubule cells identified five clusters of molecularly similar mitochondrial toxicants. The identified clusters will be used to define toxicophores that can reliably predict mitochondrial toxicity for chemical design, drug discovery, and human health risk assessment.

描述（由申请人提供）：许多环境、药物和工业化合物通过对线粒体功能产生有害影响而对人类健康产生负面影响。线粒体毒性定义为线粒体功能和/或数量减少，导致呼吸和能量产生减少。线粒体功能的严重丧失可能会因细胞损伤、细胞凋亡或坏死而导致重要器官受损。目前还没有导致线粒体功能障碍的化学相关化合物的数据库，也没有可靠的方法来预测线粒体毒性。为了开发线粒体毒物数据库，使用兔肾近端小管细胞 (RPTC) 的原代培养物和 Seahorse Biosciences 细胞外通量 (XF) 分析仪对 1760 种不同化合物进行了新型呼吸测定。该筛选的结果确定了 22 种可减少解偶联线粒体呼吸（线粒体功能的压力测试）的化合物。根据三种或更多化合物之间的分子相似性，从这 22 种化合物中鉴定出了 5 种化学簇。其中一个簇已被对齐，以确定一组与线粒体毒性相关的初步化学结构，定义为毒载体。我们假设由不同化学实体定义的离散毒物团可以识别以前未知和未来的线粒体毒物。我们将在三个具体目标中检验这一假设。具体目标 1 是使用化学信息学分析阐明五种已识别的线粒体毒物簇的毒载体。具体目标 2 是通过对 50,000 个小分子的多样化库进行计算机筛选，识别与已阐明的毒物相关的线粒体毒物。将使用 RPTC 和 96 孔 XF 仪器检查所得化合物对线粒体呼吸的影响。已确认的线粒体毒物将被纳入化学信息学模型并用于细化毒物团。具体目标 3 是通过检查代表性化合物对解偶联氧化磷酸化、破坏电子传递链和/或改变线粒体通透性转换孔 (MPT) 的影响，分析已识别毒物的线粒体损伤机制。这项研究的长期目标是设计能够可靠预测线粒体毒性的毒物载体，以用于化学发现和风险评估。我通过完成概述的提案将获得的培训将增强我的研究专业知识，并使我为将创新概念应用于毒理学领域做好准备。公共卫生相关性：我们假设由不同化学实体定义的离散毒物团可以识别以前未知和未来的线粒体毒物。对兔肾近端小管细胞原代培养物中的多种化学文库进行高通量呼吸测量筛选，鉴定出五簇分子相似的线粒体毒物。确定的簇将用于定义毒物团，可以可靠地预测化学设计、药物发现和人类健康风险评估的线粒体毒性。