Mitochondrial and Cellular Biomarkers of Renal Injury from Environmental and Therapeutic Agents

环境和治疗药物引起的肾损伤的线粒体和细胞生物标志物

基本信息

批准号：
10388109
负责人：
LAWRENCE H. LASH
金额：
$ 36.9万
依托单位：
WAYNE STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-04-09 至 2025-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10388109
关键词：
Acute Renal Failure with Renal Papillary Necrosis Antibiotics Antimycin A Antiviral Agents Aromatic Polycyclic Hydrocarbons Benchmarking Biological Markers Biological Models Cardiolipins Cell Survival Cell physiology Cells Chemical Exposure Chemicals Chronic Kidney Failure Cisplatin Cysteine Data Dependence Dialysis procedure Disease Dose-Limiting Environmental Pollution Environmental and Occupational Exposure Exhibits Experimental Models Exposure to Fumarates Heat-Shock Proteins 90 Heavy Metals Human Incubated Injury Injury to Kidney Keratin Kidney Kidney Diseases Label Link Lipids Liquid Chromatography Mass Spectrum Analysis Measures Mercuric chloride Methods Mitochondria Modeling Modification Molecular Weight Non-Steroidal Anti-Inflammatory Agents Organ Transplantation Oxides Patients Pattern Performance Pharmaceutical Preparations Phenotype Plasma Polymyxin B Post-Translational Protein Processing Proteins Proteomics Public Health Rattus Recovery Renal Replacement Therapy Resolution Shotguns Solvents Source Spectrometry, Mass, Electrospray Ionization Supporting Cell Tenofovir Testing Therapeutic Therapeutic Agents Therapeutic Uses Time Toxic effect Trichloroethylene Tubular formation Urine Validation anti-cancer base chemotherapeutic agent cytotoxicity environmental agent exosome exposed human population extracellular improved in vivo kidney cell lipidomics liquid chromatography mass spectrometry mitochondrial dysfunction multiple reaction monitoring nephrotoxicity novel novel marker potential biomarker protein biomarkers renal damage side effect sulfite oxidase tandem mass spectrometry toxicant

项目摘要

Abstract: Exposure to a broad range of environmental contaminants, including halogenated solvents, polycyclic aromatic hydrocarbons, and heavy metals, can cause kidney injury. Acute kidney injury (AKI) is also a dose-limiting side effect of several classes of therapeutic drugs, including many antibiotics, antiviral agents, anticancer chemotherapeutic agents, and NSAIDs. Although some highly sensitive protein biomarkers have been validated in recent years, they are still associated with some degree of renal damage. Markers that can indicate exposure yet be detected prior to any or after only minimal injury are preferable. Additionally, identification of new markers that are more closely linked to mechanism of action will enhance understanding of mechanism and improve therapeutics. Our approach to identification of such mechanistic markers has focused primarily on the mitochondria as common, early, and sensitive targets in proximal tubular cells for an array of environmental contaminants and therapeutic drugs. We will use primary cultures of proximal tubular cells from human kidneys (hPT cells) as the experimental model system. Our overall hypothesis is that exposure of hPT cells to environmentally or therapeutically relevant concentrations of toxicants or therapeutic agents, respectively, will modify mitochondria and other cellular components and result in release of selected proteins and lipids and altered patterns of metabolites. Although association of mitochondrial dysfunction with various forms of kidney injury and disease is well-established, our application of this central, underlying concept to identify mechanistically-based biomarkers is novel. hPT cells will be treated with two environmental contaminants (the trichloroethylene metabolite S-(1,2-dichlorovinyl)-L-cysteine [DCVC] and HgCl2) or three therapeutic agents (tenofovir disoproxil fumarate, cisplatin, and polymyxin B) that all target renal mitochondria. Additionally, antimycin A will be used as a positive control. Preliminary studies helped refine the hypothesis and identified three specific proteins, one mitochondrial (sulfite oxidase), one cytoskeletal (keratins), and one cytoplasmic (HSP90), as potential biomarkers and take a targeted approach for validation. Specific Aim 1 will take a targeted approach to determine the utility of released proteins from hPT cells as biomarkers. We will test whether release of mitochondrial sulfite oxidase, cytoskeletal keratins, and cytoplasmic HSP90 reflect exposure to and proximal tubular toxicity from nephrotoxicants. Release of proteins will be correlated with parameters of renal cell function and viability and other well-established biomarkers. Specific Aim 2 will focus on modified (adducted or oxidized) proteins in hPT cells as biomarkers. Specific Aim 3 will focus on release of cardiolipins, other lipids, and intermediary metabolites as sensitive indicators of exposure to and proximal tubular toxicity from nephrotoxicants. Finally, Specific Aim 4 will focus on the potential identification of proteins in exposomes, including those identified in Aims 1 and 2, as another potential source of sensitive markers of exposure and injury.

抽象的：暴露于多种环境污染物，包括卤化溶剂、多环芳香族化合物碳氢化合物和重金属会导致肾脏损伤。急性肾损伤（AKI）也是剂量限制方几类治疗药物的作用，包括许多抗生素、抗病毒药物、抗癌药物化疗药物和非甾体抗炎药。尽管一些高度敏感的蛋白质生物标志物已被近年来经过验证，它们仍与一定程度的肾损伤有关。可以的标记最好在任何伤害之前或之后表明暴露但已被检测到。此外，鉴定与作用机制更密切相关的新标记将增强理解机制并改进治疗方法。我们识别此类机械标记的方法主要关注线粒体作为近端肾小管细胞中常见、早期和敏感的靶标一系列环境污染物和治疗药物。我们将使用近端肾小管的原代培养物来自人类肾脏的细胞（hPT细胞）作为实验模型系统。我们的总体假设是 hPT 细胞暴露于环境或治疗相关浓度的毒物或治疗剂药物将分别修饰线粒体和其他细胞成分，并导致选定的释放蛋白质和脂质以及代谢物模式的改变。尽管线粒体功能障碍与各种形式的肾损伤和疾病已经得到证实，我们对这一核心的、潜在的应用识别基于机械的生物标志物的概念是新颖的。 hPT 细胞将接受两种环境处理污染物（三氯乙烯代谢物 S-(1,2-二氯乙烯基)-L-半胱氨酸 [DCVC] 和 HgCl2）或三种均针对肾线粒体的治疗药物（富马酸替诺福韦二吡呋酯、顺铂和多粘菌素 B）。另外，抗霉素A将用作阳性对照。初步研究有助于完善假设并鉴定出三种特定蛋白质，一种线粒体（亚硫酸氧化酶）、一种细胞骨架（角蛋白）和一种细胞质（HSP90）作为潜在的生物标志物并采取有针对性的方法进行验证。具体目标 1 将采用有针对性的方法来确定 hPT 细胞释放的蛋白质作为生物标志物的效用。我们将测试线粒体亚硫酸氧化酶、细胞骨架角蛋白和细胞质 HSP90 的释放是否反映暴露于肾毒物和近端肾小管毒性。蛋白质的释放将与肾细胞功能和活力参数以及其他成熟的生物标志物。具体目标2将重点关注 hPT 细胞中修饰（加合或氧化）蛋白质作为生物标志物。具体目标 3 将侧重于发布心磷脂、其他脂质和中间代谢物作为暴露和近端的敏感指标肾毒物引起的肾小管毒性。最后，具体目标 4 将重点关注蛋白质的潜在鉴定在暴露体中，包括目标 1 和 2 中确定的那些，作为敏感标记物的另一个潜在来源暴露和伤害。