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：
10559604
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/10559604
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 Cytoplasm Cytoskeleton Data Dependence Dialysis procedure Disease Dose Limiting Environment Environmental Pollutants Environmental and Occupational Exposure Exhibits Experimental Models Exposure to Fumarates Heat-Shock Proteins 90 Heavy Metals High Pressure Liquid Chromatography Human Incubated Injury Injury to Kidney Keratin Kidney Kidney Diseases Label Link Lipids Mass Spectrum Analysis Measures Mercuric chloride Methods Mitochondria Modeling Modification Molecular Weight Non-Steroidal Anti-Inflammatory Agents Organ Transplantation Patients Pattern Pharmaceutical Preparations Phenotype Plasma Polymyxin B 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 adduct anti-cancer biomarker validation chemotherapeutic agent cytotoxicity environmental agent exosome exposed human population extracellular halogenation improved in vivo kidney cell lipidomics 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：暴露于广泛的环境污染物，包括卤代溶剂，多环芳烃碳氢化合物和重金属会导致肾脏损伤。急性肾脏损伤（AKI）也是剂量限制的一面几类治疗药物的影响，包括许多抗生素，抗病毒药，抗癌药化学治疗剂和NSAIDS。尽管一些高度敏感的蛋白质生物标志物已经近年来，它们仍然与一定程度的肾脏损害有关。可以表明在任何最小伤害之前或之后都可以检测到暴露却是可取的。此外，识别与行动机制更紧密联系的新标记将增强理解机制并改善治疗疗法。我们识别此类机械标记的方法主要集中于线粒体作为常见，早期和灵敏靶标的近端管细胞的敏感靶标的一系列环境污染物和治疗药物。我们将使用近端管状的原发性培养物人肾脏（HPT细胞）作为实验模型系统。我们的总体假设是将HPT细胞暴露于环境或治疗相关的毒物或治疗浓度药物分别将修饰线粒体和其他细胞成分，并释放选定蛋白质和脂质以及代谢物的改变模式。尽管线粒体功能障碍与各种形式的肾脏损伤和疾病已经建立了良好识别基于机械的生物标志物的概念是新颖的。 HPT细胞将用两个环境处理污染物（三氯乙烯代谢物S-（1,2-二氯烯基）-l-甲基[DCVC]和HGCL2）或三个所有靶向肾脏线粒体的治疗剂（Tenofovir disoproxy，Cisplatin和Polymyxin 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确定了三种特定蛋白，一个线粒体（亚硫酸盐氧化酶），一个细胞骨架（角蛋白）和一个细胞质（HSP90）作为潜在的生物标志物，并采用靶向验证方法。具体目标1将采用针对性的方法来确定从HPT细胞作为生物标志物的释放蛋白的效用。我们将测试线粒体亚硫酸盐氧化酶，细胞骨架角蛋白和细胞质HSP90是否释放暴露于肾毒性的肾小管毒性和近端管状毒性。蛋白质的释放将与肾细胞功能和生存力以及其他成熟的生物标志物的参数。具体目标2将集中在HPT细胞中作为生物标志物中的改性（加合或氧化）蛋白质。特定目标3将重点放在发布 Cardiolipins，其他脂质和中间代谢物作为暴露和近端的敏感指标肾毒性的管状毒性。最后，特定目标4将集中于蛋白质的潜在识别在外交体中，包括在目标1和2中确定的那些，作为敏感标记的另一个潜在来源暴露和伤害。