Role of protein nitration in cisplatin mediated ototoxicity

蛋白质硝化在顺铂介导的耳毒性中的作用

基本信息

批准号：
7714673
负责人：
SAMSON JAMESDANIEL
金额：
$ 15.85万
依托单位：
STATE UNIVERSITY OF NEW YORK AT BUFFALO
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-07-01 至 2012-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7714673
关键词：
3-nitrotyrosine Address Adverse effects Antibodies Antineoplastic Agents Apical Apoptosis Attenuated Auditory Brainstem Responses Biological Biological Process Cardiovascular Diseases Cell Death Cisplatin Cochlea Computer software DNA Adduction Development Dose Dose-Limiting Electrophoresis Evoked Potentials, Auditory, Brain Stem Generations Goals Hair Cells Hearing Immunoblotting Inflammatory Labyrinth Lead Location MALDI-TOF Mass Spectrometry Measures Mediating Modification Molecular Nerve Degeneration Neurons Nitrates Outcome Study Outer Hair Cells Pathology Pathway Analysis Pathway interactions Pharmaceutical Preparations Phosphorylation Physiological Post-Translational Protein Processing Prevention Proteins Proteomics Reaction Time Reactive Oxygen Species Receptor Cell Reporting Research Role Signal Pathway Signaling Protein Staining method Stains Supporting Cell Testing Therapeutic Intervention Time Toxic effect Work base cell type clinically significant design gel electrophoresis hearing impairment immunocytochemistry inhibitor/antagonist nephrotoxicity nitration novel otoacoustic emission ototoxicity prevent protein function public health relevance response therapeutic development therapeutic target

项目摘要

DESCRIPTION (provided by applicant): Nitration of cochlear proteins indicates oxidative posttranslational modification that can lead to catastrophic consequences, depending upon the functional activity of the nitrated proteins. We have reported evidence for cisplatin-induced nitration of two prominent cochlear bands. A functional role for these proteins is suggested by sterocilia and neuronal staining with anti-nitrotyrosine and by the ability of an otoprotective drug to block nitration. The long term aim of this research is to understand the role of protein nitration in ototoxicity and discover better interventional drugs that prevent ototoxicity. The specific aims of this proposal are 1) to determine cisplatin-induced nitration of cochlear proteins and its correlation with hearing loss 2) to establish the identity the major nitrated cochlear protein(s) and 3) to manipulate the nitrated proteins using inhibitor and correlate with prevention of ototoxicity. Auditory brain stem response and distortion product otoacoustic emissions will be used to evaluate the hearing function, while cochleograms will be used to assess morphological damage. Quantification and localization of nitrated proteins in the cochlea will be done by immunoblotting and immunocytochemistry, respectively, with antibodies against nitrotyrosine. Nitrated proteins will be identified by gel electrophoresis followed by MALDI-TOF mass spectrometry. Proteomic responses associated with nitration will be investigated using antibody microarray and analyzed with pathway analysis software. The outcome of this study will reveal the role of cochlear protein nitration in cisplatin mediated ototxicity and will facilitate the identification of effective targets for therapeutic intervention. PUBLIC HEALTH RELEVANCE: Ototoxicity is one among the major dose-limiting factors of the widely used antineoplastic drug cisplatin. Nitration of proteins is considered as an important oxidative posttranslational modification that can lead to serious alterations in protein function. This study will reveal the correlation between cisplatin induced nitration of cochlear proteins and associated loss of hearing function. Identification of the cochlear proteins nitrated by cisplatin treatment and analysis of accompanying cochlear proteomic responses, with and without inhibition of nitration, will add more clarity to the molecular mechanism underlying cisplatin ototoxicity. These in turn will facililitate the identification and development of highly effective therapeutic targets to prevent cisplatin mediated ototoxicity.

描述（由申请人提供）：耳蜗蛋白的硝化表明氧化翻译后修饰可能导致灾难性后果，具体取决于硝化蛋白的功能活性。我们报告了顺铂诱导两个显着耳蜗带硝化的证据。通过抗硝基酪氨酸的纤毛和神经元染色以及耳保护药物阻断硝化的能力表明了这些蛋白质的功能作用。这项研究的长期目标是了解蛋白质硝化在耳毒性中的作用，并发现更好的预防耳毒性的介入药物。该提案的具体目标是 1) 确定顺铂诱导的耳蜗蛋白硝化及其与听力损失的相关性 2) 确定主要硝化耳蜗蛋白的身份和 3) 使用抑制剂和关联来操纵硝化蛋白具有预防耳毒性的作用。听觉脑干反应和畸变产物耳声发射将用于评估听力功能，而耳蜗图将用于评估形态损伤。耳蜗中硝化蛋白的定量和定位将分别通过免疫印迹和免疫细胞化学使用硝基酪氨酸抗体进行。硝化蛋白质将通过凝胶电泳和 MALDI-TOF 质谱进行鉴定。将使用抗体微阵列研究与硝化相关的蛋白质组反应，并使用途径分析软件进行分析。这项研究的结果将揭示耳蜗蛋白硝化在顺铂介导的耳毒性中的作用，并将有助于确定治疗干预的有效靶点。公共卫生相关性：耳毒性是广泛使用的抗肿瘤药物顺铂的主要剂量限制因素之一。蛋白质的硝化被认为是一种重要的氧化翻译后修饰，可导致蛋白质功能的严重改变。这项研究将揭示顺铂诱导的耳蜗蛋白硝化与相关听力功能丧失之间的相关性。顺铂治疗硝化的耳蜗蛋白的鉴定以及伴随硝化抑制和不抑制硝化的伴随耳蜗蛋白质组反应的分析，将使顺铂耳毒性的分子机制更加清晰。这些反过来将有助于高效治疗靶点的识别和开发，以预防顺铂介导的耳毒性。