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：
8092881
负责人：
SAMSON JAMESDANIEL
金额：
$ 15.19万
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 2013-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8092881
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 Neurodegenerative Disorders 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翻译后修饰，可能导致灾难性后果，具体取决于硝酸蛋白的功能活性。我们报道了顺铂诱导的两个显着耳蜗硝化的证据。这些蛋白质的功能作用是用抗硝基酪氨酸和耳托药物阻断硝化的能力来提出这些蛋白质的功能和神经元染色。这项研究的长期目的是了解蛋白质硝化在耳毒性中的作用，并发现更好的介入药物来预防耳毒性。该提案的具体目的是1）确定人工耳蜗蛋白的硝化及其与听力损失的相关性2）建立身份，以建立身份，主要硝化的耳蜗蛋白（S）和3）使用抑制剂来操纵硝化蛋白，并与抑制剂相关，并与预防性蛋白毒素相关。听觉脑干反应和失真产物耳声发射将用于评估听力功能，而授课图将用于评估形态学损害。硝化蛋白在耳蜗中的定量和定位将分别通过免疫印迹和免疫细胞化学，并用抗硝基酪氨酸的抗体进行免疫印迹和免疫细胞化学。硝化蛋白将通过凝胶电泳鉴定，然后通过MALDI-TOF质谱法鉴定。将使用抗体微阵列研究与硝化相关的蛋白质组学反应，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作用在顺铂介导的核能中的作用，并将促进鉴定有效的治疗干预靶标。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后翻译后修饰，可以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听力功能的相关丧失之间的相关性。通过顺铂治疗硝化的硝化蛋白质的鉴定，并分析随附的人工耳蜗蛋白质组学反应，并有或不抑制硝化作用，将为顺铂毒素毒性的分子机制提供更明显的清晰度。这些反过来将促进高效治疗靶标的鉴定和开发，以防止顺铂介导的耳毒性。