Targeting nitrative stress for treatment of cisplatin ototoxicity

靶向硝化应激治疗顺铂耳毒性

• 批准号: 10587579
• 负责人: SAMSON JAMESDANIEL
• 金额: $ 36.87万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-12-01 至 2027-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10587579
• 关键词: Acquired Deafness; Adaptor Signaling Protein; Adverse effects; Anti-Inflammatory Agents; Antineoplastic Agents; Apoptosis; Apoptotic; Attenuated; Auditory; Binding; Biological; CBA/J Mouse; CRISPR/Cas technology; Cancer Patient; Cell Death; Cell Death Inhibition; Cell Survival; Cisplatin; Clinical; Cochlea; Data; Development; Dose Limiting; Down-Regulation; Drug Targeting; Epithelial Cells; Eye; Foundations; Genes; Genetic Transcription; Goals; Hair Cells; Hearing Protection; Induction of Apoptosis; Inflammatory; Intervention; Kidney; Knock-out; Knowledge; LIM Domain; Link; Mass Spectrum Analysis; Mediating; Messenger RNA; Modeling; Modification; Molecular; Mus; Nitrates; Oral; Organ; Organ of Corti; Outcome; Oxidative Stress; Oxidative Stress Induction; Pancreas; Pathway interactions; Patients; Peroxonitrite; Phosphorylation; Play; Predisposition; Protein Inhibition; Proteins; Proteomics; Reporting; Repression; Research; Role; SCID Mice; STAT protein; STAT1 gene; STAT3 gene; Sensory; Signal Pathway; Signal Transduction; Signaling Protein; Site-Directed Mutagenesis; Stress; Testing; Therapeutic; Treatment Efficacy; Treatment Protocols; Ubiquitination; analog; anticancer activity; catalyst; cisplatin induced hearing loss; clinically relevant; cytotoxicity; effective intervention; hearing impairment; innovation; lactacystin; mutant; new therapeutic target; nitration; novel; novel therapeutic intervention; otoprotectant; ototoxicity; overexpression; permanent hearing loss; pharmacologic; prevent; promoter; protein protein interaction; response; side effect; therapeutically effective; translational applications

Abstract A critical gap exists in understanding how nitrative stress, which has been effectively targeted to inhibit cell death in other models, alters cochlear protein signaling to induce apoptosis in cisplatin-induced ototoxicity. Continued existence of this gap represents an important problem for the 40%-80% of cisplatin-treated cancer patients who suffer with significant and in some cases permanent hearing loss as a result of cisplatin use. Until the underlying nitrative stress mechanism is delineated the promise of this new interventional target for mitigating a dose-limiting side-effect of cisplatin likely will remain unrealized. The long-term goal is to better understand the functional as well as mechanistic role of cochlear nitrative stress in acquired hearing loss. The objective is to delineate signaling pathways by which cisplatin-induced nitrative stress, particularly nitration of cochlear LMO4, facilitates ototoxicity, because cisplatin treatment nitrates and downregulates LMO4 protein. LMO4 is a transcriptional regulator that controls pathways regulating cell survival and cell death. The central hypothesis is that cisplatin-induced nitrative stress downregulates cochlear LMO4 and compromises STAT3- mediated anti-apoptotic signaling to facilitate ototoxicity. Understanding the mechanisms whereby nitrated cochlear LMO4 promotes cisplatin-induced ototoxicity is likely to contribute to the development of strategies to prevent this debilitating adverse effect. Guided by strong preliminary data this study will pursue three specific aims: (1) establish the causal link between cisplatin-induced LMO4 nitration and ototoxicity; (2) determine the effects of cisplatin-induced LMO4 nitration on JAK/STAT signaling; and (3) determine the otoprotective efficacy of pharmacological inhibition of nitration. In Aim 1, cisplatin-induced apoptosis will be analyzed after blocking nitration of LMO4 by site-directed mutagenesis and inhibiting proteasomal degradation of nitrated-LMO4 by lactacystin. The link between LMO4 protein levels and cisplatin-induced ototoxicity will be ascertained by testing cochlear apoptosis/hearing loss in LMO4 knockout and overexpressing mice. In Aim 2, cisplatin- induced changes in protein-protein interactions of cochlear LMO4 will be analyzed using a mass spectrometry- based proteomics approach while JAK/STAT related apoptotic and inflammatory signaling will be analyzed using targeted gene arrays. In Aim 3, the otoprotective efficacy of SRI110, a peroxynitrite decomposition catalyst, will be assessed using CBA/J mice; potential interference of SRI110 with anti-cancer activity of cisplatin will be analyzed using SCID mice. This innovative research departs from the status quo by shifting the focus from oxidative stress to the pivotal role of nitrative stress in cisplatin ototoxicity. Significantly, outcomes are expected to vertically advance understanding of how nitrative stress regulates cochlear apoptosis in cisplatin-induced ototoxicity. Findings will have important translational applications in mitigating cisplatin- induced hearing loss and preventing other otopathologies where nitrative stress plays a crucial role.

抽象的 在理解硝化胁迫如何有效地抑制硝酸压力方面存在关键差距 在其他模型中，细胞死亡会改变人工耳蜗信号传导，以诱导顺铂诱导的耳毒性凋亡。 对于40％-80％的顺铂治疗癌症，持续存在该差距是一个重要的问题 遭受重大且在某些情况下因使用顺铂而永久性听力损失的患者。直到 划定了潜在的硝化应力机制 减轻顺铂的剂量限速副作用可能会保持未实现。长期目标是改善 了解耳蜗硝化应力在获得的听力损失中的功能和机械作用。这 目的是描述顺铂诱导的硝化应激，尤其是硝化的信号通路 人工耳蜗LMO4，促进耳毒性，因为顺铂治疗硝酸盐并下调LMO4蛋白。 LMO4是一种转录调节因子，可控制调节细胞存活和细胞死亡的途径。中央 假设是顺铂诱导的硝化应力下调了人耳蜗LMO4，并损害了STAT3- 介导的抗凋亡信号传导以促进耳毒性。了解硝化的机制 耳蜗LMO4促进顺铂诱导的耳毒性可能有助于发展策略 防止这种使人衰弱的不良影响。在强大的初步数据的指导下，这项研究将追求三个特定的特定 目的：（1）在顺铂诱导的LMO4硝化和耳毒性之间建立因果关系； （2）确定 顺铂诱导的LMO4硝化对JAK/STAT信号的影响； （3）确定眼球保护功效 药理学抑制硝化作用。在AIM 1中，将在阻塞后分析顺铂诱导的凋亡 通过位置定向诱变对LMO4的硝化和抑制硝酸LMO4蛋白酶体降解的硝化作用。 乳酸。 LMO4蛋白水平与顺铂诱导的耳毒性之间的联系将通过 测试LMO4敲除和过表达小鼠的耳蜗凋亡/听力损失。在AIM 2中，顺铂 - 将使用质谱法分析耳蜗LMO4蛋白质蛋白质相互作用的诱导变化 基于蛋白质组学方法，将分析JAK/STAT相关的凋亡和炎症信号传导 使用靶向基因阵列。在AIM 3中，SRI110（过氧亚硝酸盐分解 催化剂将使用CBA/J小鼠进行评估； SRI110对抗癌活性的潜在干扰 将使用SCID小鼠分析顺铂。这项创新的研究通过改变了现状 从氧化应激到硝化应激在顺铂耳毒性中的关键作用。重要的是，结果 期望垂直提高硝化应力如何调节人工耳蜗的理解 顺铂诱导的耳毒性。调查结果将在减轻顺铂 - 诱导的听力损失并防止硝化应力起着至关重要的作用的其他耳病病理学。