Targeting nitrative stress for treatment of cisplatin ototoxicity

靶向硝化应激治疗顺铂耳毒性

基本信息

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
项目类别：
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

项目摘要

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.

抽象的在理解硝化应激如何有效地抑制硝化方面存在着关键的差距。在其他模型中，细胞死亡会改变耳蜗蛋白信号传导，从而在顺铂诱导的耳毒性中诱导细胞凋亡。这种差距的持续存在对于 40%-80% 的顺铂治疗癌症来说是一个重要问题因使用顺铂而遭受严重听力损失（在某些情况下为永久性听力损失）的患者。直到潜在的硝化应激机制描绘了这一新的干预目标的前景减轻顺铂的剂量限制副作用可能仍未实现。长期目标是为了更好了解耳蜗硝化应激在获得性听力损失中的功能和机制作用。这目的是描绘顺铂诱导的硝化应激，特别是硝化的信号通路耳蜗 LMO4 会促进耳毒性，因为顺铂治疗会产生硝酸盐并下调 LMO4 蛋白。 LMO4 是一种转录调节因子，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激下调耳蜗 LMO4 并损害 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制定策略防止这种使人衰弱的不利影响。在强有力的初步数据的指导下，本研究将追求三个具体目标目的：(1) 建立顺铂诱导的 LMO4 硝化与耳毒性之间的因果关系； (2) 确定顺铂诱导的 LMO4 硝化对 JAK/STAT 信号传导的影响； (3) 确定耳保护功效硝化的药理抑制作用。目标1，阻断后分析顺铂诱导的细胞凋亡通过定点诱变硝化 LMO4 并通过抑制硝化 LMO4 的蛋白酶体降解乳胞素。 LMO4 蛋白水平与顺铂诱导的耳毒性之间的联系将通过以下方式确定测试 LMO4 敲除和过表达小鼠的耳蜗凋亡/听力损失。在目标 2 中，顺铂- 耳蜗 LMO4 的蛋白质-蛋白质相互作用的诱导变化将使用质谱法进行分析- 基于蛋白质组学方法，同时将分析 JAK/STAT 相关的细胞凋亡和炎症信号传导使用靶向基因阵列。在目标 3 中，SRI110（一种过氧亚硝酸盐分解物）的耳保护功效催化剂，将使用 CBA/J 小鼠进行评估； SRI110 抗癌活性的潜在干扰将使用 SCID 小鼠分析顺铂。这项创新研究改变了现状，重点从氧化应激转向硝化应激在顺铂耳毒性中的关键作用。值得注意的是，结果预计将垂直推进对硝化应激如何调节耳蜗细胞凋亡的理解顺铂引起的耳毒性。研究结果将在减轻顺铂- 诱发听力损失并预防硝化应激发挥关键作用的其他耳病。