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.

摘要 一个关键的差距存在于理解如何nitrative压力，这已被有效地针对抑制 细胞死亡，改变耳蜗蛋白信号传导以诱导顺铂诱导的耳毒性中的细胞凋亡。 这种差距的持续存在代表了40%-80%顺铂治疗的癌症的重要问题 由于使用顺铂而遭受显著和在某些情况下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种新的介入靶点的前景， 减轻顺铂的剂量限制性副作用可能仍然无法实现。长期目标是更好地 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。的 目的是描述顺铂诱导的硝化应激，特别是 耳蜗LMO 4，促进耳毒性，因为顺铂治疗硝酸盐和下调LMO 4蛋白。 LMO 4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 一种假设是顺铂诱导的硝化应激下调耳蜗LMO 4并损害STAT 3- 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LMO 4促进顺铂诱导的耳毒性可能有助于制定策略， 避免这种负面影响。在强有力的初步数据的指导下，这项研究将追求三个具体的 目的：（1）建立顺铂诱导的LMO 4硝化和耳毒性之间的因果关系;（2）确定 顺铂诱导的LMO 4硝化对JAK/STAT信号传导的影响;以及（3）确定耳保护功效 药理学上抑制硝化作用。在目标1中，将在阻断后分析顺铂诱导的细胞凋亡 通过定点诱变硝化LMO 4和通过抑制硝化-LMO 4的蛋白酶体降解 乳胞素LMO 4蛋白水平与顺铂诱导的耳毒性之间的联系将通过以下方法确定： 在LMO 4敲除和过表达小鼠中测试耳蜗细胞凋亡/听力损失。在目标2中，顺铂- 将使用质谱分析耳蜗LMO 4的蛋白质-蛋白质相互作用的诱导变化- 同时分析JAK/STAT相关的凋亡和炎症信号传导 使用靶向基因阵列。在目标3中，SRI 110（过氧亚硝酸根分解物）的耳保护功效被证明是有效的。 将使用CBA/J小鼠评估SRI 110对催化剂的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 重点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 有望垂直推进对硝化应激如何调节耳蜗细胞凋亡的理解， 顺铂诱导的耳毒性。研究结果将在减轻顺铂方面具有重要的转化应用- 诱发的听力损失和预防其他耳病理学，其中硝化应激起着至关重要的作用。