Unraveling the corneal and retinal mechanisms of chemical injury

揭示化学损伤的角膜和视网膜机制

基本信息

批准号：
10882069
负责人：
ROYCE MOHAN
金额：
$ 49.82万
依托单位：
UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-01 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10882069
关键词：
Achievement Acute Alkalies Anterior Applications Grants Arginine deiminase Automobile Driving Binding Blindness Calcium Cell Culture Techniques Cell Death Characteristics Chemical Exposure Chemical Injury Chronic Cicatrix Clinic Clinical Cornea Corneal Ulcer Corneal pain Cutaneous DNA Damage Data Disease Dose Drug Combinations Drug Targeting Enzymes Epithelial Cells Event Evolution Exposure to Eye Eye Injuries Eye diseases Fibrosis Funding Future Genetic Gliosis Grant Human In Vitro Infection prevention Inflammation Injury Intermediate Filament Proteins Investigation Knockout Mice Laser injury Link Mechlorethamine Mediating Mediator Medical Modeling Modification Molecular Muller&apos s cell Mus Mustard Gas Ocular Pathology Oryctolagus cuniculus Oxides Pathologic Pathology Pathway interactions Pharmaceutical Preparations Poison Post-Translational Protein Processing Protein Isoforms Proteins Proteomics Publishing Rattus Reporting Research Retina Retinal Degeneration Security Severities Sulfur System TP53 gene Testing Therapeutic Tissues Translating Vesicants Visual impairment analog biological adaptation to stress cellular targeting chemical countermeasure chemical threat combat corneal epithelium crosslink drug discovery experimental study human disease in vivo in vivo Model inhibitor injured lewisite lung injury mass casualty novel pharmacologic programs response symptom management weapons

项目摘要

Abstract: Chemical threat agents can potentially be weaponized to cause mass casualties and such egregious events are of grave concern to homeland security. Sulfur mustard and its analog nitrogen mustard (NM) are vesicants that cause severe acute injury to ocular tissues and ultimately develop fibrotic scarring leading to vision loss. As medical treatments for vesicant injury have yet to be developed for paucity of a complete understanding of their pathophysiological mechanisms, this unmet medical need remains an urgent matter of importance to the Chemical Countermeasures Research Program (CCRP). Under an exploratory R21 grant funded by the CCRP, we have discovered that NM injury to the mouse cornea triggers a rapid and profound induction of retinal gliosis. This retinal response is coordinated in reactive Muller glia with activation of the posttranslational modification (PTM) known as citrullination. This finding extends our prior results showing that retinal hypercitrullination driven by the enzyme peptidyl arginine deiminase (PAD)-4 is a shared pathological feature that is common to a number of different injury and disease paradigms. But critical gaps remain regarding the molecular steps involved in the PAD-hypercitrullination axis. As other published reports have shown that NM crosslinks tumor suppressor p53 and the p53 DNA damage pathway interacts with PAD4, these data provokes an interesting idea whether these two major orchestrators of pathological response are tied to activating hypercitrullination in both the cornea and retina. In this R01 grant proposal, we will investigate the acute and chronic (delayed) injury mechanisms of NM across the cornea and retina in mouse and rat models. Specifically in Aim 1, we will investigate the molecular mechanisms of the p53-PAD axis involving corneal hypercitrullination and identify the relevant PAD using both in vivo and cell culture models to study NM injury. In Aim 2, we will characterize the corneal and retinal citrullinomes of NM injury using proteomics by examining the temporal changes over the early and chronic stages. This study will allow us to identify key PAD substrates of acute and chronic injury states, permitting us to decipher the evolution of key protein players modified by citrullination in perpetuating corneal and retinal pathology. We believe this in-depth investigation into hypercitrullination in the NM injured eye could unravel novel mechanisms and druggable PAD targets that would be critical for drug discovery to combat chemical injury to the human eye. Given that PADs are already being investigated as targets for several common diseases, the likelihood of clinical drugs becoming available for humans in the near future makes this an attractive target class for further mechanistic assessment.

抽象的：化学威胁剂有可能被武器化，造成大规模人员伤亡等令人震惊的事件对国土安全构成严重关切。硫芥及其类似物氮芥 (NM) 是一种起泡剂，可对眼组织造成严重急性损伤，最终形成纤维化疤痕，导致视力丧失。作为治疗疱疹性损伤的药物由于缺乏对其病理生理学的完整了解，尚未得到发展机制，这种未满足的医疗需求仍然是化学品管理局的一个紧迫的重要问题对策研究计划（CCRP）。根据由 R21 资助的探索性拨款 CCRP，我们发现小鼠角膜的 NM 损伤会引发快速而深远的损伤诱导视网膜神经胶质细胞增生。这种视网膜反应在反应性米勒神经胶质细胞中与称为瓜氨酸化的翻译后修饰 (PTM) 的激活。这一发现延伸我们之前的结果表明，肽基精氨酸酶驱动的视网膜过度瓜氨酸化脱亚胺酶 (PAD)-4 是许多不同损伤所共有的共同病理特征和疾病范式。但在涉及的分子步骤方面仍然存在关键差距 PAD-高瓜氨酸轴。正如其他已发表的报告所示，NM 交联肿瘤抑制子 p53 和 p53 DNA 损伤途径与 PAD4 相互作用，这些数据引发了有趣的想法是否这两个主要的病理反应的协调者与激活角膜和视网膜的过度瓜氨酸化。在此 R01 拨款提案中，我们将研究 NM 穿过角膜和视网膜的急性和慢性（迟发性）损伤机制在小鼠和大鼠模型中。具体来说，在目标 1 中，我们将研究以下分子机制：涉及角膜过度瓜氨酸化的 p53-PAD 轴，并使用两者来识别相关的 PAD 研究 NM 损伤的体内和细胞培养模型。在目标 2 中，我们将描述角膜和使用蛋白质组学通过检查 NM 损伤的时间变化来研究 NM 损伤的视网膜瓜氨酸组早期和慢性阶段。这项研究将使我们能够确定急性和慢性疾病的关键 PAD 底物。慢性损伤状态，使我们能够破译关键蛋白质参与者的进化瓜氨酸化使角膜和视网膜病理永久化。我们相信这次深入调查研究 NM 损伤眼中的瓜氨酸过度化可能会揭示新的机制和可药物化的 PAD 这些目标对于发现对抗人眼化学损伤的药物至关重要。给定 PAD 已经被作为几种常见疾病的靶点进行研究，可能性在不久的将来，大量的临床药物将可供人类使用，这使其成为一个有吸引力的目标进一步机械评估的课程。