Mechanistic Understanding of Mustard Gas Toxicity in the Retina using a Minipig Model

使用小型猪模型了解芥子气视网膜毒性的机制

• 批准号: 10882080
• 负责人: Shyam Sunder Chaurasia
• 金额: $ 48.22万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 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/10882080
• 关键词: 3-Dimensional; Acute; Affect; Alkylating Agents; Animal Model; Animals; Architecture; Biochemical; Biological Assay; Blindness; Blood Vessels; CASP1 gene; Cell Culture Techniques; Cell Death; Cell Line; Cells; Chemical Weapons; Chronic; Cities; Clinical; Clinical Research; Collaborations; Cornea; Data; Defect; Dose; Electroretinography; Evaluation; Exhibits; Exposure to; Eye; Eye Injuries; Family suidae; Fluorescein Angiography; Functional disorder; Funding; Fundus; Gases; Gliosis; Grant; Human; Imaging Device; In Vitro; Individual; Inflammasome; Innate Immune Response; Ischemia; Joints; Kansas; Keratopathy; Knowledge; Late Effects; Magnetic Resonance Imaging; Measures; Microglia; Miniature Swine; Modeling; Molecular Target; Muller' s cell; Mustard Gas; Nerve Degeneration; Neural Retina; Neuroglia; Neurons; Ophthalmologist; Organ; Oryctolagus cuniculus; Oxidative Stress; Phase; Photophobia; Process; Publications; Publishing; Reporting; Research Personnel; Retina; Retinal Degeneration; Role; Scientist; Syria; System; Techniques; Testing; Time; Tissues; Toxic effect; Transmission Electron Microscopy; Veterans; War; clinical imaging; experience; human model; human study; in vitro Model; in vivo; inflammatory milieu; insight; limbal; multimodality; neural; novel; ocular pain; retinal damage; skills; tool

ABSTRACT Sulfur mustard gas (SM) has been used repeatedly as a chemical weapon in the last 100 years, most recently in Syria in 2016. Acute ocular injury from SM includes severe ocular pain, corneal abnormalities, photophobia, chemosis, and blindness. Late chronic toxicity also includes mustard gas keratopathy, vascular tortuosity, limbal ischemia, and retinal damage. At present, no comprehensive animal study describing early and late SM toxicity to retinal function exists. A recent human clinical study reported a significant reduction in retinal function in Iranian veterans exposed to SM, exhibiting defects in the inner layers of the retina, bipolar cells, and Müller glial cells. We observed in vivo damage to rabbit retina seen in Iranian veterans exposed to SM while conducting our corneal counteract studies funded through 1U01EY031650 grant. Pilot rabbit in vivo data was verified with a controlled in vitro model of human Müller glial cells (MIO-M1), depictingSM toxicity in retina involves inflammatory milieu, oxidative stress, and cell death via caspase-1/NLRP3 pyroptosis. Collectively, preliminary data collected from rabbit in vivo and Müller glial cell in vitro studies suggested that Müller glial cells exposed to mustard gas caused severe time- and dose-dependent gliosis in rabbit eyes exposed to SM, and Müller glial cell in vitro. A major knowledge gap in the field is the lack of understanding mechanism of SM toxicity in the neural retina. The human clinical, rabbit in vivo and Müller cell in vitro studies led to a central hypothesis that SM exposure to the eye causes biphasic damage to the retina actuating short-term innate immune responses via hyperactivation of glial cells and long-term neurodegenerative functional malady via defective mitophagy. This proposal tests two novel hypotheses: (a) SM disrupts spatial neural framework and causes retinal dysfunction in vivo in a minipig model and (b) SM causes innate immune response via activation of NLRP3 inflammasome and defective mitophagy in retinal microglia and Müller glia cells and leads retinal neurodegeneration in minipig eye. Two specific aims test these hypotheses using in vivo Göttingen minipig SM toxicity model in collaboration with MRI Global, Kansas City, MO and a primary pig microglia (pMicroglia) culture model (published/established in our lab) and commercially available human Müller glial cells (MIO-M1 cells). Aim-1 will characterize in vivo clinical, structural, and functional changes in the retina of a minipig model of mustard gas toxicity using state-of-the-art multi-modal 2D/3D clinical imaging tools and functional assays. Aim- 2 will identify molecular targets involved in retinal degenerative mechanisms initiated by the activation of innate immune responses and defective mitophagy using cellular and biochemical approaches using in vitro models of retinal microglia/Müller glial cells and in vivo Göttingen minipig retina. The successful completion of this project will provide necessary knowledge about retinal neural architecture and neuronal function, retinal neurodegenerative, innate immune responses, and defective mitophagy after SM exposure. Our team has extensive experience, skills, joint publications and grants in the ocular CounterACT field.

抽象的 过去 100 年来，芥子气 (SM) 多次被用作化学武器，最近一次是 2016 年在叙利亚。SM 引起的急性眼损伤包括严重眼痛、角膜异常、畏光、 球结膜水肿和失明。晚期慢性毒性还包括芥子气角膜病变、血管迂曲、角膜缘病变 缺血和视网膜损伤。目前，尚无描述早期和晚期 SM 的综合动物研究 存在对视网膜功能的毒性。最近的一项人类临床研究报告称，视网膜色素变性显着降低。 暴露于 SM 的伊朗退伍军人的功能，表现出视网膜内层、双极细胞和 米勒神经胶质细胞。我们观察到暴露于 SM 的伊朗退伍军人体内兔子视网膜受到损伤 通过 1U01EY031650 拨款资助进行角膜抵消研究。试验兔体内数据为 通过人穆勒神经胶质细胞 (MIO-M1) 的受控体外模型进行验证，描述 SM 在视网膜中的毒性 涉及炎症环境、氧化应激和通过 caspase-1/NLRP3 焦亡导致的细胞死亡。总的来说， 从兔子体内和 Müller 胶质细胞体外研究中收集的初步数据表明，Müller 胶质细胞 暴露于芥子气会导致暴露于 SM 的兔子眼睛发生严重的时间和剂量依赖性神经胶质增生，并且 米勒神经胶质细胞体外。该领域的一个主要知识差距是缺乏对 SM 机制的理解 神经视网膜毒性。人类临床、兔体内和 Müller 细胞体外研究得出了一个中心结论 假设 SM 暴露于眼睛会导致视网膜双相损伤，从而激活短期先天性 通过神经胶质细胞过度激活产生的免疫反应和长期神经退行性功能性疾病 有缺陷的线粒体自噬。该提案测试了两个新的假设：（a）SM 破坏了空间神经框架和 在小型猪模型中引起体内视网膜功能障碍，并且 (b) SM 通过激活引起先天免疫反应 视网膜小胶质细胞和穆勒胶质细胞中 NLRP3 炎性体和线粒体自噬缺陷的影响，并导致视网膜 小型猪眼睛的神经退行性变。两个具体目标使用哥廷根小型猪 SM 体内测试这些假设 与密苏里州堪萨斯城 MRI Global 和原代猪小胶质细胞 (pMicroglia) 培养物合作建立毒性模型 模型（在我们的实验室发布/建立）和市售的人类 Müller 胶质细胞（MIO-M1 细胞）。 Aim-1 将表征小型猪模型视网膜的体内临床、结构和功能变化 使用最先进的多模式 2D/3D 临床成像工具和功能测定来检测芥子气毒性。目的- 2 将确定参与由先天性激活引起的视网膜退行性机制的分子靶点 使用细胞和生化方法使用体外模型进行免疫反应和有缺陷的线粒体自噬 视网膜小胶质细胞/穆勒胶质细胞和体内哥廷根小型猪视网膜。本项目的顺利完成 将提供有关视网膜神经结构和神经元功能、视网膜的必要知识 SM 暴露后的神经退行性、先天免疫反应和线粒体自噬缺陷。我们的团队有 在眼科 CounterACT 领域拥有丰富的经验、技能、联合出版物和资助。