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.

摘要 在过去的100年里，硫芥子气（SM）被反复用作化学武器，最近一次是 2016年在叙利亚。SM引起的急性眼损伤包括严重的眼痛、角膜异常、眼恐怖症， 水肿和失明。晚期慢性毒性还包括芥子气性角膜病变、血管迂曲、角膜缘炎、角膜炎和角膜炎。 缺血和视网膜损伤。目前，还没有描述早期和晚期SM的全面动物研究 对视网膜功能存在毒性。最近的一项人类临床研究报告了视网膜病变的显著减少， 伊朗退伍军人暴露于SM的功能，表现出视网膜内层，双极细胞， Müller胶质细胞。我们观察了暴露于SM的伊朗退伍军人的兔视网膜的体内损伤， 通过1U 01 EY 031650赠款进行角膜抵消研究。试验兔体内数据如下： 用人Müller神经胶质细胞（MIO-M1）的体外对照模型验证， 涉及炎症环境、氧化应激和通过caspase-1/NLRP 3焦亡的细胞死亡。总的来说， 从兔体内和Müller胶质细胞体外研究中收集的初步数据表明，Müller胶质细胞 暴露于芥子气引起严重的时间和剂量依赖性胶质细胞增生的兔眼暴露于SM，和 体外培养Müller胶质细胞。该领域的一个主要知识缺口是缺乏对SM机制的理解 神经视网膜的毒性。人临床、兔体内和Müller细胞体外研究导致了一个中心 假设SM暴露于眼睛引起视网膜双相损伤， 通过神经胶质细胞过度活化的免疫反应和通过神经胶质细胞过度活化的长期神经变性功能性疾病 有缺陷的线粒体自噬这个提议测试了两个新的假设：（a）SM破坏了空间神经框架， 在小型猪模型中体内引起视网膜功能障碍，和（B）SM通过激活引起先天性免疫应答 视网膜小胶质细胞和Müller胶质细胞中NLRP 3炎性小体和线粒体自噬缺陷以及视网膜导线 小型猪眼神经变性。两个具体的目的测试这些假设使用体内哥廷根小型猪SM 与MRI Global（堪萨斯城，MO）和原代猪小胶质细胞（pMicroglia）培养物合作的毒性模型 模型（在我们的实验室中公开/建立）和市售的人Müller神经胶质细胞（MIO-M1细胞）。 目的-1将在体内的临床特点，结构和功能的变化，在视网膜的小型猪模型， 芥子气毒性使用最先进的多模态2D/3D临床成像工具和功能测定。目标- 2将确定参与视网膜变性机制的分子靶点，这些机制是由先天性视网膜色素变性激活引起的。 免疫应答和有缺陷的线粒体自噬，使用细胞和生物化学方法， 视网膜小胶质细胞/Müller胶质细胞和体内哥廷根小型猪视网膜。这个项目的顺利完成 将提供有关视网膜神经结构和神经元功能，视网膜 神经退行性疾病、先天免疫反应和SM暴露后的线粒体自噬缺陷。我们的团队已被 在眼科领域拥有丰富的经验、技能、联合出版物和赠款。