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

基本信息

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
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/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&apos 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

项目摘要

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、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。目前，尚无全面的动物研究描述早期和晚期 SM。存在对视网膜功能的毒性。最近的一项人体临床研究报告了视网膜功能的显着降低。暴露于 SM 的伊朗退伍军人的功能，表现出视网膜内层、双极细胞和我们观察到伊朗退伍军人在接触 SM 时观察到兔子视网膜的体内损伤。通过 1U01EY031650 试验兔子体内数据资助进行我们的角膜抵消研究。通过人类 Müller 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 焦亡引起的细胞死亡。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，并且体外 Müller 胶质细胞该领域的一个主要知识差距是缺乏对 SM 机制的了解。人类临床、兔体内和 Müller 细胞体外研究导致了中枢神经视网膜毒性。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该提议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疾病使用体内哥廷根小型猪 SM 测试了这些假设。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。 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这一项目的成功完成。将提供有关视网膜神经结构和神经功能、视网膜的必要知识我们的团队在接触 SM 后发现了神经退行性、先天免疫反应和线粒体自噬缺陷。在眼科 CounterACT 领域拥有丰富的经验、技能、联合出版物和资助。