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，硫磺芥子气(SM)被反复用作化学武器，最近的一次是 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、畏光、 化脓症和失明。晚期慢性毒性还包括芥子气角膜病变、血管扭曲、角膜缘 缺血和视网膜损伤。目前，还没有全面的动物研究来描述早期和晚期SM 对视网膜功能有毒性。最近的一项人类临床研究报告称，视网膜显著减少 暴露于SM的伊朗退伍军人的功能，表现为视网膜内层、双极细胞和 Müler神经胶质细胞。我们在体内观察了暴露于SM的伊朗退伍军人对兔视网膜的损伤 进行我们的角膜对抗研究，由1U01EY031650资助。实验兔的体内数据是 用人Müler胶质细胞体外对照模型(MIO-M1)验证了SM对视网膜的毒性 涉及炎症环境、氧化应激，以及通过caspase-1/NLRP3上睑下垂导致的细胞死亡。总而言之， 从兔体内和体外Müler胶质细胞收集的初步数据表明，Müler神经胶质细胞 暴露于芥子气可引起暴露于SM的兔眼严重的时间和剂量依赖的胶质细胞增生症，以及 Müler神经胶质细胞体外培养。该领域的一个主要知识空白是缺乏对SM的理解机制 对神经视网膜的毒性。人的临床、兔体内和Müler细胞的体外研究导致了中枢 假设SM暴露于眼睛会对视网膜造成双相损伤，从而启动短期的先天性 神经胶质细胞过度激活和长期神经退行性功能性疾病引起的免疫反应 有缺陷的有丝分裂。这一提议检验了两个新的假设：(A)SM扰乱了空间神经框架和 在小型猪模型中引起体内视网膜功能障碍和(B)SM通过激活引起先天免疫反应 NLRP3炎症体和视网膜小胶质细胞和Müler胶质细胞吞噬缺陷 小型猪眼的神经变性。两个特定的目标使用活体Göttingen微型猪SM来验证这些假说 与密苏里州堪萨斯城MRI Global和原代猪小胶质细胞培养合作的毒性模型 模型(在我们实验室发表/建立)和商业上可用的人类Müler神经胶质细胞(MIO-M1细胞)。 AIM-1将在体内表征小型猪视网膜的临床、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分析的芥子气毒性。目标-- 2将确定与先天激活启动的视网膜退行性变机制有关的分子靶点 使用细胞和生化方法的免疫反应和有丝分裂缺陷 视网膜小胶质细胞/Müler胶质细胞和活体Göttingen小型猪视网膜。这项工程的圆满完成 将提供有关视网膜神经结构和神经功能的必要知识 SM暴露后的神经退行性、先天免疫反应和有丝分裂缺陷。我们队有 在眼科对抗领域有丰富的经验、技能、联合出版物和赠款。