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.

抽象的在过去的100年中，硫芥末气（SM）已反复用作化学武器 2016年在叙利亚。SM急性眼损伤包括严重的眼部疼痛，角膜异常，恐惧症，化学和失明。晚期慢性毒性还包括芥末气的角膜病，血管曲折，末端缺血和视网膜损害。目前，尚无描述早期和晚期SM的全面动物研究存在对视网膜功能的毒性。最近的人类临床研究报告说，视网膜显着减少在暴露于SM的伊朗退伍军人中的功能，在视网膜，双极细胞和 müller神经胶质细胞。我们观察到在暴露于SM的伊朗退伍军人中看到的对兔视网膜的体内损害通过1U01EY031650赠款进行了我们的角膜抵抗研究。 PILOT RABBIT IN VIVO数据是用人类müller神经胶质细胞（MIO-M1）的受控体外模型验证，视网膜中的毒性涉及炎症环境，氧化应激和通过caspase-1/nlrp3凋亡的细胞死亡。共同体内从兔子和穆勒神经胶质细胞中收集的初步数据表明，穆勒神经胶质细胞暴露于芥末气体导致暴露于SM的兔眼中的严重时间和剂量依赖性神经病， Müller神经胶质细胞体外。该领域的主要知识差距是缺乏SM的理解机制神经视网膜的毒性。人类临床，体内兔和穆勒细胞的体外研究导致了中央 SM暴露于眼睛的假设会对视网膜造成双相损害通过过度激活神经胶质细胞和长期神经退行性功能疾病的免疫反应线粒体缺陷。该建议检验了两个新的假设：（a）SM破坏空间神经框架和在Minipig模型中引起视网膜功能障碍，（B）SM通过激活引起先天免疫响应视网膜小胶质细胞和müller神经胶质细胞中NLRP3炎性体和有缺陷的线粒体，并导致视网膜 Minipig眼中的神经变性。两个特定的目的测试了这些假设，使用体内göttingenminipig sm 毒性模型与MRI Global，堪萨斯城，密苏里州和主要的猪小胶质细胞（PMICROGLIA）培养模型（在我们的实验室中发布/建立）和市售的人类神经胶质细胞（MIO-M1细胞）。 AIM-1将表征体内临床，结构和功能变化的Minipig模型的视网膜使用最先进的多模式2D/3D临床成像工具和功能测定的芥末气体毒性。目的- 2将确定由先天性激活引发的视网膜退化机制所涉及的分子靶标使用细胞和生化方法，使用细胞和生化方法，使用体外模型视网膜小胶质细胞/müller神经胶质细胞和体内Minipig视网膜。该项目的成功完成将提供有关永久性神经元结构和神经元功能的必要知识，永久性 SM暴露后神经退行性，先天免疫调查和线索缺陷。我们的团队有大量经验，技能，共同出版物和赠款在眼部抵抗领域。