Mechanisms underlying mustard gas-induced conjunctival injury and use of lipid mediators as medical countermeasures

芥子气引起的结膜损伤的机制以及脂质介质作为医疗对策的使用

• 批准号: 10882060
• 负责人: Darlene A Dartt
• 金额: $ 58.33万
• 依托单位: SCHEPENS EYE RESEARCH INSTITUTE
• 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/10882060
• 关键词: Acceleration; Address; Architecture; Biological Assay; CASP1 gene; Cell Culture Techniques; Cell Density; Cell physiology; Cell surface; Chronic; Clinical Management; Clinical assessments; Coculture Techniques; Conjunctival Epithelium; Conjunctivitis; Cornea; Dendritic Cells; Enzyme-Linked Immunosorbent Assay; Epithelial Cells; Exposure to; Eye; Eye Injuries; Flow Cytometry; Fluorescein; Goblet Cells; Histologic; Histopathology; Homeostasis; Human; Immune; In Vitro; Inflammasome; Inflammation; Inflammatory; Inflammatory Response; Injections; Injury; Innate Immune Response; Interleukin-1 beta; Interleukin-6; Lipids; Lymphocytic Infiltrate; MUC5AC gene; Maintenance; Measurement; Measures; Mechlorethamine; Mediating; Mediator; Medical; Methods; Molecular; Mucins; Mucous Membrane; Mus; Mustard Gas; Neuropeptides; Nociceptors; Pathology; Photophobia; Property; Proteins; Regulation; Regulatory T-Lymphocyte; Reporting; Resolution; Reverse Transcriptase Polymerase Chain Reaction; Secretory Cell; Security; Series; Shapes; Signal Transduction; Signs and Symptoms; Site; Small Interfering RNA; Source; Stains; Stimulus; Structure of trigeminal ganglion; Symptoms; Therapeutic; Thrombospondin 1; Tissues; Topical application; Toxic Actions; Toxic effect; Transcript; Transforming Growth Factor beta; Treatment Efficacy; Vesicants; Western Blotting; acute symptom; adaptive immune response; cell injury; conjunctiva; eye dryness; immunoregulation; in vivo; inhibitor; insight; lipid mediator; lipoxin A4; medical countermeasure; mouse model; nerve supply; novel; novel strategies; ocular pain; ocular surface; pharmacologic; prevent; response; restoration; retinal toxicity; slit lamp imaging; therapeutic target; vascular abnormality

PROJECT SUMMARY Mustard gas (MG, most commonly sulfur and nitrogen mustard) is a highly toxic vesicant that causes ocular injuries in over 90% of exposed victims with symptoms lasting from minutes to years. These symptoms include conjunctivitis in addition to eye pain, photophobia and corneal epitheliopathy. Moderate to severe exposure to MG results in chronic or latent (delayed-onset) forms of symptoms that include changes in the conjunctiva such as vascular abnormalities and histopathology including reduced goblet cell (GC) density and lymphocytic infiltration. However, mechanisms of toxicity and therapeutic targets in the conjunctiva remain largely unknown. We reported that in addition to protective soluble mucin, MUC5AC,GCs are a source of pro-inflammatory IL-1β as well as immunomodulatory TGFβ and therefore play a role in shaping both local innate and adaptive immune responses. The contribution of GC secretory products to ocular mucosal homeostasis is further underscored by the observed chronic inflammation and related epitheliopathy associated with loss of GCs or GC-derived regulatory factors. Specialized pro-resolving mediators (SPM) like D-series resolvins RvD1 are endogenous lipid derivatives produced during inflammation are known to accelerate its resolution and restoration of tissue architecture. We reported that RvD1 can inhibit secretion of GC-derived IL-1β. In this proposal we will identify mechanisms underlying MG toxicity in the conjunctiva and its contribution to chronic pathology. We will determine if SPMs restore homeostasis in the conjunctiva to prevent chronic damage caused by MG and serve as effective medical counter measure (MCM) for ocular toxicity of MG. We hypothesize that SPMs promote resolution of MG-induced ocular damage by modulating secretory and homeostatic function of conjunctival GCs. To address this hypothesis, the following three specific aims are proposed. Aim 1: Identify nitrogen mustard (NM)-induced alteration in conjunctival epithelial GC function and the potential of SPMs to resolve NM toxicity and restore GC function in vitro; Aim 2: Determine if NM exposure disrupts immune homeostatic function of GCs in vitro and if SPMs can restore it; and Aim3: Determine therapeutic efficacy of RvD1 as a MCM, in vivo, using a mouse model of NM toxicity. Conjunctival and corneal damage as well as ocular surface innervation will be evaluated. Our anticipated results from these studies can reveal the potential of conjunctival GCs as a MG injury site and therefore a therapeutic target and provide critical insights into developing novel strategies to promote mucosal and immune homeostasis in clinical management of MG-induced ocular toxicity.

项目摘要 芥子气（MG，最常见的是硫和氮芥子气）是一种高毒性的起泡剂， 超过90%的暴露受害者受伤，症状持续数分钟至数年。这些症状包括 结膜炎以及眼痛、畏光和角膜上皮病。中度至重度暴露于 MG导致慢性或潜伏（延迟发作）形式的症状，包括结膜变化， 血管异常和组织病理学，包括杯状细胞（GC）密度降低和淋巴细胞 浸润然而，结膜中的毒性机制和治疗靶点在很大程度上仍然未知。 我们报道了除了保护性的可溶性粘蛋白MUC 5AC，GC是促炎性IL-1β的来源， 以及免疫调节性TGFβ，因此在形成局部先天性和适应性 免疫反应。GC分泌产物对眼粘膜稳态的作用进一步被证实。 通过观察到的与GC损失相关的慢性炎症和相关上皮病变强调，或 GC衍生的调节因子。专门的促消退介质（SPM），如D系列消退素RvD 1， 已知在炎症期间产生的内源性脂质衍生物加速其消退， 组织结构的恢复。我们报道了RvD 1可以抑制GC源性IL-1β的分泌。在这 我们将确定结膜中MG毒性的潜在机制及其对慢性炎症的贡献。 病理我们将确定SPM是否能恢复结膜的稳态，以防止慢性损伤 为MG眼毒性的有效医学对策提供了新的思路。我们 假设SPM通过调节分泌和分泌来促进MG诱导眼损伤的消退， 结膜GC的稳态功能。为了解决这一假设，以下三个具体目标是： 提出了目的1：鉴定氮芥（NM）诱导的结膜上皮GC功能的改变， SPM在体外消除NM毒性和恢复GC功能的潜力;目的2：确定NM暴露是否 在体外破坏GC的免疫稳态功能，以及SPM是否可以恢复它;以及Aim 3：确定 使用NM毒性的小鼠模型，在体内观察RvD 1作为MCM的治疗功效。结膜和角膜 将评价损伤以及眼表面神经支配。这些研究的预期结果可以 揭示了结膜GC作为MG损伤部位的潜力，因此是治疗靶点，并提供了 在临床中开发新策略以促进粘膜和免疫稳态的重要见解 MG诱导的眼毒性的管理。