The role of galectin-8 in the regulation of corneal infection and inflammation

Galectin-8在调节角膜感染和炎症中的作用

• 批准号: 10186753
• 负责人: Noorjahan Panjwani
• 金额: $ 40.01万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-30 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10186753
• 关键词: Affect; Animals; Bacteria; Bacterial Infections; Binding; Biochemical; Blindness; CASP1 gene; Cells; Cleaved cell; Communicable Diseases; Contact Lenses; Cornea; Corneal Diseases; Data; Development; Disease; Dry Eye Syndromes; Galactose Binding Lectin; Gene Expression Profiling; Generations; Goals; Graft Rejection; Gram-Negative Bacteria; Immune response; Immunologics; In Vitro; Individual; Infection; Inflammasome; Inflammation; Inflammatory; Inflammatory Response; Injections; Innate Immune Response; Innate Immune System; Interleukin-1 beta; Interleukin-18; Invaded; Keratitis; Keratoplasty; Knock-out; Knockout Mice; Leukocyte Elastase; Mediating; Microbe; Modeling; Molecular; Mus; Neutrophil Infiltration; Outcome; Pathogenesis; Pathogenicity; Pathway interactions; Pattern recognition receptor; Pilot Projects; Play; Prevention strategy; Process; Proteome; Pseudomonas; Pseudomonas aeruginosa; Pseudomonas aeruginosa infection; Reaction; Regulation; Resistance; Risk Factors; Role; Serine Protease; Signal Transduction; TLR2 gene; TLR4 gene; TLR5 gene; Testing; United States; Visual impairment; Wild Type Mouse; autoimmune uveitis; base; cell type; chemokine; cytokine; design; expectation; experimental study; immunopathology; knockout animal; macrophage; microbial; mouse model; neutrophil; neutrophil elastase inhibitor; novel; pathogen; transcriptome; transcriptome sequencing

Project Summary The bacterium Pseudomonas aeruginosa (PA) is a leading cause of microbial infection of the cornea. One of the serious consequences of PA corneal infection is blindness resulting from persistent corneal inflammation. Approximately 30,000 cases of microbial keratitis occur annually in the United States. The innate immune system is the first line of defense against pathogens and is initiated by pattern recognition receptors which respond to invading microbes. It is known that IL-1β plays an important role in the induction of immune response in PA keratitis. In the classical immune response to bacterial infection, generation of mature IL-1β is a two-step process. The first step is the induction of pro-IL-1β expression, which is generally achieved by TLR-mediated activation of NF-B pathway that results in the induction of pro-IL-1β. A second signal then triggers the assembly of inflammasomes leading to the cleavage of caspase-1. Active (cleaved) caspase-1 cleaves pro-IL-1β to generate active IL-1β, which is secreted from the cell to mediate downstream inflammatory effects that clear the infection. Recently, we have made an exciting observation that galectin-8 knockout (Gal-8 KO) mice are resistant to PA infection. Additional pilot studies revealed that Gal-8 has the potential to regulate the activation of both TLR and inflammasome pathways. The goal of this project is to characterize the role of Gal-8 in the regulation of PA-induced immunopathology, and to develop effective strategies for prevention and regression of uncontrolled inflammatory response that results in extensive damage to the cornea and visual impairment. In Aim 1, in an effort to understand the mechanisms that render Gal-8 KO mice resistant to PA keratitis, we will test the hypothesis that Gal-8 has the capacity to dampen inflammasome-mediated generation of mature IL-1β to influence innate immune response. Specifically, in this Aim, using Gal-8 KO mice and Gal-8 deficient macrophages and neutrophils, we will characterize the role of Gal-8 in the regulation of the inflammasome pathway in PA keratitis. In Aim 2, we will determine whether Gal-8 influences the outcome of PA keratitis by modulating neutrophil elastase and/or TLR pathway. To better characterize the role of Gal-8 in the pathogenesis of PA keratitis, in Aim 3A, we propose to perform transcriptome and cytokine/chemokine proteome analyses of PA-infected corneas of WT and Gal-8 KO mice, and WT and Gal-8 KO macrophages and neutrophils. In Aim 3B, studies will be performed to determine whether Gal-8 can be targeted to control overactive immune response and corneal inflammation in the mouse model of PA keratitis. We expect that the proposed study will provide ground-breaking data, not only for understanding the molecular mechanism of PA keratitis, but also for the development of novel, galectin-based therapy that can treat blinding immunopathology resulting both from bacterial keratitis, as well as from other ocular disorders, such as corneal graft rejection and dry eye disease.

项目概要 铜绿假单胞菌 (PA) 是角膜微生物感染的主要原因。中的一个 PA角膜感染的严重后果是持续性角膜炎症导致失明。 美国每年大约发生 30,000 例微生物性角膜炎病例。先天免疫系统 是对抗病原体的第一道防线，由模式识别受体启动，该受体对 入侵微生物。众所周知，IL-1β在诱导PA免疫反应中起重要作用 角膜炎。在针对细菌感染的经典免疫反应中，成熟 IL-1β 的产生分两步进行 过程。第一步是诱导pro-IL-1β表达，这通常是通过TLR介导来实现的 NF-κB 通路的激活导致 IL-1β 前体的诱导。然后第二个信号触发组件 炎症小体导致 caspase-1 裂解。活性（裂解的）caspase-1 将 pro-IL-1β 裂解为 产生活性 IL-1β，由细胞分泌，介导下游炎症效应，从而清除 感染。最近，我们做了一个令人兴奋的观察，半乳糖凝集素8敲除（Gal-8 KO）小鼠具有耐药性 PA 感染。其他初步研究表明 Gal-8 有潜力调节这两种物质的激活 TLR 和炎症小体途径。该项目的目标是描述 Gal-8 在调节中的作用 PA 诱导的免疫病理学，并制定有效的策略来预防和消退 不受控制的炎症反应，导致角膜广泛损伤和视力障碍。在 目标 1，为了了解 Gal-8 KO 小鼠对 PA 角膜炎具有抵抗力的机制，我们将测试 假设 Gal-8 有能力抑制炎症小体介导的成熟 IL-1β 的产生 影响先天免疫反应。具体来说，在这个目标中，使用 Gal-8 KO 小鼠和 Gal-8 缺陷小鼠 巨噬细胞和中性粒细胞，我们将描述 Gal-8 在炎症小体调节中的作用 PA 角膜炎的途径。在目标 2 中，我们将通过以下方式确定 Gal-8 是否影响 PA 角膜炎的结果： 调节中性粒细胞弹性蛋白酶和/或 TLR 途径。更好地表征 Gal-8 在发病机制中的作用 PA 角膜炎，在目标 3A 中，我们建议进行转录组和细胞因子/趋化因子蛋白质组分析 PA 感染的 WT 和 Gal-8 KO 小鼠角膜，以及 WT 和 Gal-8 KO 巨噬细胞和中性粒细胞。瞄准 3B，将进行研究以确定 Gal-8 是否可以靶向控制过度活跃的免疫反应 以及 PA 角膜炎小鼠模型中的角膜炎症。我们期望拟议的研究将提供 突破性的数据，不仅有助于了解PA角膜炎的分子机制，也有助于 开发基于半乳糖凝集素的新型疗法，可以治疗由以下原因引起的致盲免疫病理学： 细菌性角膜炎，以及其他眼部疾病，例如角膜移植排斥和干眼病。

项目成果

Galectin-8 Downmodulates TLR4 Activation and Impairs Bacterial Clearance in a Mouse Model of Pseudomonas aeruginosa Keratitis.

Galectin-8 下调铜绿假单胞菌角膜炎小鼠模型中 TLR4 激活并损害细菌清除。

• DOI: 10.4049/jimmunol.2200706
• 发表时间: 2023
• 期刊: Journal of immunology (Baltimore, Md. : 1950)
• 作者: Ramadan,Abdulraouf;Cao,Zhiyi;Hassan,Mujtaba;Zetterberg,Fredrik;Nilsson,UlfJ;Gadjeva,Mihaela;Rathinam,Vijay;Panjwani,Noorjahan
• 通讯作者: Panjwani,Noorjahan