Contribution of mast cells in non-allergic ocular inflammation

肥大细胞在非过敏性眼部炎症中的作用

基本信息

批准号：
10164794
负责人：
Sunil K Chauhan
金额：
$ 47.77万
依托单位：
SCHEPENS EYE RESEARCH INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-09-01 至 2023-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10164794
关键词：
Adrenal Cortex Hormones Adverse effects Age related macular degeneration Allergic Disease Anti-Inflammatory Agents Architecture Autoimmunity Biological Assay Blindness Blood Circulation CXCL2 gene Cell physiology Chemotactic Factors Chemotaxis Cornea Corneal Injury Data Disease Effector Cell Elements Epithelial Cells Eye Eye Injuries Fibroblasts Foundations Goals Graft Rejection HMGB1 gene High Prevalence Hour IgE Immune Immunology procedure Immunosuppressive Agents Impaired wound healing In Vitro Infection Infiltration Inflammation Inflammation Mediators Inflammatory Injury Innate Immune Response Interleukin-1 beta Keratoplasty Knockout Mice Laboratories Light Malignant Neoplasms Mediating Mediator of activation protein Molecular Mus Necrosis Neutrophil Activation Neutrophil Infiltration Neutrophilic Infiltrate Pathogenicity Pathology Pharmaceutical Preparations Phase Play Publications Publishing Reporting Research Role Sentinel Series Site Sterility Testing Therapeutic Tissues Treatment Efficacy Tryptase Visual impairment Wound Infection allergic response base beta-n-acetylhexosaminidase chemokine corneal epithelium cytokine design differential expression experimental study immune activation immune function in vivo injured interest macrophage mast cell migration mouse model neutrophil novel novel therapeutic intervention novel therapeutics prevent receptor reconstitution recruit repository response sensor side effect tissue injury tissue repair translational impact

项目摘要

Ocular injury is a leading cause of corneal blindness, resulting in millions of cases of visual impairment globally each year. Currently, the uncontrolled immune activation and tissue damage that occur following ocular injury are treated with non-specific anti-inflammatory drugs (e.g. corticosteroids), which are rife with deleterious side effects such as delayed wound healing and infection. Our research aims to identify the specific cellular and molecular factors that mediate early immune cell activation and infiltration following ocular injury, so that novel therapeutics may be developed. Our group’s recently published reports and preliminary data have identified tissue-resident mast cells as the primary reservoir of neutrophil-chemoattractants at the cornea, and have established that instant release of preformed CXCL2 by mast cells is critical for early neutrophil migration (<1 hour) following injury. Early-recruited ‘scout’ neutrophils subsequently release chemoattractant factors that drive a robust second phase of neutrophil infiltration, amplifying the innate immune response. Excessive recruitment and activation of neutrophils is known to cause deleterious inflammation and damage corneal architecture. These observations pose a critical question: How do mast cells sense tissue injury and promote early neutrophil recruitment? Our data show that damaged corneal epithelial cells (but not resident stromal fibroblasts or macrophages) release inflammatory mediators that activate mast cells. In accordance with our laboratory’s expertise in immunological assays and utilizing a well-characterized murine model of sterile corneal stromal injury, we propose a series of novel experiments to decipher the role of mast cells as sensors of tissue injury. In Aim 1, we will test the hypothesis that danger-associated inflammatory molecules IL33, IL36γ and HMGB1 released from corneal epithelial cells activate mast cells in an IgE- independent manner. Specifically, we will (i) establish that necrotic epithelial cells stimulate mast cells relative to healthy and IL1β-treated epithelial cells using mast cell-specific tryptase and β-hexosaminidase release assays; and (ii) identify the key danger-associated molecules expressed by corneal epithelial cells that activate mast cells. In Aim 2, we will test the hypothesis that in vivo blockade of IL36γ function will be more effective, relative to IL33 and HMGB1, in suppressing mast cell activation, resulting in reduced neutrophil infiltration and tissue damage. Specifically, we will determine the effect of (i) silencing select mast cell- activating mediators, and (ii) silencing receptors for select mediators on mast cells, on mast cell activation and early neutrophil recruitment during corneal injury. We will also evaluate the therapeutic potential of local blockade of select mediators. It is anticipated that this research will have significant translational impact due to the high prevalence of ocular injury and inflammatory disease, as well as the relevance of mechanisms governing neutrophil infiltration to non-ocular tissues.

眼部损伤是角膜失明的主要原因，导致全球视力障碍数百万每年。目前，眼部损伤后发生的不受控制的免疫激活和组织损伤用非特异性抗炎药（例如皮质类固醇）治疗，这些药物充满有害的一面诸如延迟伤口愈合和感染之类的影响。我们的研究旨在确定特定的细胞和眼部损伤后培养早期免疫细胞激活和浸润的分子因素，因此可以开发治疗。我们小组最近发布的报告和初步数据已经确定了组织居民桅杆细胞为角膜上嗜中性粒细胞折射剂的主要储层，并确定肥大细胞即时释放预成式的CXCL2对于早期嗜中性粒细胞迁移至关重要（<1小时）受伤。早期收集的“侦察”中性粒细胞随后释放出促进稳健的趋化因子中性粒细胞浸润的第二阶段，扩大了先天免疫反应。过度招募和已知中性粒细胞的激活会引起有害的炎症和损害角膜结构。这些观察结果提出了一个关键的问题：桅杆细胞如何感受组织损伤并促进早期嗜中性粒细胞招募？我们的数据表明，角膜上皮细胞受损（但没有居民成纤维细胞或巨噬细胞）释放激活肥大细胞的炎症介质。按照我们的实验室在免疫学测定方面的专业知识，并使用无菌特征的鼠模型角膜基质损伤，我们提出了一系列新型实验，以破译肥大细胞的作用组织损伤的传感器。在AIM 1中，我们将测试与危险相关的炎症的假设分子IL33，IL36γ和HMGB1从角膜上皮细胞中释放出来激活IgE-中的肥大细胞独立的方式。具体而言，我们将（i）确定坏死性上皮细胞刺激肥大细胞相对于使用肥大细胞特异性胰蛋白酶和β-己糖胺酶的健康和IL1β处理的上皮细胞发布测定；（ii）确定角膜上皮细胞表达的关键危险相关分子激活肥大细胞。在AIM 2中，我们将测试以下假设：IL36γ函数的体内阻滞相对于IL33和HMGB1，在抑制肥大细胞激活方面更有效，导致中性粒细胞降低浸润和组织损伤。具体而言，我们将确定（i）沉默选择肥大细胞的影响激活介质的介体在肥大细胞激活上的肥大细胞上精选介质的受体沉默。和角膜损伤期间的早期嗜中性粒细胞募集。我们还将评估本地的治疗潜力封锁选定的调解人。预计这项研究将产生重大的翻译影响由于眼部损伤和炎症性疾病的高流行以及机制的相关性管理中性粒细胞浸润到非眼组织。