Defining the Immune Cell Roles in Meibomian Gland Dysfunction in the Context of Ocular Surface Immune Diseases

摘要 炎症在眉毛腺功能障碍中是一个被严重低估的角色--这是一种常见的病理 负责产生泪膜脂质的眼皮皮脂腺的状况。腺体病理性阻塞 开口和中央导管，被称为“梗阻性”眉板腺功能障碍，常伴有 患者眼表炎症。此外，这种类型的障碍无处不在，甚至更多 侵袭性，在眼表免疫性疾病患者中。后者在这里被定义为眼表 与全身免疫紊乱相关的病理学，如特应性(过敏)、史蒂文斯-约翰逊综合征/毒性 表皮坏死松解症、干燥综合征、移植物抗宿主病、瘢痕性类天疱疮等。不过， 有关眉板腺功能障碍的确切免疫机制仍不清楚。因为没有药理作用 药物是眉毛腺功能障碍的适应症，迫切需要揭示相关的 眼表免疫性疾病患者的免疫反应。在上一个资金周期中， 在我们的慢性类过敏性眼睛模型中，我们显示了中性粒细胞在眉板腺功能障碍中的直接作用。 疾病。基于这项工作和目前的文献，我们的中心假设是中性粒细胞在 在不同形式的眼表免疫性疾病中驱动眉板腺功能障碍。在当前 建议，我们将：i)阐明致病性中性粒细胞导致眉板腺功能障碍的机制。 我们建立的慢性类过敏性眼病模型；ii)破译中性粒细胞在眉板腺中的作用 一种新的自发遗传小鼠眼表免疫病模型的功能障碍；以及iii)MAP 不同眼表免疫力患者泪腺功能障碍的泪液免疫细胞图谱 疾病实体。除了小鼠模型，我们还将利用我们未发表的单细胞rna-seq数据集 来自血液、结膜和泪液的免疫细胞来自我们的过敏性眼病小鼠模型，与 计算、遗传和药理学工具，以确定致病中性粒细胞的机制。对于人类来说 样本，我们建立了一个36色流式细胞仪小组，用于深度免疫分析患者泪液和 结合临床专业知识，使用详细的临床元数据对每个个体进行注释。其基本原理是 对于这个项目来说，识别关键的免疫细胞亚群和相关的致病细胞程序 这将提供一个强有力的科学框架来描述脑膜腺的作用 人类眉板腺功能障碍中的炎症，以及确定新的治疗靶点和诊断 免疫标记物支持临床领域的努力。因此，预计这些研究将于 通过扩大关于炎症作用的知识的广度和深度，产生重大影响， 特别是中性粒细胞，在眉毛腺功能障碍中，最终提供了新的治疗和治疗机会。 诊断这种普遍存在的情况。

ABSTRACT Inflammation is a vastly underappreciated player in meibomian gland dysfunction – a common pathological condition of eyelid sebaceous glands responsible for producing tear film lipids. Pathologic blockage of the gland orifice and central duct, referred to as “obstructive” meibomian gland dysfunction, is regularly accompanied by ocular surface inflammation in patients. Moreover, this type of obstruction is ubiquitous, and even more aggressive, in patients with ocular surface immune disease. The latter is defined here as ocular surface pathology associated with systemic immune disorders, such as atopy (allergy), Stevens-Johnson syndrome/toxic epidermal necrolysis, Sjogren’s syndrome, graft versus host disease, cicatricial pemphigoid, and others. Still, the exact immune mechanisms involved in meibomian gland dysfunction remain elusive. As no pharmacological agents are indicated for meibomian gland dysfunction, there is a critical need for uncovering the pertinent immune responses of this pathobiology in patients with ocular surface immune disease. In the last funding cycle, we showed a direct role for neutrophils in meibomian gland dysfunction in our model of chronic-like allergic eye disease. Based on this work and the current literature, our central hypothesis is that neutrophils play a role in driving meibomian gland dysfunction across different forms of ocular surface immune diseases. In the current proposal, we will: i) elucidate the pathogenic neutrophil mechanisms that drive meibomian gland dysfunction in our established model of chronic-like allergic eye disease; ii) decipher the role of neutrophils in meibomian gland dysfunction in a novel and spontaneous genetic mouse model of ocular surface immune disease; and iii) map the tear immune cell landscape of meibomian gland dysfunction in patients with different ocular surface immune disease entities. In addition to mouse models, we will leverage our unpublished single cell RNA-seq dataset of immune cells from blood, conjunctiva, and tears from our allergic eye disease mouse model, paired with computational, genetic, and pharmacologic tools to pinpoint the pathogenic neutrophil mechanisms. For human samples, we established a 36-color flow cytometry panel for deep immune profiling of patient tears and have incorporated clinical expertise for the annotation of each individual with detailed clinical metadata. The rationale for this project is that identifying the critical immune cell subsets and associated pathogenic cellular programs that drive meibomian gland dysfunction will offer a strong scientific framework by which to delineate the role of inflammation in human meibomian gland dysfunction, as well as identify novel therapeutic targets and diagnostic immune markers to support efforts in the clinical space. Completion of these studies is therefore expected to have a significant impact by expanding the breadth and depth of knowledge regarding the role of inflammation, specifically neutrophils, in meibomian gland dysfunction, ultimately providing new opportunities to treat and diagnose this pervasive condition.