Ocular surface functions of SLURP1

SLURP1的眼表功能

• 批准号: 10436949
• 负责人: Shivalingappa Kottur Swamynathan
• 金额: $ 22.93万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2023-01-03
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10436949
• 关键词: Address; Anti-Inflammatory Agents; Behavior; Cauterize; Chemotaxis; Cornea; Corneal Neovascularization; Development; Diagnostic; Disease; Dry Eye Syndromes; Endothelial Cells; Endothelium; Epithelial Cell Junction; Extravasation; Feedback; Fibrosis; Foundations; Healthcare Systems; Homeostasis; Human; Immune; In Vitro; Infiltration; Inflammation; Inflammatory; Injury; Intercellular Junctions; Knock-out; Knockout Mice; Knowledge; Laboratories; Liquid substance; Mediating; Mission; Mus; Nature; Neutrophil Infiltration; Ophthalmologist; Organism; Outcome; Outpatients; PI3K/AKT; Pathway interactions; Patients; Plasma; Production; Protein Family; Proteins; Publications; Reporting; Role; Severities; Signal Transduction; Silver Nitrate; Solid; TNF gene; Testing; Transforming Growth Factor beta; Tube; Umbilical vein; United States National Institutes of Health; Urokinase; Vascular Endothelial Cell; Visit; corneal epithelium; cytokine; experimental study; immunoregulation; member; monolayer; mouse model; neovascularization; neutrophil; new therapeutic target; novel therapeutics; ocular surface; public health relevance; therapeutic target; transcriptome; transcriptome sequencing

Summary The persistent gap in our understanding of ocular surface immunomodulation is a barrier for developing new therapies for inflammatory disorders that are responsible for a bulk of outpatient visits to an ophthalmologist. Previous publications from our laboratory established that the secreted Ly6/uPAR related protein-1 (SLURP1), a member of the Ly6 family of proteins is an immunomodulatory molecule at the ocular surface that: (i) is highly expressed in the corneal epithelium and secreted to the tear fluid; (ii) acts as a soluble scavenger of urokinase- type plasminogen activator (uPA); (iii) inhibits human umbilical vein endothelial cell (HUVEC) tube formation; (iv) suppresses neutrophil chemotaxis and transmigration through confluent endothelial monolayer in vitro; and (v) stabilizes epithelial cell junctions and suppresses TNF-α-induced cytokine production consistent with an anti-inflammatory function. Collectively, these studies identified SLURP1 as a potential therapeutic target for inflammatory disorders of the ocular surface. Here we propose to build upon these salient findings by testing the central hypothesis that ‘SLURP1 suppresses corneal angiogenic inflammation and neutrophil recruitment by regulating the TGF-β- and uPA-activities that promote NFκB-mediated production of pro-inflammatory molecules’. This hypothesis is supported by our prior publications described above, and exciting results from our unpublished preliminary studies wherein Slurp1 knockout (Slurp1X-/-) mouse corneas displayed dense corneal neovascularization and excessive neutrophil influx five days after silver nitrate cautery. Furthermore, RNA-Seq comparison of the naïve wild type and Slurp1X-/- mouse corneal transcriptomes identified key activators of angiogenic inflammation including TGF-β and NFκB pathway components to be upregulated in the absence of Slurp1, lending additional support for this hypothesis. We will test this hypothesis by employing mouse models and in vitro studies to pursue the following Specific Aims: Aim 1). Test the hypothesis that Slurp1 protects the cornea from undesirable angiogenic inflammation by suppressing unmitigated TGF-β and uPA activities that feed into NFκB pathway; Aim 2). Test the hypothesis that Slurp1 suppresses neutrophil influx into healthy corneas by promoting neutrophil maturation and clearance, and interfering with their extravasation; and Aim 3). Test the hypothesis that SLURP1 is negatively correlated with the severity of human dry eye disease and a useful therapeutic target for ocular surface inflammatory disorders. By elucidating promising new information related to the immunomodulatory functions of SLURP1, an abundantly expressed yet understudied protein, anticipated outcomes of this proposal directly address the NIH mission of ‘seeking fundamental knowledge about the nature and behavior of living systems’ and offer the potential for validating a novel therapeutic target for inflammatory disorders of the ocular surface that account for a significant burden on our healthcare system.

总结 我们对眼表免疫调节的理解上的持续差距是开发新的免疫调节的障碍。 炎症性疾病的治疗，这是导致大量门诊就诊于眼科医生的原因。 我们实验室以前的出版物确定分泌的Ly 6/uPAR相关蛋白-1（SLURP 1）， Ly 6蛋白家族的成员是眼表面的免疫调节分子，其：（i）高度 在角膜上皮中表达并分泌到泪液中;（ii）作为尿激酶的可溶性清除剂， 型纤溶酶原激活剂（uPA）;（iii）抑制人脐静脉内皮细胞（HUVEC）管形成; (iv)在体外抑制中性粒细胞的趋化性和通过汇合的内皮单层的迁移;和 (v)稳定上皮细胞连接并抑制TNF-α诱导的细胞因子产生， 抗炎作用。总的来说，这些研究将SLURP 1确定为潜在的治疗靶点， 眼表面的炎性疾病。在这里，我们建议通过测试来建立这些突出的发现 SLURP 1抑制角膜血管生成性炎症和中性粒细胞募集的中心假设 通过调节TGF-β和uPA活性，促进NFκ B介导的促炎性细胞因子的产生， molecules '。这一假设得到了我们上述先前出版物的支持， 我们未发表的初步研究中，Slurp 1敲除（Slurp 1X-/-）小鼠角膜显示致密的 银银烧灼后5天角膜新生血管形成和过量中性粒细胞流入。此外，委员会认为， 未处理野生型和Slurp 1X-/-小鼠角膜转录组的RNA-Seq比较确定了关键 血管生成性炎症的激活剂，包括TGF-β和NFκB途径组分，在 Slurp 1的缺失，为这一假设提供了额外的支持。我们将通过使用 小鼠模型和体外研究，以实现以下特定目的：目的1）。测试的假设 Slurp 1通过抑制未减轻的TGF-β和TGF-β受体， NFκB通路的uPA活性;目的2）。测试Slurp 1抑制中性粒细胞的假设 通过促进中性粒细胞成熟和清除，并干扰它们的 （三）目标（三）。检验SLURP 1与以下严重程度负相关的假设： 人干眼症和眼表面炎性疾病有用治疗靶点。通过 阐明了与SLURP 1的免疫调节功能相关的有希望的新信息， 表达尚未充分研究的蛋白质，该提案的预期结果直接解决了NIH的使命， “寻求有关生命系统的性质和行为的基本知识”，并提供潜在的 验证眼表炎性疾病的新治疗靶点， 这对我们的医疗保健系统造成了巨大的负担。