Regulation of Corneal Inflammation by Fas Ligand

描述(由申请者提供)：我获得临床科学家发展指导奖的最终目标是获得先天免疫领域的专业知识。具体地说，我将研究Fas配体(FasL)在调节角膜中性粒细胞介导的(天然免疫)炎症中的作用。这次培训与我之前在适应性免疫方面的研究经验相结合，将使我有机会制定一个全面的研究计划来研究角膜免疫学。科尔眼科研究所和克利夫兰临床基金会的免疫学部门提供必要的临床和基础科学支持，使临床科学家的发展和从事转化医学的独特机会得以实现。总体而言，该机构的培训环境将为我提供必要的技术和智力工具，使我成为一名成功的视觉研究研究员。我的长期职业目标是建立一个全面和独立的研究计划，致力于剖析角膜用于控制炎症的调节免疫机制。这项研究产生的科学发现将用于开发针对目标的免疫疗法，用于治疗破坏性的角膜炎症性疾病。本项目的目的是验证这一新的假设，即角膜中表达的FasL的形式是炎症差异调节的关键因素：可溶性FasL块和膜FasL诱导角膜炎。初步研究表明，可溶性FasL通过抑制中性粒细胞活化而发挥抗炎作用，而膜FasL则通过激活中性粒细胞而起到促炎作用。本研究将实现一种在体转染腺病毒载体的方法，在角膜组织中表达可溶性和膜型FasL，并确定角膜组织中性粒细胞激活的生物调节。利用分离的中性粒细胞和转染膜和可溶性FasL的细胞系进行的体外实验将补充这些研究，并将有助于剖析由这些分子差异调控的细胞和分子信号。此外，我们还将利用体内转染技术，在一个具有临床意义的脂多糖性角膜炎模型中，确定可溶性Fas-L在下调和预防中性粒细胞介导的眼损伤中的作用。我们期望这些研究将提供关于FasL调节眼部炎症的新信息，并导致使用可溶性FasL治疗和预防中性粒细胞介导的眼损伤的新疗法的发展。

DESCRIPTION (provided by applicant): My ultimate goal for the mentored clinical scientist development award is to acquire expertise in the field of innate immunity. Specifically, I will investigate the role of Fas Ligand (FasL) in regulating neutrophil-mediated (innate immunity) inflammation in the cornea. The combination of this training with my previous research experience in adaptive immunity will give me the opportunity to develop a comprehensive research program to study corneal immunology. The Cole Eye Institute and the Immunology Department at the Cleveland Clinic Foundation provide the clinical and basic science support necessary to allow the development of clinical scientists and the unique opportunity to practice translational medicine. Overall, the training environment, at this institution will provide me with both the technical and intellectual tools necessary to become a successful investigator in vision research. My long-term career goal is to establish a comprehensive and independent research program dedicated to dissect the regulatory immune mechanism used by the cornea to control inflammation. Scientific findings generated form this research will be used to development target-specific immunotherapies for the treatment of destructive inflammatory diseases of the cornea. The objective of this project is to test the novel hypothesis that the forms of FasL expressed in the cornea are a key factor in the differential regulation of inflammation: soluble FasL blocks and membrane FasL induces keratitis. Preliminary data demonstrate that soluble FasL is anti-inflammatory by inhibiting neutrophil activation, while membrane FasL activates them and is pro-inflammatory. The proposed studies will implement a novel method of in vivo transfection of corneal stromal cells using adenoviral vectors to express soluble and membrane FasL in the cornea and determine the biological regulation of neutrophil activation in corneal tissue. In vitro experiments using isolated neutrophils and cells lines transfected with membrane and soluble FasL will complement these studies and will facilitate the dissection of cellular and molecular signals differentially regulated by these molecules. Moreover, we will use the in vivo transfection technique to determine the role of soluble Fas L in the down-regulation and prevention of neutrophil-mediated ocular damage in a model of lipopolysaccharide induced keratitis that has clinical implications. We anticipate that these studies will yield new information about the regulation of ocular inflammation by FasL and lead to the development of new treatments using soluble FasL to treat and prevent neutrophil-mediated ocular damage.