Regulation of Corneal Inflammation by Fas Ligand

描述（由申请人提供）：我的指导临床科学家发展奖的最终目标是获得先天免疫领域的专业知识。 具体来说，我将研究Fas配体（FasL）在调节角膜中嗜酸性粒细胞介导的（先天免疫）炎症中的作用。 这次培训与我以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。科尔眼科研究所和免疫学系在克利夫兰诊所基金会提供必要的临床和基础科学支持，使临床科学家的发展和实践转化医学的独特机会。 总的来说，这个机构的培训环境将为我提供必要的技术和知识工具，使我成为一名成功的视觉研究调查员。 我的长期职业目标是建立一个全面而独立的研究项目，致力于剖析角膜用于控制炎症的调节免疫机制。 从这项研究中产生的科学发现将用于开发靶向特异性免疫疗法，用于治疗角膜的破坏性炎症疾病。 本项目的目的是检验新的假设，即角膜中表达的FasL形式是炎症差异调节的关键因素：可溶性FasL阻滞剂和膜FasL诱导角膜炎。 初步数据表明，可溶性FasL是通过抑制中性粒细胞活化抗炎，而膜FasL激活它们，是促炎。 拟议的研究将实施一种新的方法，使用腺病毒载体在角膜中表达可溶性和膜FasL的角膜基质细胞的体内转染，并确定在角膜组织中的中性粒细胞活化的生物调节。 使用分离的中性粒细胞和细胞系转染膜和可溶性FasL的体外实验将补充这些研究，并将促进这些分子差异调节的细胞和分子信号的解剖。 此外，我们将使用在体内转染技术，以确定可溶性Fas L的作用，在下调和预防嗜中性粒细胞介导的眼损伤的模型中的脂多糖诱导的角膜炎，具有临床意义。我们预计，这些研究将产生新的信息调节眼部炎症的FasL，并导致新的治疗方法的发展，使用可溶性FasL治疗和预防嗜酸性粒细胞介导的眼损伤。

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.