Human Ocular Surface Electrophysiology

项目总结 这是一个为期5年的K08导师奖，旨在促进Neel Pasricha博士的过渡，他是一名眼科医生， 角膜分专业培训，以独立调查员进行眼表眼部翻译研究。一个 加州大学旧金山分校经验丰富的导师、科学顾问和教学工作将帮助帕斯里查博士 研究技能传授，包括眼表电生理学、人角膜上皮细胞培训 培养模型，小分子药物发现的高通量筛选，实验小鼠模型， 和人类实验对象的临床研究。 这项研究的重点是干眼病(DED)，这是眼科尚未得到满足的主要需求，其特点是 伴有眼部症状的泪膜平衡受损。目前只有四个FDA批准的 DED的治疗方法，每一种都只针对炎症途径，疗效有限。眼科 表面由角膜和结膜组成，由表达离子转运的层状上皮细胞组成。 促进主动液体分泌或吸收的蛋白质，以调节泪液体积和渗透压。目标是 这项拟议的研究的目的是发现和推进促进泪液分泌的候选药物 通过眼球表面衬里的上皮细胞。 这项研究利用了一种新的眼表面电位差(OSPD)方法，该方法被引入到动物研究中 并在我在加州大学旧金山分校实习期间先进用于人类。OSPD测量电势 顶膜和基底膜离子转运体在上皮细胞间产生差异。在AIM 1中，体内 小鼠的OSPD测量将调查离子转运体在眼表液体运输中的作用， 特别关注氯化物和钾离子通道。AIM 2将使用体外高通量筛查 原代培养人角膜上皮细胞和体内实验小鼠研究 靶向钙激活的氯离子通道以增加泪液分泌的候选药物治疗 戴德。AIM 3将使用健康成年人的活体人体OSPD测量来测试一种促分泌药物 激活CFTR氯通道的2期临床试验的候选对象。长期的职业生涯 发展目标是建立一个强有力的跨学科研究计划，以推动 对眼表离子传输的基本了解并将其转化为小说 眼表疾病的诊断和治疗策略，包括DED。

PROJECT SUMMARY This is a 5-year mentored K08 award to facilitate the transition of Dr. Neel Pasricha, an ophthalmologist with cornea subspecialty training, to an independent investigator in ocular surface eye translational research. A strong group of experienced mentors at UCSF, scientific advisors, and didactic work will assist Dr. Pasricha’s research skills transfer, including training in ocular surface electrophysiology, human corneal epithelial cell culture models, high-throughput screening for small-molecule drug discovery, experimental mouse models, and human subject clinical studies. The research is focused on dry eye disease (DED), a major unmet need in ophthalmology characterized by impaired tear film homeostasis accompanied by ocular symptoms. There are currently just four FDA-approved therapies for DED, each targeting only the inflammatory pathway and having limited efficacy. The ocular surface, comprised of the cornea and conjunctiva, is lined by stratified epithelial cells expressing ion transport proteins that facilitate active fluid secretion or absorption to regulate tear fluid volume and osmolarity. The goal of the proposed research is to discover and advance drug candidates to promote tear fluid secretion by epithelial cells lining the ocular surface. This research utilizes a novel ocular surface potential difference (OSPD) method introduced in animal studies and advanced for use in humans during my residency at UCSF. OSPD measures the electrical potential difference generated across epithelia from apical and basal membrane ion transporters. In Aim 1, in vivo OSPD measurements in mice will investigate the role of ion transporters in ocular surface fluid transport, with particular focus on chloride and potassium channels. Aim 2 will use ex vivo high-throughput screening in primary human corneal epithelial cell cultures and in vivo experimental mouse studies to advance potential drug candidates that target calcium-activated chloride channels to increase tear fluid secretion for treatment of DED. Aim 3 will use in vivo human OSPD measurements in healthy adults to test a pro-secretory drug candidate in phase 2 clinical trial that activates the CFTR chloride channel. The long-term career development goal is to build a robust cross-disciplinary research program that advances the fundamental understanding of ocular surface ion transport and translates that knowledge into novel diagnostic and therapeutic strategies for ocular surface diseases, including DED.