Analysis of chemical modulators for corneal endothelial dystrophies

项目总结 角膜内皮营养不良是导致视力丧失的常见原因，其特点是 进行性双侧角膜内皮功能障碍。恢复视力的主要治疗选择 这些患者仍然接受角膜移植，因为医疗管理通常不够充分。基因突变 SLC4A11基因与包括Fuchs内皮角膜在内的多种内皮细胞营养不良有关 营养不良(FECD)、先天性遗传性内皮角膜营养不良(CHED)和哈博扬综合征。 SLC4A11是一种转运蛋白，功能是维持角膜内皮的渗透平衡，以及许多 在疾病中发现的SLC4A11的点突变会导致全长蛋白质的错误折叠。有最近的 有证据表明，帮助突变的SLC4A11重新获得适当的折叠是一种很有前途的治疗方法。一位副院长 小规模高通量化学筛选鉴定非甾体抗炎药格列芬宁 其纠正错误折叠的SLC4A11中的折叠缺陷的能力，以及在纠正之后突变的SLC4A11 蛋白质恢复了功能活性。尽管格拉芬因过敏反应和 肾毒性，其有效性支持使用其他小分子折叠校正器修复缺陷的方法 SLC4A11在角膜内皮营养不良中的表达我们的长期目标是开发一流的治疗方法 适用于因角膜蛋白质错误折叠而导致视力障碍的患者。我们建议检验这一假设 通过表型筛选试验鉴定的化合物将促进突变的正确折叠 SLC4A11，恢复角膜内皮细胞功能。这项建议的目标是：目标1)执行 聚焦筛选以确定SLC4A11折叠的校正子；目标2)执行大型、无偏见的化学文库 和遗传筛选，以确定新的靶点和机制；以及目标3)评估 非甾体抗炎药对角膜细胞的影响。使用先进的实验方法，包括基于高含量显微镜的 通过筛查分析和基于CRISPR的新型基因筛查，拟议的研究将提供对 矫正蛋白质折叠缺陷作为治疗角膜内皮营养不良的策略。结果来自 这项研究将用于开发一种新的非手术治疗方案，用于治疗因以下原因而致视力丧失的患者 角膜内皮营养不良，并建立一种新的治疗角膜疾病的方法。

PROJECT SUMMARY Corneal endothelial dystrophies are a common cause of vision loss and are characterized by a slowly progressive, bilateral dysfunction of the corneal endothelium. The main therapeutic option to restore vision in these patients remains corneal transplantation, as medical management is usually inadequate. Mutations in the SLC4A11 gene are associated with several endothelial dystrophies, including Fuchs endothelial corneal dystrophy (FECD), congenital hereditary endothelial corneal dystrophy (CHED), and Harboyan syndrome. SLC4A11 is a transporter protein that functions to maintain osmotic balance in corneal endothelium, and many point mutations in SLC4A11 found in disease lead to misfolding of the full-length protein. There is recent evidence that assisting mutant SLC4A11 to regain proper folding is a promising therapeutic approach. A prior small-scale, high throughput chemical screen identified the non-steroidal anti-inflammatory drug glafenine for its ability to correct folding defects in misfolded SLC4A11, and following correction the mutant SLC4A11 protein regained functional activity. Though glafenine is not a candidate for clinical use due to anaphylaxis and renal toxicity, its efficacy supports an approach using other small molecule folding correctors to repair defective SLC4A11 in corneal endothelial dystrophies. Our long-term objective is to develop first-in-class therapeutics for patients with vision impairment due to protein misfolding in the cornea. We propose to test the hypothesis that compounds identified through phenotypic screening assays will promote the correct folding of mutant SLC4A11 and restore corneal endothelial cell function. The goals of this proposal are to: Aim 1) Perform focused screening to identify correctors of SLC4A11 folding; Aim 2) Perform large, unbiased chemical library and genetic screening to identify novel targets and mechanisms; and Aim 3) Assess cytotoxic effects of NSAIDs on corneal cells. Using advanced experimental methods, including high-content microscopy-based screening assays and novel CRISPR-based genetic screens, the proposed studies will provide insight into the correction of protein folding defects as a therapeutic strategy for corneal endothelial dystrophies. Results from the study will be used to develop a new, non-surgical treatment option for patients with vision loss due to corneal endothelial dystrophies and establish a novel therapeutic approach to corneal disease.