Analysis of chemical modulators for corneal endothelial dystrophies

角膜内皮营养不良的化学调节剂分析

• 批准号: 10661788
• 负责人: Matilda F Chan
• 金额: $ 40.38万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-30 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10661788
• 关键词: Address; Anaphylaxis; Apoptosis; Bilateral; Binding; Biological Assay; Blindness; CRISPR interference; Carrier Proteins; Cell Survival; Cell physiology; Cells; Chemicals; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Confocal Microscopy; Cornea; Corneal Diseases; Corneal Endothelium; Corneal dystrophy; Crystallins; Data; Defect; Disease; Endothelium; Equilibrium; Eye; Eye diseases; Free Energy; Fuchs' Endothelial Dystrophy; Functional disorder; Genes; Genetic Screening; Glafenine; Goals; In Vitro; Inflammatory; Inherited; Investigation; Keratoplasty; Kidney; Lead; Length; Libraries; Link; Mass Spectrum Analysis; Measurement; Measures; Medical; Methods; Microscopy; Mutation; Non-Steroidal Anti-Inflammatory Agents; Open Reading Frames; Osmosis; Outcome; Patients; Phenotype; Point Mutation; Process; Proteins; Quality Control; Research; Research Proposals; Series; Site; Structure-Activity Relationship; Surface Plasmon Resonance; Syndrome; Testing; Therapeutic; Toxic effect; Toxicity Tests; Visual impairment; alpha-Crystallins; analog; candidate identification; chemical genetics; clinical candidate; congenital cataract; cytotoxic; cytotoxicity; early onset; experimental study; functional gain; functional restoration; genome-wide; high throughput screening; high throughput technology; improved; in vitro Assay; insight; loss of function; misfolded protein; mutant; novel; novel therapeutic intervention; preclinical development; protein folding; protein function; protein misfolding; repaired; response; screening; sight restoration; small molecule; small molecule libraries; trafficking

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.

项目摘要 角膜内皮营养不良是视力丧失的常见原因，其特征在于缓慢的 进行性双侧角膜内皮功能障碍。恢复视力的主要治疗选择是 这些病人仍然接受角膜移植，因为医疗管理通常不足。突变 SLC 4A 11基因与几种内皮营养不良有关，包括Fuchs内皮角膜 营养不良（FECD）、先天性遗传性角膜内皮营养不良（CHED）和Harboyan综合征。 SLC 4A 11是一种转运蛋白，其功能是维持角膜内皮中的渗透平衡，并且许多 在疾病中发现的SLC 4A 11的点突变导致全长蛋白质的错误折叠。最近有 有证据表明，帮助突变体SLC 4A 11重新获得正确的折叠是一种有前途的治疗方法。现有 小规模，高通量化学筛选确定了非甾体抗炎药格拉非宁， 其纠正错误折叠的SLC 4A 11中的折叠缺陷的能力，以及在纠正后突变体SLC 4A 11 蛋白质恢复功能活性。尽管格拉非宁由于过敏反应而不是临床使用的候选者， 肾毒性，其疗效支持使用其他小分子折叠校正剂来修复缺陷的方法 角膜内皮营养不良中的SLC 4A 11。我们的长期目标是开发一流的治疗方法 用于因角膜中蛋白质错误折叠而导致视力受损的患者。我们打算检验这个假设 通过表型筛选试验鉴定的化合物将促进突变体的正确折叠， SLC 4A 11和恢复角膜内皮细胞功能。本提案的目标是：目标1）执行 聚焦筛选以鉴定SLC 4A 11折叠的校正剂;目的2）进行大的、无偏倚的化学文库 和遗传筛选，以确定新的目标和机制;和 非甾体抗炎药对角膜细胞的影响。利用先进的实验方法，包括高含量的显微镜为基础的 筛选试验和基于CRISPR的新型遗传筛选，拟议的研究将提供对 作为角膜内皮营养不良的治疗策略的蛋白质折叠缺陷的校正。结果 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导致视力丧失的患者： 角膜内皮营养不良，并建立新的角膜疾病治疗方法。