Novel Glaucoma Treatment Using Genome Editing

使用基因组编辑的新型青光眼治疗

基本信息

批准号：
9039605
负责人：
Abbot Frederick Clark
金额：
$ 50.84万
依托单位：
UNIVERSITY OF IOWA
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-04-01 至 2019-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9039605
关键词：
Address Affect Anterior Apoptosis Aqueous Humor Blindness Cell Death Cells Chronic Clustered Regularly Interspaced Short Palindromic Repeats DNA DNA Sequence Alteration Development Dexamethasone Diagnosis Disease Endoplasmic Reticulum Event Eye Eye diseases Failure Functional disorder Gene Mutation Gene Targeting Genes Genetic Glaucoma Goals Health Human Laboratories Lead Light Measures Methodology Methods Modality Modeling Molecular Chaperones Mus Mutation Ocular Hypertension Organ Culture Techniques Pathogenesis Pathology Pathway interactions Patients Perfusion Phenotype Physiologic Intraocular Pressure Play Primary Open Angle Glaucoma Proteins RNA Sequences Retinal Ganglion Cells Risk Factors Role Signaling Protein Site Steroids System Technology Testing Therapeutic Time Tissues Trabecular meshwork structure Transgenic Mice Work axonal degeneration base body system common treatment endoplasmic reticulum stress gain of function mutation gene function gene therapy genetic risk assessment genome editing improved in vivo insight mouse model mutant myocilin novel nuclease prevent response sensor success tool transcription factor

项目摘要

DESCRIPTION (provided by applicant): Primary open angle glaucoma (POAG) is a leading cause of irreversible vision loss and blindness. Current treatments of POAG are aimed at reducing intraocular pressure (IOP), the most important risk factor for the development and progression of the disorder. However, current glaucoma treatments do not address the underlying disease mechanisms. Mutations in the myocilin gene (MYOC) are the most common known genetic cause of glaucoma. Three developments provide us with an opportunity to investigate novel treatments of glaucoma directed at the underlying disease mechanisms: 1. The development of a POAG mouse model (Tg- MYOCY437H) in our laboratory that expresses mutant myocilin encoded by the MYOC gene; 2. The use of the Tg-MYOCY437H mouse model to demonstrate in vivo that mutant myocilin accumulates in the endoplasmic reticulum (ER) and induces ER stress in the trabecular meshwork (TM) of the eye as a key mechanistic component leading to glaucoma; and 3. The recent development of genome editing nucleases as a valuable tool for gene therapy applications. Furthermore, the fact that myocilin-associated POAG results from dominant gain of function mutations and the fact that ER stress induced apoptosis in the TM leads to POAG provide an opportunity to explore novel glaucoma treatments by in vivo elimination of mutant MYOC and/or apoptosis pathway genes (e.g. Chop). We propose to utilize the Tg-MYOCY437H mouse model of glaucoma to further explore the role of ER stress in POAG and to use a novel genome editing method (CRISPR) to explore novel POAG treatments, specifically the elimination of mutant myocilin from the TM of Tg-MYOCY437H mice, as well as the elimination of ER stress genes from TM cells. Clustered regularly interspaced short palindromic repeats (CRISPR) is an endogenous nuclease system, which uses RNA sequences to guide the cleavage of DNA by the CRISPR-associated nuclease Cas9. In addition, we will explore the role of ER stress in steroid induced glaucoma for the purpose of determining whether the genome editing strategy of glaucoma treatment can be applied to this form of glaucoma. Importantly, we will extend our work to humans by exploring the utility of genome editing in a human eye perfusion organ culture system. We believe that Cas9 can be used to remove the mutant MYOC gene and provide the first effective cure for MYOC mutation-induced glaucoma. In addition, the eye is an ideal test site for establishing the therapeutic potential of the Cas9 technology, and this work will pave the path for the application of Cas9 for treating a variety of genetic eye disorders, as well disorders affecting other organ systems.

描述（由申请人提供）：原发性开角型青光眼（POAG）是导致不可逆视力丧失和失明的主要原因。目前 POAG 的治疗旨在降低眼内压 (IOP)，眼内压是该疾病发生和进展的最重要的危险因素。然而，目前的青光眼治疗并没有解决潜在的疾病机制。肌纤蛋白基因 (MYOC) 突变是青光眼最常见的已知遗传原因。三项进展为我们提供了一个机会来研究针对潜在疾病机制的青光眼新疗法： 1. 在我们的实验室中开发了 POAG 小鼠模型（Tg-MYOCY437H），该模型表达由 MYOC 基因编码的突变肌纤蛋白； 2. 使用Tg-MYOCY437H小鼠模型在体内证明突变型肌纤蛋白在内质网（ER）中积累并诱导眼睛小梁网（TM）中的ER应激，作为导致青光眼的关键机制成分； 3. 基因组编辑核酸酶的最新发展作为基因治疗应用的有价值的工具。此外，肌纤蛋白相关的 POAG 是由功能突变的显性获得导致的，并且 ER 应激诱导 TM 细胞凋亡导致 POAG 的事实提供了通过体内消除突变型 MYOC 和/或细胞凋亡途径来探索新的青光眼治疗的机会基因（例如 Chop）。我们建议利用Tg-MYOCY437H青光眼小鼠模型进一步探讨ER应激在POAG中的作用，并使用新型基因组编辑方法（CRISPR）探索新的POAG治疗方法，特别是从Tg的TM中消除突变肌纤蛋白-MYOCY437H小鼠，以及从TM细胞中消除ER应激基因。成簇规则间隔短回文重复序列 (CRISPR) 是一种内源性核酸酶系统，它使用 RNA 序列引导 CRISPR 相关核酸酶 Cas9 对 DNA 进行切割。此外，我们将探讨ER应激在类固醇诱导的青光眼中的作用，以确定青光眼治疗的基因组编辑策略是否可以应用于这种形式的青光眼。重要的是，我们将通过探索基因组编辑在人眼灌注器官培养系统中的效用，将我们的工作扩展到人类。我们相信Cas9可以用来去除突变的MYOC基因，并为MYOC突变诱发的青光眼提供第一个有效的治疗方法。此外，眼睛是确定Cas9技术治疗潜力的理想测试部位，这项工作将为Cas9应用于治疗各种遗传性眼部疾病以及影响其他器官系统的疾病铺平道路。