An RNAi Screen for Genes that Modulate Retinal Ganglion Cell Survival

RNAi 筛选调节视网膜神经节细胞存活的基因

基本信息

批准号：
8539490
负责人：
Derek Stuart Welsbie
金额：
$ 15.39万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-09-01 至 2014-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8539490
关键词：
Adverse effects Biological Assay Blindness Brain Candidate Disease Gene Cause of Death Cell Culture Techniques Cell Death Cell Survival Cessation of life Clinic Complement Crush Injury Dependovirus Development Disease Dominant-Negative Mutation Dose Drug Targeting Evaluation Eye Eyedrops Future Genes Genome Glaucoma Government Health Image In Vitro Laboratories Lasers Lead Libraries MAP Kinase Kinase Kinase Mediating Mitogen-Activated Protein Kinase Kinases Modeling Molecular Mus N-terminal Nerve Crush Nerve Degeneration Neurodegenerative Disorders Neurons Neuroprotective Agents Operative Surgical Procedures Optic Nerve Optic Nerve Injuries Pathway interactions Persons Pharmaceutical Preparations Phosphotransferases Physiologic Intraocular Pressure Plasmids Protocols documentation RNA Interference RNA library Research Proposals Retina Retinal Ganglion Cells Rodent Model Role Signal Transduction Small Interfering RNA System Technology Technology Transfer Testing Therapeutic Agents Universities Viral Vector Vision Work adeno-associated viral vector base functional genomics gene function genome-wide in vitro testing in vivo inhibitor/antagonist kinase inhibitor member neuron loss neuroprotection novel optic nerve disorder prevent screening small hairpin RNA small molecule libraries stress-activated protein kinase 1 success treatment strategy

项目摘要

DESCRIPTION (provided by applicant): Pr Glaucoma is a neurodegenerative disease in which there is specific loss of retinal ganglion cells (RGCs). Current management is directed at lowering intraocular pressure (IOP), through the use of eye drops, laser treatment, and/or operative surgery. Although such treatment can be effective, sufficient IOP lowering cannot always be safely achieved, and sometime even with significant IOP lowering there still can be progression of optic nerve damage. In an effort to complement IOP-based therapy, efforts have been made to develop neuroprotective therapies that directly act to preserve RGC health and function. However, despite important laboratory advances, neuroprotection-based treatment approaches for glaucoma have not yet made it to the clinic. In order to help advance toward a clinically viable neuroprotective strategy, we have been pursuing a variety of approaches to identify novel drug targets and molecular pathways that could be used to promote RGC survival in glaucoma and other optic neuropathies. In a preliminary set of studies we have demonstrated the feasibility of performing an RNAi screen with primary cultures of murine RGCs to identify genes that modulate RGC survival. In this R21 application we propose to further optimize this technology, use it to perform a whole genome siRNA screen, and then validate the identified hits by testing their activity in vivo using an optic nerve crush model of optic nerve injury.

描述（由申请人提供）：青光眼是一种神经退行性疾病，其中视网膜神经节细胞（RGC）出现特定损失。目前的治疗方法是通过使用滴眼剂、激光治疗和/或手术来降低眼压（IOP）。尽管这种治疗可能有效，但并不总是能够安全地实现足够的眼压降低，有时即使显着降低眼压，视神经损伤仍然可能进展。为了补充基于 IOP 的治疗，人们正在努力开发直接保护 RGC 健康和功能的神经保护疗法。然而，尽管实验室取得了重要进展，但基于神经保护的青光眼治疗方法尚未进入临床。为了帮助实现临床上可行的神经保护策略，我们一直在寻求各种方法来确定可用于促进青光眼和其他视神经病变中 RGC 存活的新药物靶点和分子途径。在一组初步研究中，我们证明了利用小鼠 RGC 原代培养物进行 RNAi 筛选以确定调节 RGC 存活的基因的可行性。在这个 R21 应用中，我们建议进一步优化该技术，用它来执行全基因组 siRNA 筛选，然后通过使用视神经损伤的视神经挤压模型测试其体内活性来验证已识别的命中。