An RNAi Screen for Genes that Modulate Retinal Ganglion Cell Survival

描述（由申请人提供）：Pr型青光眼是一种神经退行性疾病，其视网膜神经节细胞（RGCs）特异性丧失。目前的治疗方法是通过使用眼药水、激光治疗和/或手术来降低眼压。虽然这种治疗是有效的，但不能总是安全地达到充分的IOP降低，有时即使显著降低IOP，视神经损伤仍可能进展。为了补充以io为基础的治疗，人们努力开发神经保护疗法，直接保护RGC的健康和功能。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。为了帮助推进临床可行的神经保护策略，我们一直在寻求各种方法来确定新的药物靶点和分子途径，这些靶点和分子途径可用于促进青光眼和其他视神经病变的RGC存活。在一组初步研究中，我们已经证明了用小鼠RGC原代培养物进行RNAi筛选以鉴定调节RGC存活的基因的可行性。在本R21应用中，我们建议进一步优化该技术，使用它进行全基因组siRNA筛选，然后通过使用视神经损伤视神经挤压模型测试其在体内的活性来验证鉴定的命中。

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. PUBLIC HEALTH RELEVANCE: Vision loss from glaucoma is caused by the death of retinal ganglion cells (RGCs), the nerve cells in the retina that carry vision from the eye to the brain. We are using RNA interference-based approaches to try to better understand the signals that trigger RGCs to die. This work will hopefully identify new drug targets that could be used to reduce or prevent RGC death and vision loss, and thereby lead to novel treatment strategies for glaucoma and other optic neuropathies.