An RNAi Screen for Genes that Modulate Retinal Ganglion Cell Survival

描述(申请人提供)：PR青光眼是一种神经退行性疾病，有视网膜神经节细胞(RGC)的特异性丢失。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。虽然这种治疗方法可以有效，但不能总是安全地达到足够的眼压降低，有时即使眼压显著降低，视神经损伤仍可能进展。为了补充基于眼压的治疗，已经努力开发神经保护疗法，直接作用于保护RGC的健康和功能。然而，尽管取得了重要的实验室进展，基于神经保护的青光眼治疗方法尚未进入临床。为了促进临床上可行的神经保护策略的发展，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，以促进青光眼和其他视神经疾病的RGC生存。在一组初步的研究中，我们已经证明了对小鼠视网膜节细胞的原代培养进行RNAi筛选以识别调节视网膜节细胞存活的基因的可行性。在这项R21应用中，我们建议进一步优化这项技术，使用它进行全基因组siRNA筛选，然后通过使用视神经损伤的视神经挤压模型在体内测试它们的活性来验证识别的HITS。

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.