Rhythmic Stimulation as a Therapeutic Option for Injured Retinal Ganglion Cells

节律刺激作为受损视网膜神经节细胞的治疗选择

基本信息

批准号：
8516361
负责人：
gustavo c munguba
金额：
$ 4.72万
依托单位：
UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
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：
2012-08-01 至 2016-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8516361
关键词：
Address Adult Affect African American Age-Years Axon Blindness Brain-Derived Neurotrophic Factor Cell Death Cell Survival Chimeric Proteins Chlamydomonas reinhardtii Cholera Toxin Cyclic AMP Development Disease Distal Electric Stimulation Functional disorder Genetic Transcription Glaucoma Glutamates Goals Green Algae Healthcare Hispanic Americans In Vitro Incidence Individual Injury Intracellular Second Messenger Label Lasers Light Minority Modality Nerve Crush Neurites Neurons Ophthalmoscopy Optic Nerve Optic Nerve Injuries Optical Coherence Tomography Pathway interactions Patients Physicians Physiologic Intraocular Pressure Play Population Proteins Proteomics Protons Retina Retinal Retinal Ganglion Cells Risk Factors Role Scanning Second Messenger Systems Site Testing Therapeutic Transgenic Mice axon regeneration in vivo inhibitor/antagonist injured insight light gated meetings modifiable risk neuroprotection neurotrophic factor novel therapeutics protein expression research study tool

项目摘要

DESCRIPTION (provided by applicant): Glaucoma is the second leading cause of irreversible blindness worldwide and it is estimated that 79.6 million people throughout the world will have glaucoma by 2020. Currently, increased intraocular pressure is the only known modifiable risk factor in glaucoma pathophysiology, and therefore the only treatment option available to patients and their physicians. Here we propose to investigate rhythmic stimulation of injured adult retinal ganglion cells (RGCs) as a novel therapeutic option in glaucoma. Our central hypothesis is that replication of the rhythmic RGC activity observed during development (retinal waves) will promote survival and axon regeneration of injured adult RGCs. To test this hypothesis we will specifically stimulate RGCs expressing a light-gated proton channel from the green algae Chlamydomonas reinhardtii. The Channelrhodopsin2 protein is expressed exclusively by RGCs of Thy1- COP4/EYFP line 9 (ChR2-YFP) transgenic mice. Using RGC specific Thy-1 expression of ChR2-YFP fusion protein allows us to both stimulate and track RGC survival overtime in vivo and in vitro. In specific aim 1 we will use a proteomics approach to assess whether in vivo rhythmic stimulation of RGCs modulates the expression of proteins involved in neurotrophic pathways. In specific aim 2, we will determine whether light induced rhythmic stimulation promotes cAMP independent RGC survival and neurite outgrowth in vitro by culturing RGCs in media containing cAMP inhibitors. In aim 3a we will determine whether rhythmic stimulation promotes RGC survival following optic nerve injury. Lastly, in aim 3b we will assess whether rhythmic stimulation promotes axon regeneration following optic nerve crush. The goal of this proposal is to explore a clinically applicable treatment modality to aid adult RGC neuron survival and axon regeneration after injury.

描述（由申请人提供）：青光眼是全球不可逆性失明的第二大原因，预计到 2020 年，全球将有 7960 万人患有青光眼。目前，眼内压升高是青光眼病理生理学中唯一已知的可改变的危险因素，因此，这是患者及其医生可用的唯一治疗选择。在这里，我们建议研究对受损成人视网膜神经节细胞（RGC）的节律性刺激作为青光眼的一种新的治疗选择。我们的中心假设是，在发育过程中观察到的节律性 RGC 活动（视网膜波）的复制将促进受损成年 RGC 的存活和轴突再生。为了验证这一假设，我们将专门刺激表达绿藻莱茵衣藻的光门控质子通道的 RGC。视紫红质通道蛋白 2 蛋白仅由 Thy1-COP4/EYFP 系 9 (ChR2-YFP) 转基因小鼠的 RGC 表达。使用 ChR2-YFP 融合蛋白的 RGC 特异性 Thy-1 表达使我们能够刺激和跟踪 RGC 在体内和体外的存活情况。在具体目标 1 中，我们将使用蛋白质组学方法来评估 RGC 的体内节律性刺激是否调节神经营养途径中涉及的蛋白质的表达。在具体目标 2 中，我们将通过在含有 cAMP 抑制剂的培养基中培养 RGC，确定光诱导的节律刺激是否能促进不依赖于 cAMP 的 RGC 存活和体外神经突生长。在目标 3a 中，我们将确定节律性刺激是否促进视神经损伤后 RGC 的存活。最后，在目标 3b 中，我们将评估节律性刺激是否促进视神经挤压后轴突再生。该提案的目标是探索一种临床适用的治疗方式来帮助成人 RGC 损伤后神经元存活和轴突再生。