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：
8400753
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/8400753
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. PUBLIC HEALTH RELEVANCE: Glaucoma is the leading cause of irreversible blindness worldwide, affecting over 67 million people and up to 4% of individuals over 40 years of age. Disease incidence in the USA is disproportionately elevated in minority populations such as African Americans and Hispanic Americans and is responsible for nearly $3 billion (USD) in healthcare spending every year. 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 in vivo rhythmic stimulation of injured adult RGCs as a novel therapeutic option to support retinal ganglion survival in glaucoma.

描述（由申请人提供）：青光眼是全球不可逆转的失明的第二主要原因，据估计，到2020年，全世界有7,960万人将患有青光眼。目前，人眼内压力增加是唯一可在青光眼的病理生理学危险因素，因此，唯一可用于患者及其物理学的治疗方法。在这里，我们建议研究对受伤的成年视网膜神经节细胞（RGC）作为青光眼的一种新治疗选择的节奏刺激。我们的中心假设是，在发育过程中观察到的节奏RGC活性的复制（视网膜波）将促进受伤的成年RGC的存活和轴突再生。为了检验这一假设，我们将特别刺激从绿藻reinhardtii中表达光门控质子通道的RGC。 ChannelRhodopsin2蛋白仅由Thy1-COP4/EYFP第9行（CHR2-YFP）转基因小鼠的RGC仅表示。使用ChR2-YFP融合蛋白的RGC特异性THY-1表达，我们可以刺激和跟踪RGC在体内和体外的加班。在特定目标1中，我们将使用蛋白质组学方法来评估RGC的体内节律刺激是否调节涉及神经营养途径的蛋白质的表达。在特定的目标2中，我们将通过在含有CAMP抑制剂的培养基中培养RGC来确定光诱导的节奏刺激是否会在体外促进CAMP独立的RGC存活和神经突生长。在AIM 3A中，我们将确定节奏刺激是否促进视神经损伤后的RGC存活。最后，在AIM 3B中，我们将评估节奏刺激是否促进视神经挤压后的轴突再生。该建议的目的是探索一种临床适用的治疗方式，以帮助成人RGC 损伤后神经元存活和轴突再生。公共卫生相关性：青光眼是全世界不可逆失明的主要原因，影响了6700万人超过6700万人，最多40岁以上的个人。在美国的少数民族和西班牙裔美国人等少数民族人口中，美国的疾病发病率不成比例地升高，并且每年的医疗保健支出造成了近30亿美元（USD）。目前，升高的眼内压是青光眼病理生理学中唯一已知的可修改危险因素，因此是患者及其医生可用的唯一治疗选择。在这里，我们建议研究受伤的成年RGC作为支持视网膜神经节生存的新型治疗选择的体内节奏刺激。