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Sigma-1 Chaperone-Mediated in vivo Neuroprotection in the Retina

Sigma-1 伴侣介导的体内视网膜神经保护

基本信息

批准号：
8346582
负责人：
Lianwang Guo
金额：
$ 33.64万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
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 2017-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8346582
关键词：
Acute Apoptosis Apoptotic Blindness Cell Death Cell Transplants Cells Cessation of life Characteristics Chronic Data Degenerative Disorder Disease model Environment Event Exposure to Eye Eye diseases Genes Glaucoma Goals Human Investigation Knock-out Lead Light Mediating Membrane Proteins Methods Modeling Molecular Chaperones Morphology Mus Mutation Nerve Crush Nerve Degeneration Neurodegenerative Disorders Neuroprotective Agents Opsin Optic Nerve Oxidative Stress Oxygen Photoreceptors Population Process Property Proteins Reactive Oxygen Species Reporting Retina Retinal Retinal Degeneration Retinal Ganglion Cells Retinal Photoreceptors Retinitis Pigmentosa Role Stem cells Subfamily lentivirinae Techniques Testing Therapeutic Transgenic Organisms Transplantation Viral Vector Virus Vision cellular transduction effective therapy efficacy testing ganglion cell gene therapy human embryonic stem cell in vivo mouse model neuroprotection new therapeutic target novel overexpression promoter protective effect receptor retinal rods sigma-1 receptor stem cell therapy subretinal injection therapeutic target

项目摘要

DESCRIPTION (provided by applicant): Neurodegenerative eye diseases such as glaucoma cause blindness in a large population in the USA, yet effective treatments are lacking. The ultimate goal of this project is to develop new methods for rescue of retinal neurodegeneration by exploiting a unique endogenous neuroprotective agent, the ?1R chaperone, whose anti-apoptotic properties are being uncovered. Our in vivo studies suggest that the ? 1R is ROS (reactive oxygen species)-suppressing and is protective against retinal degeneration in ganglion cells as well as in photoreceptors. This project represents the first study to test the efficacy of the ? 1R-mediated retinal neuroprotection by ? 1R gene therapy, by overexpressing the ? 1R in stem cell-derived photoreceptors to enhance their post-transplantation survival, and by using the sigma-1 receptor knock model (? 1ko) combined with the chronic DBA/2J glaucoma model or rd10 model. In Specific Aim 1, we will examine the impact of the absence of the ? 1R for the viability of ganglion cells and photoreceptors in vivo in the DBA/2J model of glaucoma and the rd10 model of RP, respectively. Since we have discovered that ganglion cells of the ? 1ko mice are more susceptible to retinal degeneration than the wild type in an acute model, we will further test our finding using a new model crossed with the ? 1ko and the chronic glaucoma model of DBA/2J. We will also extend our investigation into a neuroprotective role of the ? 1R in retinal photoreceptors, which has yet to be defined, in a new model that we have generated by crossing ¿1ko and rd10. Retinal ROS levels will be compared in the presence and absence of the ? 1R in these models. In Specific Aim 2, we will test ? 1R gene therapy for rescue of retinal degeneration using lentiviruses. To increase the ? 1R abundance in the retina, we will transduce the rd10 mouse photoreceptors with lentiviruses harboring the ? 1R gene and the Opsin promoter using a technique of subretinal injection. Retinal functions of the transgenic eye overexpressing the ? 1R will be compared to that injected with the ? 1R-negative mock virus. In Specific Aim 3, we will explore the neuroprotective effect of the ? 1R on the post-transplantation survival of stem cell-derived photoreceptors. In stem cell therapy, elevated oxidative stress due to the death of rods, which are active major consumers of oxygen in the retina, imposes detrimental threat to the survival of transplanted photoreceptors. We will overexpress the ? 1R in human embryonic stem cell-derived photoreceptors to enhance their survival after transplantation into the rd10 retina. The pro-survival effect of the ? 1R on transplanted photoreceptors will be assessed in comparison to the control transplant cells that are transduced with the ? 1R-negative mock virus. Ultimately, our finding will lead to new ? 1R-targeted therapeutic strategies for rescue of the devastating retinal neurodegenerative diseases. PUBLIC HEALTH RELEVANCE: Neurodegenerative eye diseases such as glaucoma cause blindness in a large population in the USA. We will test gene therapy to rescue retinal neurodegeneration by exploiting a unique endogenous neuroprotective agent, the sigma-1 receptor chaperone.

描述(申请人提供)：神经退行性眼病，如青光眼，在美国导致大量人口失明，但缺乏有效的治疗方法。该项目的最终目标是通过开发一种独特的内源性神经保护剂-1R伴侣来开发拯救视网膜神经退行性变的新方法，其抗凋亡特性正在被发现。我们的体内研究表明，？1R是抑制ROS(活性氧)的，在神经节细胞和光感受器中对视网膜退行性变具有保护作用。这个项目代表了第一个测试药物疗效的研究通过？1R基因治疗、通过在干细胞来源的光感受器中过表达？1R以提高移植后存活率以及通过联合使用Sigma-1受体敲打模型(？1KO)和慢性DBA/2J青光眼模型或RD10模型来实现？1R介导的视网膜神经保护。在具体目标1中，我们将分别在DBA/2J青光眼模型和RP的RD10模型中检测？1R缺失对在体神经节细胞和光感受器活性的影响。由于我们已经发现？1KO小鼠的神经节细胞在急性模型中比野生型更容易发生视网膜变性，我们将使用与？1KO和DBA/2J慢性青光眼模型杂交的新模型进一步验证我们的发现。我们还将扩展我们的研究，在一个新的模型中，我们通过杂交1KO和rd10产生了尚未定义的视网膜光感受器中？1R的神经保护作用。在这些模型中，视网膜ROS水平将在存在和不存在？1R的情况下进行比较。在特定的目标2中，我们将测试？1R基因疗法，以挽救使用慢病毒的视网膜变性。为了增加视网膜中β1R的丰度，我们将利用视网膜下注射技术将携带β1R基因和Opsin启动子的慢病毒转导rd10小鼠光感受器。过量表达？1R的转基因眼的视网膜功能将与注射？1R阴性模拟病毒的转基因眼进行比较。在具体目标3中，我们将探讨？1R对干细胞来源的光感受器移植后存活的神经保护作用。在干细胞治疗中，由于视杆细胞的死亡而导致的氧化应激增加，视杆细胞是视网膜中活跃的氧气消耗者，对移植的光感受器的生存构成了有害的威胁。我们将在人类胚胎干细胞来源的光感受器中过表达？1R，以提高它们在移植到rd10视网膜后的存活率。？1R对移植的光感受器的促存活作用将与转导？1R阴性模拟病毒的对照移植细胞进行比较。最终，我们的发现将导致新的？1R靶向治疗策略，以拯救毁灭性的视网膜神经退行性疾病。公共卫生相关性：神经退行性眼病，如青光眼，在美国导致大量人口失明。我们将通过开发一种独特的内源性神经保护剂--Sigma-1受体伴侣来测试拯救视网膜神经退行性变的基因疗法。