Prevention of Inherited Retinal Diseases by Therapeutic Rare Earth Nanoparticles

通过治疗性稀土纳米颗粒预防遗传性视网膜疾病

基本信息

批准号：
7895588
负责人：
JAMES Francis MCGINNIS
金额：
$ 36.32万
依托单位：
UNIVERSITY OF OKLAHOMA HLTH SCIENCES CTR
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-08-01 至 2012-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7895588
关键词：
3-nitrotyrosine Achievement Acrolein Acute Address Affect Age related macular degeneration Alzheimer&apos s Disease Antibodies Antioxidants Apoptosis Apoptotic Aspirin Biological Assay Biological Factors Biological Preservation Blindness Blood Vessels Broccoli - dietary Cessation of life Chronic Complementary DNA DNA Microarray Chip Data Defect Deoxyguanosine Development Diabetic Retinopathy Disease Down-Regulation Effectiveness Electroretinography Enzymes Event Exhibits Fluorescein Fluoresceins Fluorescence Genes Glaucoma Growth Heel Hematoxylin and Eosin Staining Method Histology Human Immunoblotting Inflammatory Inherited Injection of therapeutic agent Knockout Mice Light Macular degeneration Mammals Messenger RNA Methods Microarray Analysis Microscopy Modeling Mouse Strains Mus Mutant Strains Mice Mutation Nerve Degeneration Neural Retina Neurodegenerative Disorders Neurons Oxidative Stress Oxides Parkinson&apos s Dementia Pathway interactions Peroxides Phase Photoreceptors Prevention Proteins Publishing Rare Earth Metals Rattus Reactive Oxygen Species Retina Retinal Retinal Degeneration Retinal Diseases Retinitis Pigmentosa Reverse Transcriptase Polymerase Chain Reaction Severity of illness Signal Transduction Pathway Solutions Staining method Stains Structure of retinal pigment epithelium Sulforaphane Superoxides Techniques Testing Therapeutic Time Transgenes Transmission Electron Microscopy Tube Up-Regulation Usher Syndrome VLDL receptor Vascular Endothelial Growth Factors Vision Western Blotting aqueous catalase cerium oxide nanoparticle effective therapy human TXN protein human disease immunocytochemistry in vivo inherited retinal degeneration nanoparticle neuron apoptosis overexpression oxidative damage particle prevent programs retina outer nuclear layer retinal neuron

项目摘要

There are now almost 200 genes and chromosomal loci which have been identified as causing some form of inherited retinal degeneration. Irrespective of the primary mutation or cause, all of these diseases are thought to share a major common event. This is "oxidative stress" as a result of a chronic or acute rise in Reactive Oxygen Species (ROS). Antioxidants and over-expression of antioxidant enzymes have been shown to inhibit the progression of many retinal diseases. Inorganic cerium oxide nanoparticles (nanoceria) are antioxidants which mimic the activities of catalase and super oxide dismutase by catalytically scavenging ROS and have been shown to prevent peroxide induced blindness in rats. We think of the nanoceria as being analogous to an aspirin for blindness in that they won't cure the primary defect but will decrease the severity of the disease. We have hypothesized, that because ROS represent a node common to many inherited blinding diseases, they also represent an "Achilles' heel" which can be specifically targeted using ROS-scavenging nanoceria. As a component of that strategy, nanoceria will be used in this study to inhibit the inherited programmed death of photoreceptors in a mouse strain, tubby, which is a phenotypic model for Usher's Syndrome. Specific Aim 1 will test the hypothesis that the nanoceria particles will destroy ROS and, by decreasing oxidative damage in retinal neurons, will inhibit retinal degeneration induced by the tubby mutation. The hypothesis that nanoceria will act synergistically with sulphoraphane (Specific Aim 2) or with the overexpression of the human thioredoxin transgene (Specific Aim 3) to provide enhanced and prolonged protection of the photoreceptor cells in the tubby retina will also be tested. Our preliminary and published data support the effectiveness of each of these therapies when used alone in the tubby mouse. The mechanisms by which the nanoceria function in the tubby mouse alone, and in combination with sulphoraphane, or the Trx transgene, will be identified by the following methods. Superoxide radicals in the retina will be assessed using a hydroxyethidine assay whereas H20 2 will be assayed with 2',T-dichlorodihydro- fluorescein-diacetate. ROS-induced damage will be visualized with antibodies against products of ROS activity including acrolein, nitrotyrosine and 8-hydroxy-2-deoxy-guanosine. The effects of the nanoceria on neuroprotective pathways will be analyzed by Western blots, cDNA micro arrays, and Real Time-PCR. Quantitative histology, using bright field microscopy on hematoxylin and eosin stained retinal sections, will be used to evaluate the morphological preservation of photoreceptor cells. Retinal function will be determined by electroretinography. We currently have large colonies of both the tubby and the Trx-tubby mice and will be able to complete the specific aims within two years. The successful achievement of our objectives should be directly relevant to most forms of inherited blindness in mice. The nanoceria are expected to function in humans as they do in other mammals and therefore should preserve vision and prevent blindness in humans.

现在有近200个基因和染色体基因座，已被确定为引起某种形式的遗传性视网膜变性。无论主要突变或原因如何，所有这些疾病都被认为具有主要常见事件。这是反应性氧（ROS）的慢性或急性升高的“氧化应激”。抗氧化剂和抗氧化剂酶的过表达已被证明可以抑制许多视网膜疾病的进展。无机氧化葡萄纳米颗粒（纳米核）是抗氧化剂，通过催化清除ROS模仿过氧化氢盐和超级氧化物歧化酶的活性，并已证明可以防止大鼠过氧化物诱导的失明。我们认为纳米症类似于阿司匹林的失明，因为它们无法治愈主要缺陷，但会降低疾病的严重程度。我们假设，由于ROS代表了许多遗传性盲疾病共有的节点，因此它们也代表了一种“阿喀琉斯”脚跟，可以使用ROS扫除纳米果皮专门针对。作为该策略的一个组成部分，本研究将使用纳米症来抑制小鼠菌株中的光感受器的遗传程序死亡，这是Usher综合征的表型模型。具体的目标1将检验以下假设：纳米粒颗粒将破坏ROS，并通过减少视网膜神经元的氧化损伤，将抑制由Tubby突变引起的视网膜变性。纳米症将与硫二烷（特定目标2）或人硫氧还蛋白转基因的过表达（特定目标3）的假设（特定目标2）（特定的目标3）可提供增强和延长对塔比视网膜中光感受器细胞的保护。我们的初步和已发布的数据在单独使用在小鼠小鼠中时，支持每种疗法的有效性。纳米症单独在小鼠中功能以及与硫富或TRX转基因结合使用的机制将通过以下方法鉴定。将使用羟基乙胺测定法评估视网膜中的超氧化物自由基，而H20 2将用2'，T-Dichlorodihydro-- 荧光素二乙酸。 ROS诱导的损伤将使用针对ROS活性产物在内的丙烯醛，亚硝基苯胺和8-羟基-2-脱氧 - 瓜氨酸的抗体可视化。纳米症对神经保护途径的影响将通过蛋白质印迹，cDNA微阵列和实时PCR分析。使用苏木精和曙红染色的视网膜切片上的明亮场显微镜进行定量组织学用于评估感光细胞的形态保存。视网膜功能将由电视图确定。目前，我们拥有大型塔比比（Tubby）和TRX-tubby小鼠的群体，并且可以在两年内完成具体目标。我们目标的成功成就应与小鼠中大多数形式的遗传失明直接相关。预计纳米症将像其他哺乳动物一样在人类中起作用，因此应保留视力并防止人类失明。