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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个基因和染色体位点已被确定为导致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有一个主要的共同事件。这是由于活性氧（ROS）的慢性或急性上升而导致的“氧化应激”。抗氧化剂和抗氧化酶的过度表达已被证明可以抑制许多视网膜疾病的进展。无机氧化铈纳米颗粒（nanoceria）是通过催化清除ROS来模拟过氧化氢酶和超氧化物歧化酶的活性的抗氧化剂，并且已经显示出防止过氧化氢诱导的大鼠失明。我们认为纳米氧化铈类似于失明的阿司匹林，因为它们不会治愈主要缺陷，但会降低疾病的严重程度。我们假设，因为ROS代表许多遗传性致盲疾病的共同节点，它们也代表了“阿喀琉斯之踵”，可以使用ROS清除纳米铈特异性靶向。作为该策略的一个组成部分，纳米铈将用于本研究中，以抑制小鼠品系tubby中光感受器的遗传性程序性死亡，tubby是Usher综合征的表型模型。具体目标1将测试纳米铈颗粒将破坏ROS并且通过减少视网膜神经元中的氧化损伤将抑制由tubby突变诱导的视网膜变性的假设。还将测试纳米氧化铈将与萝卜硫烷（特异性目标2）或与人硫氧还蛋白转基因的过表达（特异性目标3）协同作用以提供对管状视网膜中的感光细胞的增强和延长的保护的假设。我们的初步和发表的数据支持这些疗法中的每一种单独用于矮胖小鼠时的有效性。通过以下方法来鉴定纳米铈在单独的以及与萝卜硫素或Trx转基因组合的矮胖小鼠中起作用的机制。视网膜中的超氧化物自由基将使用羟乙啶测定法进行评估，而H2 O2将使用2 '，T-二氯二氢- 荧光素二乙酸酯。ROS诱导的损伤将用针对ROS活性产物（包括丙烯醛、硝基酪氨酸和8-羟基-2-脱氧-鸟苷）的抗体显现。将通过蛋白质印迹、cDNA微阵列和真实的时间PCR分析纳米铈对神经保护途径的影响。将使用苏木精和伊红染色视网膜切片上的明视野显微镜进行定量组织学检查，以评价感光细胞的形态学保存。视网膜功能将通过视网膜电图测定。我们目前拥有大规模的tubby和Trx tubby小鼠群体，并将能够在两年内完成特定目标。我们目标的成功实现应该与小鼠中大多数形式的遗传性失明直接相关。纳米氧化铈预计在人类中的功能与在其他哺乳动物中一样，因此应该可以保护人类的视力并防止失明。