Development of Allele Independent Gene Therapy Strategies for Autosomal Dominant Retinitis Pigmentsa

常染色体显性遗传性色素性视网膜炎的等位基因独立基因治疗策略的开发

• 批准号: 9325296
• 负责人: Michael Massengill
• 金额: $ 4.2万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-16 至 2020-08-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9325296
• 关键词: Affect; Age; Alleles; Alzheimer' s Disease; Anti-Inflammatory Agents; Anti-inflammatory; Antioxidants; Apoptosis; Area; Attenuated; Behavior; Biological Preservation; Blindness; Bone Marrow; Cell Death; Cells; Central Nervous System Diseases; Cessation of life; Chronic; Color Visions; Complementary DNA; Cone; Dark Adaptation; Defect; Detection; Development; Disease; Dose; Environment; Event; Eye; Fostering; Gene Mutation; Gene Transduction Agent; Generations; Genes; Genetic; Goals; Homeostasis; Human; In Vitro; Individual; Infiltration; Inflammasome; Inflammation; Inflammatory; Inflammatory Response; Inherited; Injury; Interleukin-1 beta; Knock-out; Knowledge; Laboratories; Lead; Light; Literature; Macrophage Activation; Mediating; Mentors; Microglia; Mus; Mutation; Myxoma virus; NF-kappa B; Nerve Tissue; Neurodegenerative Disorders; Ophthalmology; Outcome; Oxidative Stress; Parkinson Disease; Pathogenesis; Pathway interactions; Patients; Pattern; Peptides; Peripheral; Phenotype; Photoreceptors; Process; Proteins; Quality of life; Recombinant adeno-associated virus (rAAV); Research; Research Personnel; Retina; Retinal; Retinal Cone; Retinal Dystrophy; Retinitis; Retinitis Pigmentosa; Role; Signal Transduction; Signaling Molecule; Spinal cord injury; Stroke; TNF gene; Testing; Therapeutic; Viral Proteins; Vision; Work; Writing; attenuation; base; career; cell injury; cytokine; design; disease natural history; efficacy testing; experience; gene therapy; in vivo; innovation; macrophage; monocyte; mouse model; neuroinflammation; nuclear factor-erythroid 2; nutritional supplementation; photoreceptor degeneration; preclinical study; protective effect; public health relevance; response; retinal rods; skills; therapeutic target; transcription factor; vector

Project Summary/Abstract Retinitis pigmentosa (RP) is a group of intractable blindness disorders that affects 1.5 million individuals worldwide. RP is characterized by a stereotypical pattern of photoreceptor death whereby rod apoptosis precedes a secondary loss of cones. Importantly, generation of therapies for RP is complicated by the fact that mutations in over 60 genes can yield the disease. However, current evidence suggests that a common pathway, regardless of the inciting mutation, leads to cone death and thus represents a potential therapeutic target. Though preservation of rods would be ideal, it is the detection of light by cones that produces the high acuity, color vision that drives the most important aspects of human behavior. Therefore, the long-term goal of this research is to characterize the cone death pathway in order to uncover gene therapy approaches to specifically enhance cone viability in all forms of RP. The current literature suggests that oxidative stress, inflammation and activation of microglia occur as a result of rod death and exacerbate photoreceptor cell injury in murine RP. Importantly, indirect evidence of these processes has been detected in the eyes of human RP patients. My central hypothesis portends that blockade of oxidative stress, inflammation, and microglial activation will preserve cone viability and vision in RP. This hypothesis will be tested through two specific aims: 1) Perform preclinical studies of rAAV vectors delivering either antioxidant or anti-inflammatory cDNAs in mouse models of autosomal dominant RP (adRP). 2) Characterize microglial and macrophage activation in mouse models of adRP. In aim 1, utilizing two mouse models of RP, I will test the efficacy of two rAAV vectors that either a) enhance the signaling of an antioxidant transcription factor known as nuclear factor erythroid 2-related factor 2 (Nrf2), or b) simultaneously block the activity of two inflammatory signaling molecules, the Nlrp3 inflammasome and Nuclear Factor kappa B (NFκB). Expounding on preliminary studies, aim 2 will characterize the evolving phenotype of microglia and macrophage, either M1 (neuroinflammatory) or M2 (neuroprotective), in the retina as photoreceptors die during RP pathogenesis. Additionally, the M1 phenotype of microglia contributes to other central nervous system (CNS) disorders, such as stroke and Alzheimer’s disease. Thus, to better understand the protective effects of the vectors employed in specific aim 1, their ability to block the M1 phenotype will be explored both in vitro and in vivo. The research proposed in this application is significant because it would produce important information that will facilitate the generation of successful therapies that preserve cones in human patients affected by RP. Furthermore, the gene therapy approaches tested here are innovative because they deviate significantly from the current status quo of RP treatment, which involves nutritional supplementation, and would only require a single dose for the lifetime of the patient. Finally, since the retina is nervous tissue, results derived from the proposed research could be extended to other CNS diseases with an oxidative, inflammatory, or microglial component, such as seen in spinal cord injury and Parkinson’s disease.

项目总结/摘要 视网膜色素变性（RP）是一组难治性失明疾病，影响150万人 国际吧RP的特征在于感光细胞死亡的刻板模式， 会导致视锥细胞的二次丧失重要的是，RP的治疗方法的产生由于以下事实而复杂化： 超过60个基因的突变会导致这种疾病。然而，目前的证据表明，一种常见的途径， 不管诱发突变如何，其导致视锥细胞死亡，因此代表了潜在的治疗靶点。 虽然保存视杆细胞是理想的，但正是视锥细胞对光的探测产生了高的敏锐度， 色觉驱动着人类行为的最重要方面。因此，这一长期目标 研究的目的是描述视锥细胞死亡途径，以揭示基因治疗方法， 增强所有形式RP的视锥细胞活力。目前的文献表明，氧化应激，炎症和 小胶质细胞的激活是视杆细胞死亡的结果，并加剧了鼠RP中的感光细胞损伤。 重要的是，这些过程的间接证据已在人类RP患者的眼睛中检测到。我 中心假设预示着阻断氧化应激、炎症和小胶质细胞活化将 保留RP中的视锥细胞活力和视力。这一假设将通过两个具体目标进行检验：1）执行 rAAV载体递送抗氧化剂或抗炎cDNA在小鼠模型中的临床前研究 常染色体显性遗传RP（adRP）。2)表征小鼠模型中的小胶质细胞和巨噬细胞活化， adRP。在目的1中，利用两种RP小鼠模型，我将测试两种rAAV载体的功效，所述两种rAAV载体或a） 增强称为核因子红细胞2相关因子2的抗氧化转录因子的信号传导 （Nrf 2），或B）同时阻断两种炎性信号分子，Nlrp 3炎性体的活性 和核因子κ B（NFκB）。在阐述初步研究的基础上，目标2将描述 视网膜中小胶质细胞和巨噬细胞的表型，M1（神经炎症）或M2（神经保护） 因为光感受器在RP发病过程中死亡。此外，小胶质细胞的M1表型有助于其他 中枢神经系统（CNS）病症，如中风和阿尔茨海默病。为了更好地理解 在特定目的1中使用的载体的保护作用，其阻断M1表型的能力将被 在体外和体内都进行了探索。本申请中提出的研究是重要的，因为它将 产生重要的信息，这将有助于产生成功的治疗， 受RP影响的人类患者。此外，这里测试的基因治疗方法是创新的，因为 它们显着偏离了RP治疗的当前现状，其中涉及营养补充， 并且在患者的一生中仅需要单次剂量。最后，由于视网膜是神经组织， 从所提出的研究中得到的结果可以扩展到其他具有氧化， 炎性或小胶质细胞成分，如在脊髓损伤和帕金森病中所见。