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: 9192555
• 负责人: Michael Massengill
• 金额: $ 4.15万
• 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/9192555
• 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; 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; abstracting; adeno-associated viral vector; 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; 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.

项目摘要/摘要 色素性视网膜炎（RP）是一组棘手的失明障碍，影响150万人 全世界。 RP的特征是刻板的光感受器死亡模式，杆凋亡 先于锥体的继发损失。重要的是，RP的生成疗法变得复杂 超过60个基因的突变会产生该疾病。但是，当前的证据表明一种共同的途径， 无论煽动突变如何，都会导致锥形死亡，从而代表潜在的治疗靶点。 尽管杆的保存将是理想的 驱动人类行为最重要方面的色觉。因此，这个长期目标 研究是为了揭示基因治疗方法的特定表征 增强所有形式的RP的锥生存力。目前的文献表明氧化应激，注射和 小胶质细胞的激活是由于鼠死亡的杆死亡而加剧了光感受器细胞损伤。 重要的是，在人RP患者眼中检测到了这些过程的间接证据。我的 中央假设预示着阻断氧化应激，注射和小胶质细胞激活的预示 保留RP中的锥生存力和视力。该假设将通过两个特定目的进行检验：1）执行 RAAV载体在小鼠模型中提供抗氧化剂或抗炎cDNA的临床前研究 常染色体显性RP（ADRP）。 2）在小鼠模型中表征小胶质细胞和巨噬细胞激活 ADRP。在AIM 1中，使用两种鼠标RP模型，我将测试两个raav矢量的效率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 （NRF2）或b）只需阻止两个炎症信号分子的活性，即NLRP3炎症体 和核因子Kappa B（NFκB）。在初步研究中阐述，AIM 2将表征不断发展的 视网膜中的M1（神经炎症）或M2（神经保护性）的小胶质细胞和巨噬细胞的表型 随着感光体在RP发病机理中死亡。此外，小胶质细胞的M1表型有助于其他 中枢神经系统（CNS）疾病，例如中风和阿尔茨海默氏病。那就更好地理解 特定目标1中使用的向量的受保护作用，它们阻止M1表型的能力将是 探索了体外和体内。在本应用程序中提出的研究很重要，因为它将 产生重要信息，以促进将锥体保存的成功疗法产生 受RP影响的人类患者。此外，这里测试的基因疗法方法是创新的，因为 它们显着偏离RP治疗的当前现状，涉及营养补充， 并且只需要在患者的一生中进行一次剂量。最后，由于视网膜是神经组织， 拟议的研究得出的结果可以扩展到其他CNS疾病，并具有氧化， 炎症或小胶质成分，例如脊髓损伤和帕金森氏病。