Engineering Novel AAV Vectors for Retinal Gene Therapy

用于视网膜基因治疗的新型 AAV 载体工程

• 批准号: 7268010
• 负责人: DAVID V SCHAFFER
• 金额: $ 17.93万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-08-01 至 2009-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7268010
• 关键词: Address; Affect; Age related macular degeneration; Animals; Capsid; Capsid Proteins; Cells; Cessation of life; Clinic; Dependovirus; Disease; Disease model; Engineering; Gene Delivery; Gene Expression; Glaucoma; Human; In Vitro; Infection; Injection of therapeutic agent; Invasive; Label; Medicine; Molecular; Molecular Biology; Muller' s cell; Neuroglia; Neurons; Numbers; Peptides; Personal Satisfaction; Photoreceptors; Population; Property; Retina; Retinal; Retinal Diseases; Retinal Ganglion Cells; Safety; Technology; Therapeutic; Translating; Variant; Virus; Work; adeno-associated viral vector; base; cell type; cellular targeting; cellular transduction; directed evolution; gene therapy; in vivo; mutant; novel; novel strategies; promoter; receptor binding; subretinal injection; vector; viral gene delivery; virology

DESCRIPTION (provided by applicant): Gene therapy has vast potential for treating and potentially curing a wide variety of disorders, in particular in a number of retinal diseases including glaucoma, age-related macular degeneration, and photoreceptor diseases. However, gene delivery technologies require significant improvements in cellular targeting, efficiency, and safety before promising findings in animal studies can be translated to the clinic. In particular, for retinal gene therapy it would be highly advantageous to transduce a single cell type that spans the entire retina for the delivery and secretion of a general neuroprotective factor throughout the retina to protect numerous populations of neurons that are affected by different retinal diseases, from retinal ganglion cells to photoreceptors. In addition, ideally this single cell type should be accessible from an intravitreal injection, as subretinal injections are more invasive and disruptive to the retina. Unfortunately, there is no vector capable of efficiently infecting the cell type that meets these needs, Muller glia. Vectors based on adeno-associated virus (AAV) have proven themselves to be highly promising in numerous retinal disease models, but they are also unfortunately incapable of Muller cell infection. We have developed novel directed evolution technology to generate new mutants of AAV with new properties, including altered receptor binding, and we propose to evolve variants capable of efficient Muller cell transduction. In parallel, the basic mechanisms of AAV transduction of Muller cells will be explored. Finally, results will be translated to photoreceptor disease model. The novel approaches developed in this work will have general impact for the molecular engineering of enhanced viral gene delivery vehicles for a number of retinal diseases.

描述（由申请人提供）：基因疗法在治疗和潜在治愈多种疾病方面具有巨大潜力，特别是在许多视网膜疾病中，包括青光眼、年龄相关性黄斑变性和光感受器疾病。然而，基因传递技术需要在细胞靶向、效率和安全性方面进行显着改进，然后才能将动物研究中的有希望的发现转化为临床。特别是，对于视网膜基因治疗来说，转导跨越整个视网膜的单一细胞类型以在整个视网膜中递送和分泌一般神经保护因子以保护受不同视网膜疾病影响的大量神经元是非常有利的。从视网膜神经节细胞到感光细胞。此外，理想情况下，这种单细胞类型应该可以通过玻璃体内注射获得，因为视网膜下注射对视网膜更具侵入性和破坏性。不幸的是，没有一种载体能够有效感染满足这些需求的细胞类型，即穆勒胶质细胞。基于腺相关病毒（AAV）的载体已被证明在众多视网膜疾病模型中非常有前途，但不幸的是它们无法感染穆勒细胞。我们开发了新颖的定向进化技术来生成具有新特性（包括改变的受体结合）的 AAV 新突变体，并且我们建议进化出能够有效进行 Muller 细胞转导的变体。同时，还将探索 AAV 转导 Muller 细胞的基本机制。最后，结果将转化为光感受器疾病模型。这项工作中开发的新方法将对针对多种视网膜疾病的增强病毒基因传递载体的分子工程产生普遍影响。

项目成果

Retinoschisin gene therapy in photoreceptors, Müller glia or all retinal cells in the Rs1h-/- mouse.

• DOI: 10.1038/gt.2014.31
• 发表时间: 2014-06
• 期刊: GENE THERAPY
• 影响因子: 5.1
• 作者: Byrne, L. C.;Ozturk, B. E.;Lee, T.;Fortuny, C.;Visel, M.;Dalkara, D.;Schaffer, D. V.;Flannery, J. G.
• 通讯作者: Flannery, J. G.