Retinal Gene Delivery by Adeno-Associated Virus

腺相关病毒的视网膜基因传递

• 批准号: 7473828
• 负责人: WILLIAM W HAUSWIRTH
• 金额: $ 42.35万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-08-01 至 2010-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7473828
• 关键词: Affect; Age; Animal Model; Apoptosis; Apoptotic; Behavior; Biological Preservation; Breeding; British Columbia; Capsid; Caspase Inhibitor; Cells; Childhood; Ciliary Neurotrophic Factor; Class; Classification; Clinical Trials; Collaborations; Complementary DNA; Data; Dependovirus; Disease; Effectiveness; Exhibits; Factor X; Gene Delivery; Gene Expression; Genes; Goals; Grant; Hand; Housing; Human; Individual; Knockout Mice; Link; Longevity; Malignant Neoplasms; Mediating; Modeling; Mus; Mutation; Opsin; Photoreceptors; Production; RPE65 protein; Range; Rattus; Replacement Therapy; Research Personnel; Resources; Retina; Retinal; Retinal Cone; Retinal Degeneration; Retinal Diseases; Retinal Dystrophy; Rodent Model; Safety; Serotyping; Stargardt' s disease; Structure; Testing; Therapeutic; Therapeutic Studies; Toxic effect; Transgenic Organisms; Translations; Vertebrate Photoreceptors; Washington; X-Linked Retinoschisis; adeno-associated viral vector; base; cell type; gene replacement; gene therapy; human BIRC4 protein; human disease; in vivo; interest; mouse model; mutant; neurotrophic factor; pre-clinical; promoter; research study; restoration; retinal rods; therapy duration; treatment duration; vector

DESCRIPTION (provided by applicant): We propose to continue using AAV vectors as a gene delivery vehicle to explore ways to rescue, restore or delay loss of retinal structure and function in animal models of retinal dystrophy (RD). We will focus primarily on seven recessive mouse models for human retinal degenerations representing a broad range of human RDs affecting rods, cones or the RPE. Four will be carried out primarily in-house while three are continuing collaborations with other investigators. Our overall hypothesis is that gene-based therapies will be clinically useful for a wide variety of RDs and that the safest and most effective AAV vectors will be those that incorporate serotypes and promoters with the ability to target passenger gene expression preferentially to the affected retinal cell type. We have chosen to focus on genetically and phenotypically well-characterized RD rodent models encompassing as wide a variety of disease classes and target cells as possible in order to fully assess the generality of our hypotheses. These include gene replacement therapy for the rd10 mouse, a rod PDEbeta recessive RP model, the ABCA4 knock out mouse, a model for recessive Stargardt's disease, the RS-1 knock out mouse, a model for X-linked Retinoschisis, the rd12 mouse, a model for RPE65 LCA, the LRAT knock out mouse, a model for recessive childhood RP, the Cpfl3 mouse, a model for Achromotopsia 2 (recessive GANT2 mutation), and the Cpfl1 mouse, a model for cone PDE6c recessive cone dystrophy. In addition we will test the efficacy of X-linked Inhibitor of Apoptosis (XIAP) in many of these mouse models by first crossing them into a transgenic line containing constitutively expressed XIAP in photoreceptors and RPE cells. For those models that respond, AAV-XIAP therapy will be tested. This proposal will also support vector production for more general retinal survival strategies through collaborations: AAV-XIAP, AAV-CNTF and/or AAV-CNTFalpha vectors will be employed in the rds mouse, the Cancer Associated Retinopathy (CAR) rat and in the P23H transgenic rat. In most cases we have already obtained initial proof-of-principle therapeutic results while for a few such experiments are just beginning. Our goals will be to characterize the magnitude and duration of therapy, to determine the age window within which therapy is effective and to define any retino-toxic effects of therapy, particularly over longer post-treatment periods. Ultimately we view this broad but systematic approach to validating the effectiveness and safety of AAV-vectored RD gene therapies as a necessary part of the initial preclinical safety/efficacy data required before translation to clinical trials on each analogous human disease could begin.

描述（由申请人提供）：我们建议继续使用AAV载体作为基因传递载体，探索在视网膜营养不良（RD）动物模型中挽救、恢复或延缓视网膜结构和功能丧失的方法。我们将主要关注人类视网膜变性的七种隐性小鼠模型，这些模型代表了影响视杆细胞、视锥细胞或视网膜色素上皮的广泛人类视网膜变性。其中四项将主要在内部进行，另外三项将继续与其他调查人员合作。我们的总体假设是，基于基因的疗法将在临床上适用于各种 RD，并且最安全和最有效的 AAV 载体将是那些整合了血清型和启动子的载体，能够优先将过客基因表达靶向受影响的视网膜细胞类型。我们选择关注遗传和表型特征良好的 RD 啮齿动物模型，涵盖尽可能广泛的疾病类别和靶细胞，以便充分评估我们假设的普遍性。其中包括 rd10 小鼠的基因替代疗法、杆状 PDEbeta 隐性 RP 模型、ABCA4 敲除小鼠、隐性 Stargardt 病模型、RS-1 敲除小鼠、X 连锁视网膜劈裂模型、rd12 小鼠、 RPE65 LCA 模型、LRAT 敲除小鼠、隐性儿童 RP 模型、Cpfl3 小鼠、全色盲 2 模型（隐性 GANT2 突变）和 Cpfl1 小鼠（视锥细胞 PDE6c 隐性视锥细胞营养不良模型）。此外，我们将首先将这些小鼠模型杂交到光感受器和 RPE 细胞中含有组成型表达 XIAP 的转基因系中，以测试 X 连锁细胞凋亡抑制剂 (XIAP) 在许多此类小鼠模型中的功效。对于那些有反应的模型，将测试 AAV-XIAP 疗法。该提案还将通过合作支持载体生产，以实现更通用的视网膜生存策略：AAV-XIAP、AAV-CNTF 和/或 AAV-CNTFalpha 载体将用于 rds 小鼠、癌症相关视网膜病变 (CAR) 大鼠和 P23H转基因大鼠。在大多数情况下，我们已经获得了初步的原理验证治疗结果，而少数此类实验才刚刚开始。我们的目标是描述治疗的强度和持续时间，确定治疗有效的年龄窗口，并确定治疗的任何视网膜毒性效应，特别是在较长的治疗后时期。最终，我们认为这种广泛但系统的方法来验证 AAV 载体 RD 基因疗法的有效性和安全性，是在开始转化为每种类似人类疾病的临床试验之前所需的初始临床前安全性/有效性数据的必要组成部分。