Retinal-adhesive thermoresponsive gel for AAV-mediated gene delivery to the outer retina

用于将 AAV 介导的基因传递至外视网膜的视网膜粘附热敏凝胶

• 批准号: 10709508
• 负责人: William A. Beltran
• 金额: $ 71.98万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-30 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10709508
• 关键词: Address; Adhesives; Adverse effects; Affect; Alleles; Animal Model; Area; Biocompatible Materials; Canis familiaris; Cells; Clinic; Clinical; Clinical Trials; Complex; Data; Dependovirus; Development; Development Plans; Diffusion; Disease; Dose; Engineering; Ensure; Future; Gel; Gene Delivery; Genes; Human; Image; Immune response; Implant; Inflammation; Inflammatory; Injectable; Injections; Leber' s Hereditary Optic Neuropathy; Leber' s amaurosis; Libraries; Marketing; Mediating; Methods; Mutation; Nephronophthisis; Operative Surgical Procedures; Outcome; Outcome Study; Performance; Phenotype; Photoreceptors; Polymers; Procedures; Property; RPE65 protein; Research; Research Personnel; Retina; Retinal Degeneration; Retinal Diseases; Retinal Photoreceptors; Retinal gene therapy; Specificity; Structure of retinal pigment epithelium; Surface; System; Technology; Testing; Therapeutic; Time; Toxic effect; Toxicity Tests; Translations; Validation; Variant; Viral; Viral Vector; Viral load measurement; Virus Diseases; adeno-associated viral vector; biomaterial compatibility; canine model; cell type; ciliopathy; delivery vehicle; disease phenotype; dosage; effective therapy; expectation; fovea centralis; gene augmentation therapy; gene therapy; human model; implant compatibility; improved; in vivo; inherited retinal degeneration; innovation; intravitreal injection; macula; manufacture; member; neuroprotection; next generation; nonhuman primate; novel; novel strategies; optogenetics; particle; patient population; prevent; programs; single-cell RNA sequencing; skills; subretinal injection; success; transgene expression; vector; viscoelasticity

Project summary Until recently, there have been no effective treatments for retinal degenerations. FDA approval of Luxturna, the first gene therapy for bi-allelic mutations in RPE65, has opened the field for application to a broader range of retinal diseases. The rapidly growing number of clinical trials and emerging companies reflect the impact of this success and indicate the high expectations for retinal gene therapy. However, there is a significant need to develop new approaches for the many remaining forms of retinal degenerations, as RPE65-related dystrophies affect a very small population of patients. Moreover, the combination of the vector and surgical approach used for this disease is suboptimal for targeting the fovea in retinas with significant structural alterations. The major obstacles that must be addressed to improve clinical outcomes and extend the application of gene therapies to numerous retinopathies at various disease stages are: 1) efficient vector delivery to the central retina without damaging remaining photoreceptors, a significant, documented concern with sub-retinal injections in conditions where the retina is structurally compromised, 2) efficiently targeting gene delivery to affected cells, especially photoreceptors and RPE across the retina and 3) limiting the inflammatory/immune responses associated with intravitreal injections. These issues are relevant to all current and future retinal gene therapy programs. Here, we address each of these obstacles through development of an innovative new epiretinal gene therapy approach for NPHP5-LCA and RPE65-LCA2, in which a novel, biocompatible, retinal adhesive gel developed by our team releases high efficiency AAVs directly to the retina. We have created a comprehensive and efficient development plan that allows for rapid translation, drawing on the complementary skill sets of a team of experts with a track record of successful translational development. We will further develop the tunable, biocompatible gel and injection system to deliver these vectors directly to the retina, and we will incorporate a backing layer into the implant that allows for directionality of vector release for increased efficiency. We will determine the most efficient implant-compatible photoreceptor and RPE-targeting AAV vectors by utilizing our recently developed single cell RNA-Seq paradigm. We will fully validate this new gene therapy platform in two well-studied naturally occurring canine models of LCA that affect primarily the photoreceptors (NPHP5-LCA) and RPE (RPE65-LCA2), and we will characterize immune response and toxicity. The innovative approach developed herein will result in a new platform for direct, non-invasive and efficient AAV delivery to the retina, reducing diffusion and required dosage as well as the related immune response. This novel gene delivery platform has direct applicability to all outer retinal disease targets, paving the way forward for a safer, more efficient and targeted approach to treat a wide spectrum of disorders.

项目概要 直到最近，还没有针对视网膜变性的有效治疗方法。 FDA 批准 Luxturna 第一个针对 RPE65 双等位基因突变的基因疗法，为更广泛的应用开辟了领域 视网膜疾病。快速增长的临床试验数量和新兴公司反映了这一影响 成功并表明了对视网膜基因治疗的高度期望。然而，非常需要 开发新方法治疗许多剩余形式的视网膜变性，如 RPE65 相关营养不良 影响极少数患者。此外，所使用的载体和手术方法的结合 因为这种疾病对于针对具有显着结构改变的视网膜中央凹来说不是最佳选择。主要 为了改善临床结果和扩大基因疗法的应用必须解决的障碍 处于不同疾病阶段的许多视网膜病变是：1）有效地将载体递送至中央视网膜，而无需 损害剩余的光感受器，这是视网膜下注射的一个重要的、有记录的问题 当视网膜结构受损时，2) 有效地将基因递送至受影响的细胞，尤其是 视网膜上的光感受器和 RPE，以及 3) 限制与相关的炎症/免疫反应 玻璃体内注射。这些问题与所有当前和未来的视网膜基因治疗计划相关。这里， 我们通过开发创新的视网膜前基因治疗方法来解决这些障碍 适用于 NPHP5-LCA 和 RPE65-LCA2，其中我们团队开发了一种新型、生物相容性视网膜粘合凝胶 将高效 AAV 直接释放到视网膜。我们打造了全面高效的发展 允许快速翻译的计划，利用具有轨道的专家团队的互补技能 成功转化开发的记录。我们将进一步开发可调谐、生物相容性凝胶和 注射系统将这些载体直接传递到视网膜，我们将在载体中加入一个背衬层 植入物允许载体释放的方向性以提高效率。我们将确定最有效的 利用我们最近开发的单细胞，植入兼容的光感受器和 RPE 靶向 AAV 载体 RNA-Seq 范式。我们将在两个经过充分研究的自然发生的环境中充分验证这一新的基因治疗平台 主要影响光感受器 (NPHP5-LCA) 和 RPE (RPE65-LCA2) 的犬 LCA 模型，我们 将表征免疫反应和毒性。本文开发的创新方法将产生新的 用于直接、非侵入性且高效地将 AAV 递送至视网膜的平台，减少扩散和所需剂量 以及相关的免疫反应。这种新颖的基因传递平台可直接适用于所有外部 视网膜疾病目标，为更安全、更有效和更有针对性的方法治疗广泛的疾病铺平了道路 一系列疾病。