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: 10453146
• 负责人: William A. Beltran
• 金额: $ 69.28万
• 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/10453146
• 关键词: Address; Adhesives; Adverse effects; Affect; Alleles; Animal Model; Area; Back; 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; Mediating; Methods; Mutation; 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; improved; in vivo; inherited retinal degeneration; innovation; intravitreal injection; macula; 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; Address; Adhesives; Adverse effects; Affect; Alleles; Animal Model; Area; Back; 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; Mediating; Methods; Mutation; 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; improved; in vivo; inherited retinal degeneration; innovation; intravitreal injection; macula; 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.

项目摘要 直到最近，还没有有效的视网膜变性治疗方法。 FDA批准Luxturna， RPE65中双行性突变的第一个基因疗法已开放该领域，以应用于更广泛的范围 视网膜疾病。临床试验和新兴公司的迅速增长反映了这一点的影响 成功并表明对视网膜基因治疗的期望很高。但是，很有必要 作为RPE65相关的营养不良，为许多剩余的视网膜变性形式开发新方法 影响很少的患者。此外，使用的矢量和手术方法的结合 因为该疾病是针对具有重大结构改变的视网膜中央凹的优势。专业 必须解决的障碍以改善临床结果并将基因疗法的应用扩展到 在各种疾病阶段，许多视网膜病变是：1）有效的矢量传递到中央视网膜，而没有 损害剩余的光感受器，这是在条件下进行下视网膜注射的重要关注 如果视网膜在结构上受到损害，则2）有效地将基因递送到受影响的细胞中，尤其是 光感受器和RPE跨越视网膜，3）限制与之相关的炎症/免疫反应 玻璃体内注射。这些问题与当前和未来的视网膜基因治疗计划有关。这里， 我们通过开发创新的新型前基因治疗方法来解决这些障碍 对于NPHP5-LCA和RPE65-LCA2，我们的团队开发了一种新颖的，生物相容性的视网膜粘合剂凝胶 将高效率AAV直接释放到视网膜上。我们创造了一个全面有效的发展 允许快速翻译的计划，利用具有轨道的专家团队的补充技能集 成功翻译发展的记录。我们将进一步开发可调节的，生物相容性的凝胶和 注射系统将这些向量直接传递到视网膜，我们将在 允许释放向量的植入物以提高效率。我们将确定最有效的 通过使用我们最近开发的单细胞 RNA-seq范式。我们将在两个自然发生的两个研究中充分验证这个新的基因疗法平台 LCA的犬类模型主要影响感光体（NPHP5-LCA）和RPE（RPE65-LCA2），我们 将表征免疫反应和毒性。本文开发的创新方法将导致新的 直接，非侵入性和高效的AAV向视网膜传递的平台，减少扩散和所需剂量 以及相关的免疫反应。这个新颖的基因输送平台直接适用于所有外部 视网膜疾病的目标，铺平了前进的方向，以更安全，更高效，更有针对性 疾病范围。