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

项目总结 直到最近，还没有有效的治疗视网膜退化的方法。FDA批准Luxturna，The 首次对RPE65双等位基因突变进行基因治疗，为更广泛的应用开辟了领域 视网膜疾病。迅速增长的临床试验和新兴公司的数量反映了这一影响 成功，表明人们对视网膜基因治疗寄予了很高的期望。然而，有一个重要的需求是 为RPE65相关性营养不良等多种残留形式的视网膜变性开发新的治疗方法 只会影响一小部分患者。此外，载体和手术入路的结合使用 因为这种疾病不适合于在结构发生显著变化的视网膜中靶向中心凹。少校 必须解决的障碍，以改善临床结果和扩大基因疗法的应用 在不同疾病阶段的许多视网膜病变是：1)有效的载体输送到中央视网膜，而不是 损害剩余的光感受器，这是视网膜下注射在有条件的情况下的一个重要的有记录的担忧 视网膜结构受损的地方，2)有效地将基因输送到受影响的细胞，特别是 视网膜上的光感受器和RPE以及3)限制与以下相关的炎症/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有关。这里, 我们通过开发一种创新的视网膜前基因治疗方法来解决这些障碍 对于NPHP5-LCA和RPE65-LCA2，其中我们团队开发了一种新型的、生物相容性的视网膜粘附胶 将高效的AAVs直接释放到视网膜。我们创造了全面高效的发展 允许快速翻译的计划，利用具有跟踪的专家团队的互补技能集 翻译开发的成功记录。我们将进一步开发可调的、生物相容的凝胶和 注射系统将这些载体直接输送到视网膜，我们将在 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 利用我们最新研制的单细胞植入相容的光感受器和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个新的基因治疗平台 犬LCA模型主要影响光感受器(NPHP5-LCA)和RPE(RPE65-LCA2)，而WE 将表征免疫反应和毒性。在此开发的创新方法将产生一种新的 将AAV直接、非侵入性和高效地输送到视网膜的平台，减少了扩散和所需剂量 以及相关的免疫反应。这种新型的基因传递平台对所有外源基因都有直接的适用性 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视网膜疾病铺平道路 疾病的谱系。

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.