Engineering AAV for safe and efficient gene delivery to the human retina

设计 AAV 以将基因安全有效地传递到人类视网膜

• 批准号: 10004652
• 负责人: Shannon Elizabeth Boye
• 金额: $ 69.47万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-06-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10004652
• 关键词: Academia; Acute; Address; Affect; Affinity; Animals; Antibodies; Area; Biodistribution; Blindness; Bulla; Bypass; Capsid; Cells; Characteristics; Choroideremia; Clinical Trials; Collection; Communities; Cone; DNA cassette; Data; Dependovirus; Development; Dose; Dose-Limiting; Engineering; Ensure; Eye; Funding; Gene Delivery; Genes; Genome; Goals; Heparan Sulfate Proteoglycan; Human; Immune; Immune response; Immune system; Industry; Inflammation; Inherited; Injections; Innate Immune Response; Inner Limiting Membrane; Leber' s amaurosis; Libraries; Macaca; Medical; Methods; Mus; Oligonucleotides; Outcome; Patients; Peripheral; Photoreceptors; Positioning Attribute; Primates; RPE65 protein; Resource Sharing; Retina; Retinal Detachment; Retinal Diseases; Retinal Photoreceptors; Rod; Role; Route; Safety; Sampling; Shapes; Site; Structure of retinal pigment epithelium; Testing; Thick; Thinness; Tissues; Toll-like receptors; Transgenes; Tropism; Variant; Vertebrate Photoreceptors; Vision; Visual Acuity; Work; X-Linked Retinoschisis; adeno-associated viral vector; base; clinical translation; design; disease-causing mutation; experimental study; fovea centralis; gene therapy; immune system function; improved; inclusion criteria; intravitreal injection; novel; promoter; response; screening; subretinal injection; tool development; transduction efficiency; transgene expression; vector

ABSTRACT This is a competitive renewal of R01EY024280, “Developing Efficient AAV Vectors for Photoreceptor Targeting via the Vitreous”. Note that, due to advances made during the initial funding period, and our improved understanding of challenges in the field, our title has been changed to “Engineering AAV for safe and efficient gene delivery to the human retina” to more broadly encompass our goals. FDA approval of an Adeno associated virus (AAV)- based gene therapy for RPE65-Leber congenital amaurosis (LCA2) solidified gene therapy’s place in current medical practice. However, injection of vector under the fovea of some patients led to central retinal thinning and loss of visual acuity. Similar decreases in retinal thickness were also observed in Choroideremia clinical trials. In more severe conditions, like X-linked Retinoschisis (XLRS), there is concern that subretinal injection (SRI) will further damage patient retinas. Since most inherited retinal diseases (IRDs) are caused by mutations in photoreceptor (PR)- and retinal pigment epithelial (RPE)- specific genes, development of gene therapies that more safely and efficiently target these cells remains a significant, unmet need. Targeting foveal cones is especially important, as they are responsible for acute, daylight vision. During the initial funding period, we developed AAV capsids capable of efficient retinal transduction following intravitreal injection (IVI) in primate. The inner limiting membrane (ILM) is the major barrier to AAV transduction via this this route. However, results from clinical trials utilizing IVI AAVs that show dose-limiting inflammation, and neutralization of the AAV capsid by pre-existing antibodies (NAbs) implicate the host immune system as a more immediate ‘barrier’ to clinical translation. The eye’s ‘immune-privilege’ has perhaps led to an under appreciation of the immune system’s role in shaping the outcome of intra-ocularly delivered AAVs. Naturally occurring antibodies to capsids capable of transducing retina via the vitreous (i.e. AAV2) are prevalent in up to 70% of humans. As such, a large percentage of patients will not meet inclusion criteria for emerging therapies. Here we propose experiments, based on strong preliminary data, to overcome these barriers. The majority of work will be performed in primates (macaque) as these barriers can only be recapitulated in intact eyes of animals with ocular characteristics and immune systems similar to humans. In Aim 1, we will enhance transduction and safety of intravitreally delivered AAVs by engineering the capsid and genome to avoid immune recognition. In Aim 2, we will enhance retinal transduction by subILM delivery of AAVs to enable efficient and specific transduction of inner and outer retina. In Aim 3, we will enhance transduction by subretinally delivered AAVs that spread laterally beyond the injection site. Vectors and methods investigated in this proposal will have an immediate impact on planned clinical trials to address inherited retinal diseases as well as non-orphan indications such as AMD. Development of these tools by academia (rather than industry) will ensure the availability of shared resources with the broader scientific community.

抽象的 这是R01EY024280的竞争更新，“开发有效的AAV向量来靶向感光器 通过玻璃体”。请注意，由于最初的资金期间的进步，我们的改进 了解该领域的挑战，我们的标题已更改为“工程AAV，以确保安全有效 基因传递到人类视网膜”，以更广泛地涵盖我们的目标。FDA认可了Adeno相关的 基于病毒（AAV）的基因疗法RPE65- leber先天性肌膜（LCA2）固化基因疗法的位置 在当前的医疗实践中。但是，在某些患者的中央凹下注射矢量导致中央视网膜 视力变薄和视力丧失。在脉络膜血症中也观察到残留厚度的类似下降 临床试验。在更严重的条件下，例如X连锁视网膜（XLR），关注视网膜下 注射（SRI）将进一步损害患者视网膜。由于大多数继承的残留疾病（IRD）是由 光感受器（PR）和视网膜色素上皮（RPE）的突变 - 特定基因，基因的发展 更安全有效地靶向这些细胞的疗法仍然是一个重要的，未满足的需求。靶向凹 圆锥体特别重要，因为它们负责急性，日光视野。在最初的资金期间， 我们开发了在灵长类动物玻璃体内注射（IVI）后能够有效的视网膜转导的AAV衣壳。 内部限制膜（ILM）是通过该路线转导的主要障碍。但是，结果 通过使用IVI AAV的临床试验，显示限制剂量炎症和中和AAV CAPSID 通过预先存在的抗体（NABS）将宿主免疫系统隐含为临床的更直接的“障碍” 翻译。眼睛的“免疫特你”可能导致对免疫系统的角色表示赞赏 在塑造眼内递送的AAV的结果时。自然发生的抗CAPSIDS能力的抗体 通过玻璃体（即AAV2）转导视网膜在多达70％的人类中普遍存在。因此，很大一部分 患者将不符合新兴疗法的纳入标准。在这里，我们提出了基于强的实验 初步数据，以克服这些障碍。大多数工作将在私人（猕猴）中进行 这些障碍只能在具有眼特征和免疫系统的动物的完整眼中概括 类似于人类。在AIM 1中，我们将增强玻璃体内的传播和安全性通过 工程带有衣壳和基因组，以避免免疫识别。在AIM 2中，我们将增强更多的转移 通过subilm递送AAV，以实现内部和外视网膜的有效和特定的转移。在AIM 3中，我们 将通过接下来的接下递送的AAV来增强翻译，该AAV横向扩散在注射部位以外。向量 该提案中研究的方法将对计划的临床试验产生直接影响，以解决 继承的残留疾病以及诸如AMD之类的非孔子指示。开发这些工具 学术界（而不是行业）将确保具有更广泛的科学的共享资源可用性 社区。