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: 10630097
• 负责人: Shannon Elizabeth Boye
• 金额: $ 67.36万
• 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/10630097
• 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; Rod; Role; Route; Safety; Sampling; Shapes; Site; Structure of retinal pigment epithelium; Testing; Thick; Thinness; Tissues; Toll-like receptors; Transgenes; Tropism; Variant; Vision; Visual Acuity; Work; X-Linked Retinoschisis; adeno-associated viral vector; clinical translation; disease-causing mutation; experimental study; fovea centralis; gene therapy; immune system function; improved; inclusion criteria; intravitreal injection; novel; promoter; rational design; 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“开发用于光感受器靶向的高效 AAV 载体”的竞争性更新 通过玻璃体”。请注意，由于初始资助期间取得的预付款以及我们改进的 了解该领域的挑战后，我们的标题已更改为“工程 AAV 以实现安全高效 基因递送到人类视网膜”以更广泛地涵盖我们的目标。 FDA 批准 Adeno 相关药物 基于病毒 (AAV) 的 RPE65-Leber 先天性黑蒙 (LCA2) 基因治疗巩固了基因治疗的地位 在当前的医疗实践中。然而，在一些患者的中央凹下注射载体导致视网膜中央 视力变薄和丧失。在无脉络膜血症中也观察到类似的视网膜厚度减少 临床试验。在更严重的情况下，例如 X 连锁视网膜劈裂症 (XLRS)，人们担心视网膜下 注射（SRI）会进一步损害患者的视网膜。由于大多数遗传性视网膜疾病 (IRD) 都是由 光感受器 (PR) 和视网膜色素上皮 (RPE) 特异性基因突变、基因发育 更安全、更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。瞄准中心凹 视锥细胞尤其重要，因为它们负责敏锐的日光视力。在初始资助期间， 我们开发了 AAV 衣壳，能够在灵长类动物玻璃体内注射 (IVI) 后进行有效的视网膜转导。 内界膜 (ILM) 是 AAV 通过此途径转导的主要障碍。然而，结果 来自使用 IVI AAV 的临床试验，显示剂量限制性炎症和 AAV 衣壳的中和作用 预先存在的抗体 (NAb) 表明宿主免疫系统是临床治疗更直接的“障碍” 翻译。眼睛的“免疫特权”可能导致人们对免疫系统作用的认识不足 塑造眼内递送 AAV 的结果。天然存在的衣壳抗体能够 通过玻璃体转导视网膜（即 AAV2）在高达 70% 的人类中普遍存在。因此，很大比例 的患者将不符合新兴疗法的纳入标准。在这里，我们提出基于强有力的实验 初步数据，以克服这些障碍。大部分工作将在灵长类动物（猕猴）中进行 这些屏障只能在具有眼部特征和免疫系统的动物的完整眼睛中重现 与人类相似。在目标 1 中，我们将通过以下方式增强玻璃体内递送 AAV 的转导和安全性： 改造衣壳和基因组以避免免疫识别。在目标 2 中，我们将增强视网膜转导 通过 subILM 递送 AAV，以实现内视网膜和外视网膜的高效和特异性转导。在目标 3 中，我们 将增强视网膜下递送的 AAV 的转导，这些 AAV 横向扩散到注射部位之外。向量 本提案中研究的方法和方法将对计划的临床试验产生直接影响，以解决 遗传性视网膜疾病以及AMD等非孤儿适应症。这些工具的开发 学术界（而不是工业界）将确保与更广泛的科学界共享资源的可用性 社区。

项目成果

Applying machine learning to predict viral assembly for adeno-associated virus capsid libraries.

• DOI: 10.1016/j.omtm.2020.11.017
• 发表时间: 2021-03-12
• 期刊: Molecular therapy. Methods & clinical development
• 作者: Marques AD;Kummer M;Kondratov O;Banerjee A;Moskalenko O;Zolotukhin S
• 通讯作者: Zolotukhin S

Night vision restored in days after decades of congenital blindness.

• DOI: 10.1016/j.isci.2022.105274
• 发表时间: 2022-10-21
• 期刊: ISCIENCE
• 影响因子: 5.8
• 作者: Jacobson, Samuel G;Cideciyan, Artur V;Ho, Allen C;Roman, Alejandro J;Wu, Vivian;Garafalo, Alexandra V;Sumaroka, Alexander;Krishnan, Arun K;Swider, Malgorzata;Mascio, Abraham A;Kay, Christine N;Yoon, Dan;Fujita, Kenji P;Boye, Sanford L;Peshenko, Igor V;Dizhoor, Alexander M;Boye, Shannon E
• 通讯作者: Boye, Shannon E

Identifying and Overcoming Challenges in Developing Effective Treatments for Usher 1B: A Workshop Report.

• DOI: 10.1167/tvst.12.2.2
• 发表时间: 2023-02-01
• 期刊: TRANSLATIONAL VISION SCIENCE & TECHNOLOGY
• 影响因子: 3
• 作者: Boye, Shannon E.;Durham, Todd;Laster, Amy;Gelfman, Claire M.;Sahel, Jose-Alai
• 通讯作者: Sahel, Jose-Alai

NF1 Is a Direct G Protein Effector Essential for Opioid Signaling to Ras in the Striatum.

• DOI: 10.1016/j.cub.2016.09.010
• 发表时间: 2016-11-21
• 期刊: CURRENT BIOLOGY
• 影响因子: 9.2
• 作者: Xie, Keqiang;Colgan, Lesley A.;Dao, Maria T.;Muntean, Brian S.;Sutton, Laurie P.;Orlandi, Cesare;Boye, Sanford L.;Boye, Shannon E.;Shih, Chien-Cheng;Li, Yuqing;Xu, Baoji;Smith, Roy G.;Yasuda, Ryohei;Martemyanov, Kirill A.
• 通讯作者: Martemyanov, Kirill A.

Capsid Mutated Adeno-Associated Virus Delivered to the Anterior Chamber Results in Efficient Transduction of Trabecular Meshwork in Mouse and Rat.

• DOI: 10.1371/journal.pone.0128759
• 发表时间: 2015
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Bogner B;Boye SL;Min SH;Peterson JJ;Ruan Q;Zhang Z;Reitsamer HA;Hauswirth WW;Boye SE
• 通讯作者: Boye SE