Develop humanized AAV vectors for liver targeting and neutralizing antibody evasion

开发用于肝脏靶向和中和抗体逃避的人源化 AAV 载体

基本信息

批准号：
10079155
负责人：
Chengwen Li
金额：
$ 24.93万
依托单位：
BEDROCK THERAPEUTICS, INC.
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-08-15 至 2022-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10079155
关键词：
Affect American Anatomy Animal Model Animals Antibody titer measurement Binding Blindness Blood Coagulation Factor Blood Component Removal Canis familiaris Capsid Cell Line Cells Clinic Clinical Clinical Research Clinical Trials DNA Shuffling Data Dependovirus Disease Dose Engineering Evaluation FDA approved Gene Delivery Gene Transfer Genetic Diseases Goals Hemophilia A Hepatocyte High Prevalence Human In Vitro Infection Injections Intravenous Immunoglobulins Libraries Liver Liver diseases Masks Mediating Mendelian disorder Mental Depression Methods Modeling Mus Muscular Atrophy Mutation Organ Patients Pharmaceutical Preparations Pharmacologic Substance Phase Phenotype Plasma Population Primates Property Quality of life Rare Diseases Safety Serotyping Spinal Surface System Technology Testing Therapeutic Therapeutic Effect Time Tissues Traction Translations Tropism Validation Variant Xenograft Model Xenograft procedure adeno-associated viral vector animal tissue base clinical development cost experimental study gene product gene therapy humanized mouse in vivo in vivo evaluation innovation mortality mouse model mutant neutralizing antibody novel pre-clinical safety study success transduction efficiency transgene delivery vector virtual

项目摘要

Adeno-associated virus (AAV) vectors have been successfully applied in clinical trials in patients with hemophilia. Two AAV based gene therapy drugs have been recently approved by the FDA. Luxturna has been valued at $850,000 for a one-time treatment for a rare form of blindness and Zolgensma at $2,100,000 for spinal muscle atrophy. Gene therapy with AAV vectors has shown a potentially huge market. Although successful in clinical studies, two concerns restrict a broader AAV vector application for patients with hemophilia who need systemic administration of AAV vector for effective liver targeting: low human hepatocyte transduction and neutralizing antibody (Nab)-mediated blocking of AAV transduction. Several approaches have been explored for AAV transduction enhancement or neutralizing antibody evasion. Engineering of the AAV capsid represents a very powerful and popular technology, and has been extensively studied to develop novel AAV vectors for transduction enhancement in pre-clinical animal models or Nab escape in vitro. It has been demonstrated that the results from mouse experiments do not recapitulate those of large animals such as primates and dogs, the data for AAV variants generated in animal cells and organs may not translate into human application. Recently, a mouse model xenografted with human hepatocytes has been used to develop AAV vectors for targeting gene therapy relative to human liver. We have isolated several AAV mutants from the liver of chimeric mice xenografted with human hepatocytes in the presence of human neutralizing antibodies (IVIG) using AAV shuffling library approach. In this application, we will study the ability of these mutants to transduce human hepatocytes and evade Nabs. Following the identification of the best mutants with efficient human hepatocyte transduction and high ability for Nab evasion, we will study the safety and long-term efficacy of these mutants for hemophilia gene therapy in a mouse model with AAV Nabs after a single injection. Successful validation of these mutants in hemophilic mice will provide a proof-of-concept for translation to larger models of hemophilia in hopes of safely and effectively treating hemophilia patients with AAV Nabs using a single dose.

腺相关病毒（AAV）载体已成功应用于糖尿病患者的临床试验血友病。两种基于 AAV 的基因治疗药物最近已获得 FDA 批准。卢克斯图纳已经用于治疗一种罕见失明的一次性治疗的价值为 850,000 美元，Zolgensma 的治疗价值为 2,100,000 美元脊髓肌肉萎缩。 AAV载体的基因治疗已显示出潜在的巨大市场。虽然尽管临床研究取得了成功，但有两个问题限制了 AAV 载体在患有以下疾病的患者中更广泛的应用：需要全身施用 AAV 载体以实现有效肝脏靶向的血友病：人肝细胞水平低转导和中和抗体 (Nab) 介导的 AAV 转导阻断。几种方法有已探索 AAV 转导增强或中和抗体逃避。 AAV 的工程设计衣壳代表了一种非常强大和流行的技术，并且已经被广泛研究以开发新颖的用于临床前动物模型转导增强或 Nab 体外逃逸的 AAV 载体。它一直证明小鼠实验的结果并不能概括大型动物的结果，例如对于灵长类动物和狗，动物细胞和器官中产生的 AAV 变体的数据可能无法转化为人类应用。最近，用人类肝细胞异种移植的小鼠模型已被用于开发用于针对人类肝脏的基因治疗的 AAV 载体。我们已经从 AAV 中分离出几个突变体在人中和抗体存在的情况下，用人肝细胞异种移植的嵌合小鼠的肝脏 (IVIG) 使用 AAV 改组库方法。在此应用中，我们将研究这些突变体的能力转导人肝细胞并逃避 Nabs。在通过有效的方法鉴定出最佳突变体后人肝细胞转导和Nab逃避能力强，我们将研究安全性和长期疗效这些突变体在单次注射 AAV Nabs 的小鼠模型中进行血友病基因治疗。在血友病小鼠中成功验证这些突变体将为翻译提供概念验证更大的血友病模型，希望能够安全有效地使用 AAV Nab 治疗血友病患者使用单剂量。