Viral vector technology for cell type specific gene delivery

项目摘要 基因治疗是一种很有前途的治疗许多疾病的方法。为了使基因疗法越来越成功， 必须克服三个障碍：我们需要（1）安全的、（2）高效的和（3）细胞类型特异性的病毒载体。 腺相关病毒（AAV）已成为一种病毒载体，它在人体内安全， 将转基因转移到分裂和停滞的细胞，并能够驱动长期表达。不幸的是， 当基因递送至特定细胞（例如，癌症）是最重要的和异位的 在健康细胞或组织中的表达对患者的安全构成风险。 我们最近报道了一种新型可配置病毒基因递送技术的工作原型。这项技术 由一个衣壳组成，我们通过基因工程使其表达一个接头结构域， 以形成抗体-AAV复合物。AAV的向性被重定向到抗体的 同源受体，其在靶细胞类型上表达，但不在脱靶细胞群上表达。 在这里，我们将采取接下来的关键步骤，以这个原型为基础，扩大我们技术的影响。我们 将提高复合物-AAV形成效率和感染性（目的1），全面地绘制额外的 跨AAV血清型的工程能力确定新的衣壳工程策略，并使机器- 学习引导的AAV设计（目标2），并作为概念证明，确定AAV的靶特异性和传播 复合材料在体内（目标3）。 这项工作的成果将是一个有效的病毒载体平台技术，使用抗体靶向基因， 递送到合理鉴定的细胞类型。这项技术将从根本上实现新的基因治疗模式 从长远来看，它将为遗传性疾病和癌症带来新的治疗方法。

Project Summary Gene therapy is a promising treatment for many diseases. For gene therapy to become increasingly successful, three hurdles must be overcome: We need viral vectors that are (1) safe, (2) efficient, and (3) cell type specific. Adeno-associated virus (AAV) has emerged as a viral vector that is safe in humans, efficient at delivering transgenes to both dividing and arrested cells, and able to drive long-term expression. Unfortunately, the broad tropism of AAV is detrimental when gene delivery to specific cells (e.g., cancer) is paramount and ectopic expression in healthy cells or tissues poses a risk to the patient’s safety. We recently reported a working prototype of a novel configurable viral gene delivery technology. This technology consists of a capsid that we genetically engineer to express an adapter domain to which we covalently attach monoclonal antibodies to form antibody-AAV composites. AAV tropism is redirected toward the antibody’s cognate receptor, which is expressed on a targeted cell type, but not off-target cell populations. Here, we will take the next critical steps to build on this prototype and broaden the impact of our technology. We will improve composite-AAV formation efficiency and infectivity (Aim 1), comprehensively map additional engineerable capacity across AAV serotypes identify new capsid engineering strategies and enable machine- learning guided AAV design (Aim 2) and, as a proof of concept, determine target specificity and spread of AAV composites in vivo (Aim 3). The outcome of this work will be a validated viral vector platform technology that uses antibodies to target gene delivery to rationally identified cell types. This technology will enable fundamentally new gene therapy paradigms and, in the longer term, lead to new therapeutic approaches for inherited disorders and cancer.