Viral vector technology for cell type specific gene delivery

项目摘要 基因疗法是治疗许多疾病的一种很有前途的疗法。为了让基因治疗变得越来越成功， 必须克服三个障碍：我们需要(1)安全、(2)有效和(3)细胞类型特异的病毒载体。 腺相关病毒(AAV)是一种对人类安全、高效的病毒载体。 转基因既可用于分裂细胞，也可用于阻止细胞，并能够驱动长期表达。不幸的是，宽阔的 当特定细胞(如癌症)的基因传递是最重要的和异位的时，AAV的趋向性是有害的 在健康细胞或组织中的表达对患者的安全构成了风险。 我们最近报道了一种新型可配置病毒基因传递技术的工作原型。这项技术 由衣壳组成，我们通过基因工程来表达我们共价连接的适配域 单抗形成抗体-AAV复合体。AAV的趋向性被重定向到抗体的 同源受体，表达在靶细胞类型上，但不表达在非靶细胞群上。 在这里，我们将采取下一步关键步骤，在这一原型的基础上，扩大我们技术的影响。我们 将提高复合AAV的形成效率和传染性(目标1)，全面规划其他 跨AAV血清型的可工程能力识别新的衣壳工程战略，并使机器- 学习指导的AAV设计(目标2)，并作为概念验证，确定AAV的目标特异性和传播范围 体内复合材料(目标3)。 这项工作的结果将是一种经过验证的病毒载体平台技术，它使用抗体来靶向基因 传递给合理确定的细胞类型。这项技术将从根本上实现新的基因治疗范例 从长远来看，这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.