Combination gene editing for local and systemic HIV resistance

摘要 基因编辑有望提高抗HIV基因疗法的精确性和安全性， 改造造血干细胞和祖细胞（HSPC），以重建柏林患者的元素 疗方靶向核酸酶技术已经被用于利用NHEJ介导的DNA断裂修复， 从而破坏CCR 5基因，这模仿了他的治疗方法的一个方面。然而，将这种治疗扩展到非- 接受工程化自体HSPC的癌症患者可能需要包括其他方法。为了做到这一点， 我们正在开发基于同源定向修复（HDR）的DNA断裂修复的替代途径。这可以 用于将功能获得性突变引入细胞限制因子，或指导受控分泌 可溶性抗病毒因子，包括广泛进入抑制剂eCD 4-IG。此外，位点特异性插入在 CCR 5基因座上的eCD 4-IG将联合收割机HIV抗性与系统保护结合起来，从而提供了一种抗HIV的免疫系统。 组合抗HIV方法。在目前的提案中，我们的目标是继续提高安全性和有效性， HSPC中的基因编辑，并应用该技术提供局部和全身的HIV抗性。我们将 联合收割机将我们已建立的抗艾滋病毒方法与新方向相结合， 抗体，从而提供新的合成免疫能力。个人策略与组合 将在适当的人源化小鼠模型中评估这些方法，以评估它们对HIV感染的影响 和潜在的水库。

ABSTRACT Gene editing holds the promise of enhancing the precision and safety of anti-HIV gene therapies based on engineering hematopoietic stem and progenitor cells (HSPC), in order to recreate elements of the Berlin Patient cure. Targeted nuclease technology is already being used to exploit NHEJ-mediated repair of DNA breaks and thereby disrupt the CCR5 gene, which mimics one aspect of his cure. However, extending this treatment to non- cancer patients, receiving engineered autologous HSPC, will likely need to include other approaches. To do this, we are exploiting the alternate pathway of DNA break repair based on homology-directed repair (HDR). This can be used to introduce gain-of-function mutations into cellular restriction factors, or to direct the controlled secretion of soluble anti-viral factors, including the broad entry inhibitor eCD4-Ig. Moreover, the site-specific insertion at eCD4-Ig at the CCR5 locus would combine HIV-resistance with systemic protection and thereby provide a combinatorial anti-HIV approach. In the current proposal we aim to continue to improve the safety and efficacy of gene editing in HSPC, and to apply the technology to provide both local and systemic HIV resistance. We will combine our established anti-HIV approaches with a new direction to engineer production of broadly neutralizing antibodies, and thereby provide a novel synthetic immune capability. Individual strategies and combination approaches will be evaluated in appropriate humanized mouse models, to evaluate their impact on HIV infection and the latent reservoir.