Leveraging Genetic Engineering Towards a Functional Cure of HIV Infection

项目3旨在确定移植的CD 34+造血干细胞的基因修饰是否 干细胞（HSC）抵抗HIV感染联合移植CD 8+基因修饰 T细胞可以协同作用，阻断ART治疗后的病毒反弹， 有效根除艾滋病毒宿主。具体而言，我们将利用项目2的专业知识， （新型HSC植入调节方案）和项目1和4（强大的基因编辑） 方法，包括CRISPR/Cas9）来有效地修饰和移植HSC和自体移植物。 HSC移植方法中的CD 8 + T细胞。我们将在体内测试这些概念， 最近开发的CD 34 + HSC衍生的人源化BLT小鼠模型重现了HIV 艾滋病毒感染和人体免疫力，以完成“防御和摧毁”的两个阶段 HIV治愈策略：i）通过敲除细胞内的 CD 34+造血干细胞（HSC）中的HIV共受体CCR 5;和ii）增强CD 34+造血干细胞（HSC）中的HIV共受体CCR 5的能力。 自体成熟的CD 8 + T细胞，以发现和破坏残留的HIV感染细胞， 操纵与效应子功能、粘膜和储库归巢以及靶向相关的基因， 细胞识别我们假设，限制病毒传播， 同时通过自体CD 8 + T细胞应答增强免疫清除， 有可能实现HIV感染的功能性治愈。

Project 3 seeks to determine whether gene modification of transplanted CD34+ hemotopoietic stem cells (HSCs) to resist HIV infection combined with gene modification of transplanted CD8+ T cells can function coordinately to block viral rebound following sessation of ART and effectively eradicate HIV reservoirs. Specifically, we will leverage the expertise of Project 2 (novel HSC engraftment conditioning regimens) and Projects 1 and 4 (powerful gene-editing approaches including CRISPR/Cas9) to efficiently modify and engraft HSCs and autologous CD8+ T cells within an HSC transplant approach. We will test these concepts in vivo utilizing the recently developed CD34+ HSC-derived humanized BLT mouse model that recapitulates HIV infection and human immunity to HIV to accomplish the two stages of a `Defend and Destroy” HIV cure strategy: i) defend transplanted cells against future HIV infection by knocking out the HIV co-receptor CCR5 in CD34+ hematopoietic stem cells (HSCs); and ii) enhance the capacity of autologous, mature CD8+ T cells to find and destroy residual HIV infected cells by manipulating genes associated with effector function, mucosal and reservoir homing, and target cell recognition. We hypothesize that the combined approach of limiting viral spread while simultaneously enhancing immune clearance by autologous CD8+ T cell responses has the potential to achieve a functional cure of HIV infection.