HIV cure with CCR5 (-) human IPS hematopoietic stem cells

摘要 该提案的目的是开发一种有效的方法，为艾滋病毒提供“功能性治疗”- 感染的人。该方法是基于观察到缺乏CCR 5表达的受试者可以被诱导。 对艾滋病毒感染有很强的抵抗力。我们假设造血CD 34+干细胞和祖细胞 （HSPC）可以通过CCR 5基因的突变或缺失对HIV感染产生抗性。这些细胞 移植回自体捐献者将阻止艾滋病毒复制并实现“治愈”。“首先，净化人类 来自未感染个体的CD 34+干细胞或外周血单核细胞将被转化为 通过各种策略诱导多能干（iPS）细胞，重点是非整合载体。 然后，这些iPS细胞将被遗传修饰为缺乏CCR 5基因或具有CCR 5基因的天然突变。 CCR 5。将天然缺乏CCR 5表达的细胞给予没有HIV证据的“柏林患者” 几年后感染。 iPS衍生的CCR 5突变细胞将分化成⑶ 34+细胞（即iPS衍生的HSPC），然后 分化成造血后代细胞。将评估这些细胞的正常细胞功能， 在细胞培养中和移植到人源化小鼠中后，抗HIV感染。相同的程序 将用来自HIV感染者的CD 34+细胞进行。 这些研究旨在优化我们向HIV感染个体提供iPS-HSPC的方法， 预防HIV疾病进展，并可能建立“功能性治愈”。"

Abstract The objective of this proposal is to develop an effective method for providing a "functional cure" for HIV- infected individuals. The approach is based on the observation that subjects lacking CCR5 expression can be highly resistant to HIV infection. Our hypothesis is that hematopoietic CD34+ stem and progenitor cells (HSPC) can be made resistant to HIV infection via mutation or deletion in the CCR5 gene. These cells transplanted back to autologous donors will prevent HIV replication and effect a "cure." First, purified human CD34+ stem cells or peripheral blood mononuclear cells from uninfected individuals will be converted into induced pluripotent stem (iPS) cells by a variety of strategies with emphasis given to non-integrating vectors. These iPS cells will then be genetically modified to either lack the CCR5 gene or have the natural mutation of CCR5. Cells naturally lacking CCR5 expression were given to the "Berlin patient" who has no evidence of HIV infection after several years. The iPS-derived CCR5-mutated cells will be differentiated into CD34+ cells (i.e. iPS-derived HSPC) and then differentiated into hematopoietic progeny cells. These cells will be evaluated for normal cell function and resistance to HIV infection in cell culture and after transplantation into humanized mice. The same procedures will be undertaken with CD34+ cells from HIV-infected individuals. These studies are directed at optimizing our approaches for providing iPS-HSPC to HIV-infected individuals to prevent HIV disease progression and potentially establish a "functional cure."