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基因突变对HIV感染产生抗性。这些细胞移植回 自体捐赠者将阻止艾滋病毒复制并实现“治愈”。“首先，纯化的人CD 34+细胞或 来自未感染个体的外周血单核细胞将被转化为诱导的多能干细胞。 (iPS)强调非整合载体的策略。这些iPS细胞将在基因上 修饰为具有与该受体表达缺失相关的CCR 5的<$32bp天然突变 在细胞表面。值得注意的是，将天然缺乏CCR 5表达的细胞给予“柏林患者”， 几年后没有感染艾滋病毒的证据。 iPS衍生的CCR 5突变细胞将转化为⑶ 34+细胞（即iPS衍生的⑶ 34 + HSPC）， 然后分化成造血子代细胞。这些细胞将被评估对艾滋病毒的抵抗力 在细胞培养中和移植到人源化小鼠中后，感染和正常细胞功能。相同的 将用来自HIV感染个体的遗传修饰的CD 34+细胞进行程序。 这些研究旨在优化我们为HIV-1提供iPS衍生的CD 34 + HSPC的方法。 感染者，以防止艾滋病毒疾病的进展，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 in the CCR5 gene. These cells transplanted back to autologous donors will prevent HIV replication and effect a "cure." First, purified human CD34+ cells or peripheral blood mononuclear cells from uninfected individuals will be converted into induced pluripotent stem (iPS) cells by strategies that emphasize non-integrating vectors. These iPS cells will then be genetically modified to have the ¿32bp natural mutation of CCR5 associated with the absence of this receptor expression on the cell surface. Notably, 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 converted into CD34+ cells (i.e. iPS-derived CD34+ HSPC) and then differentiated into hematopoietic progeny cells. These cells will be evaluated for resistance to HIV infection and normal cell function in cell culture and after transplantation into humanized mice. The same procedures will be undertaken with genetically modified CD34+ cells from HIV-infected individuals. These studies are directed at optimizing our approaches for providing iPS- derived CD34+ HSPC to HIV- infected individuals to prevent HIV disease progression and potentially establish a "functional cure."