HIV Cure with CCr5 (-) Human IPS Hematopoietic Stem Cells

描述：本提案的目的是开发一种有效的方法，为HIV感染者提供“功能性治愈”。该方法是基于这样的观察，即缺乏CCR5表达的受试者可以对HIV感染具有高度抗性。我们的假设是，造血CD34+干细胞和祖细胞（HSPC）可以通过CCR5基因的突变来抵抗HIV感染。这些细胞移植回自体供体将防止艾滋病毒复制和效果的“治愈”。“首先，来自未感染个体的纯化的人CD 34+细胞或外周血单核细胞将通过强调非整合载体的策略转化为诱导多能干细胞（iPS）。然后将这些iPS细胞进行遗传修饰，使其具有与细胞表面上不存在该受体表达相关的CCR 5的Δ 32 bp天然突变。值得注意的是，天然缺乏CCR 5表达的细胞被给予了几年后没有HIV感染证据的“柏林患者”。iPS衍生的CCR5突变细胞将转化为⑶ 34+细胞（即iPS衍生的⑶ 34 + HSPC），然后分化为造血子代细胞。这些细胞将在细胞培养中和移植到人源化小鼠中后评估对HIV感染的抗性和正常细胞功能。将对来自HIV感染者的基因修饰的CD 34+细胞进行相同的程序。这些研究旨在优化我们为HIV感染者提供iPS衍生的CD34+ HSPC的方法，以预防HIV疾病进展并可能建立“功能性治愈”。"

DESCRIPTION: 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 Δ32bp 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."