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

使用 CCr5 (-) 人 IPS 造血干细胞治愈 HIV

基本信息

批准号：
9052116
负责人：
JAY A LEVY
金额：
$ 69.77万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-05-01 至 2018-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9052116
关键词：
Address Alleles Anti-Retroviral Agents Autologous Back Berlin Binding Sites Blood Blood Cells CCR5 gene CD34 gene Cell Culture Techniques Cell Transplants Cell physiology Cell surface Cells Clinical Defect Disease Progression Gene Expression Gene Mutation Gene-Modified Genes Genetic Engineering Goals HIV HIV Infections HIV resistance HIV-1 Health Hematopoietic Hematopoietic System Hematopoietic stem cells Human Immune Immunocompromised Host Immunodeficient Mouse In Vitro Individual Laboratories Measures Methods Modification Mus Mutate Mutation Normal Cell Patients Peripheral Blood Mononuclear Cell Pharmaceutical Preparations Population Procedures Resistance Sendai virus Stem cells Surface Testing Tissues Transplantation base cellular engineering cellular transduction humanized mouse immune function in vivo induced pluripotent stem cell mouse model peripheral blood prevent receptor expression reconstitution research study stem transcription activator-like effector nucleases transcription factor vector viral detection

项目摘要

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."

描述：该提案的目的是开发一种为艾滋病毒感染者提供“功能治疗”的有效方法。该方法基于这样的观察结果，即缺乏CCR5表达的受试者可以高度抵抗HIV感染。我们的假设是，可以通过CCR5基因突变使造血CD34+茎和祖细胞（HSPC）具有抗HIV感染的能力。这些细胞移植回自体供体将防止HIV复制并影响“治愈”。首先，通过强调非整合载体的策略，将纯化的人类CD34+细胞或未感染个体的外周血单核细胞转化为诱导的多能干（IPS）细胞。然后，这些IPS细胞将通常被修饰，以使CCR5的Δ32bp自然突变与细胞表面上没有该受体表达相关的CCR5。值得注意的是，几年后没有艾滋病毒感染的“柏林患者”，自然缺乏CCR5表达的细胞。 IPS衍生的CCR5突变细胞将转换为CD34+细胞（即IPS来源的CD34+ HSPC），然后区分为造血后代细胞。将评估这些细胞在细胞培养中对HIV感染和正常细胞功能的抗性以及移植到人源化小鼠中的抗性。将使用来自HIV感染的个体的一般修饰的CD34+细胞进行相同的程序。这些研究旨在优化我们的方法，以向感染HIV感染的个体提供IPS衍生的CD34+ HSPC，以防止HIV疾病进展并有可能建立“功能治疗”。