Targeted Modification of Host and Proviral DNA to Treat Latent HIV Infection

描述(由申请人提供)：长期控制和治愈艾滋病毒的一个主要障碍是艾滋病毒在含有潜伏感染、静息和生产性感染的CD4T细胞的水库中持续存在。最近用抗HIV细胞的造血细胞移植(HCT)治愈了一例HIV患者，证明了这一策略的巨大潜力。不幸的是，对艾滋病毒具有抵抗力的捐赠者极其罕见，严重限制了这一方法。在这里，我们提出了一个多研究者计划，以研究患者和非人类灵长类动物中艾滋病毒储存库的生物学，并开发新的策略，用抗艾滋病毒的细胞重新填充造血系统。我们组建了一个由艾滋病毒、基因改造、转化研究和移植领域的领导者组成的团队。具体地说，我们将结合自体HCT和基因改造方法，使我们能够破坏自体干细胞中的CCR5受体，并破坏这些细胞中包含的整合的艾滋病毒前病毒。我们提出了5个高度集成的项目来追求我们的总体目标，并提出了5个核心来支持它们。项目1，造血细胞移植：清除艾滋病毒潜伏贮备库的平台(Ann Woolfrey，FHCRC)，将寻求澄清准备方案和清除艾滋病毒潜伏贮存库所需的供体移植物的相对贡献。项目2，ZFN修饰的干细胞根除艾滋病毒(菲利普·格雷戈里，Sangamo Biosciences)，将通过破坏CCR5基因的锌指核酸酶，寻求赋予移植接受者自身细胞对感染的稳定抵抗力。项目3，旨在控制非人类灵长类动物中艾滋病毒/希沃尔病毒的CCR5(Hans-Peter Kiem，FHCRC)，将使用艾滋病毒/希沃尔病毒感染的非人类灵长类动物模型，以确定CCR5基因座和Hct的基因改造是否可以提供对艾滋病毒的长期控制。项目4，通过工程归巢内切酶定向破坏整合的SHV，(Keith Jerome，FHCRC)将寻求通过设计高度特异性的归巢内切酶来直接解决病毒持久性的问题，该酶可以识别和突变受感染的猕猴造血细胞中的整合SHV前病毒。项目5，锌指核酸酶和归巢核酸内切酶的适体和树枝状分子递送(约翰·罗西，希望之城贝克曼研究所)将开发和优化递送策略，通过这种递送策略可以将ZFN和HES递送到他们的目标细胞。鉴于我们在移植和艾滋病毒研究方面的领先地位，我们相信我们有得天独厚的优势，可以将这些概念从高度相关的非人类灵长类HIV/SIV模型转移到HIV感染患者的治疗上。

DESCRIPTION (provided by applicant): A major obstacle to long-term control and cure of HIV has been the persistence of HIV in reservoirs that contain latently infected, resting, and productively infected CD4+ T cells. The recent cure of a patient with HIV by hematopoietic cell transplantation (HCT) with HIV-resistant cells has demonstrated the enormous potential of this strategy. Unfortunately, HIV-resistant donors are extremely rare, severely limiting this approach. Here we propose a multi-investigator program to study the biology of the HIV reservoir in patients and nonhuman primates, and to develop novel strategies to repopulate the hematopoietic system with HIV-resistant cells. We have assembled a team consisting of leaders in the fields of HIV, genetic modification, translational research, and transplantation. Specifically, we will combine autologous HCT with genetic modification approaches, allowing us to disrupt the CCR5 receptor in autologous stem cells, and also destroy the integrated HIV provirus contained within those cells. We propose 5 highly integrated projects in pursuit of our overall goal, and 5 cores for their support. Project 1, Hematopoietic Cell Transplant: Platform for Purging the Latent HIV Reservoir (Ann Woolfrey, FHCRC), will seek to clarify the relative contributions of the preparative regimen and the donor graft required for purging the latent HIV reservoir. Project 2, ZFN-Modified Stem Cells for HIV Eradication (Philip Gregory, Sangamo Biosciences), will seek to endow the transplant recipient's own cells with stable resistance to infection, via zinc finger nuclease disruption of the CCR5 locus. Project 3, CCR5 Targeting to Control HIV/SHIV in Nonhuman Primates (Hans-Peter Kiem, FHCRC), will use a non-human primate model of HIV/SHIV infection to determine whether genetic modification of the CCR5 locus and HCT can provide long-term control of HIV. Project 4, Targeted Disruption of Integrated SHIV by Engineered Homing Endonucleases, (Keith Jerome, FHCRC), will seek to directly address the problem of viral persistence, by designing highly specific homing endonucleases that can recognize and mutate integrated SHIV provirus in infected macaque hematopoietic cells. Project 5, Aptamer and Dendrimer Delivery of Zn Finger Nuclease and Homing Endonuclease mRNA and cDNA (John Rossi, Beckman Research Institute of the City of Hope), will develop and optimize delivery strategies by which ZFNs and HEs can be delivered to their target cells. Given our leading roles in Transplantation and HIV research, we believe we are uniquely positioned to move these concepts from a highly relevant nonhuman primate HIV/SHIV model toward a cure for HIV-infected patients.