Engineering T cells to Provide Durable Control of HIV-1 Replication

 描述（由申请人提供）：我们的长期目标是建立一个抗HIV的免疫系统，在没有HAART的情况下控制HIV-1复制。根据该领域先前的研究，很可能两者都增强了艾滋病毒特异性免疫力，以及CD 4+细胞的数量。 要实现这一目标，需要对艾滋病毒感染有抵抗力的T细胞。在与Sangamo Biosciences的合作中，我们最近进行了一项临床试验，通过锌指核酸酶破坏CCR 5辅助受体，注入T细胞使HIV抵抗。在没有HAART的情况下，这项研究中一个人的病毒载量下降到检测限以下，表明输注的HIV-1抗性CD 4 T细胞能够控制HIV-1复制。我们上一个IPCP的另一个好处是共同开发（Penn和Sangamo）一种名为C34-CXCR 4的强效抗病毒构建体。在本申请中，我们将确定该构建体的临床实用性，并通过比较我们在CCR 5 ZFN研究中的临床数据，我们希望获得重要的见解，了解体内保护CD 4 T细胞所需的关键因素，并更好地研究CD 4 T细胞应答的部分恢复如何控制HIV-1复制。传统上，在我们的学术/行业计划项目赠款中，一个一致的主题是在临床上同时测试一个概念，同时进行研究，作为下一个临床试验的基础和理由。这一应用的一个关键组成部分是延续这一传统。我们的建议的要素是：1）C34-CXCR 4肽修饰的CD 4 T细胞在HIV-1中的I期研究（项目1，Tebas [Penn]）：该项目将测试在HIV-1感染个体中输注表达C34-CXCR 4融合构建体的自体T细胞的安全性和可行性。2)C34修饰的辅助受体作为HIV-1进入的有效反式显性抑制剂（项目2，Holmes [Sangamo]）：该项目将利用Sangamo的锌指核酸酶技术将C34-CXCR 4靶向至原代人T细胞中的CXCR 4位点，限制遗传毒性和载体沉默的可能性。还将努力了解这种构建体如何有效地对抗所有HIV-1菌株，并在可能的情况下提高其活性。3)设计T细胞以功能性治愈HIV-1感染（项目3：Riley [Penn]）。该项目旨在设计HIV-1抗性，HIV-1特异性CD 4 T细胞，可以在没有HAART的情况下在HIV-1感染的人源化小鼠模型中长期控制HIV-1复制。4)编程长期持久的HIV-1特异性T细胞应答（项目4：Wherry [Penn]）。该项目旨在使用最先进的分析技术和一系列相关模型从分子上定义促进人类CD 4 T细胞持久性的途径，以探测使CD 4 T细胞能够长时间保持高水平活性的因素。该计划由2个核心支持：核心A是行政核心（PI，Riley）;核心B是测序和病毒进化核心（PI，Bushman）。此外，我们的计划利用现有的医学院，ITMAT和CFAR核心，以促进成本分摊，避免资源重复。

 DESCRIPTION (provided by applicant): Our long-term goal is to build an HIV-resistant immune system that controls HIV-1 replication in the absence of HAART. Based on prior studies in the field, it is likely that both enhanced HIV-specific immunity, as well as a population of CD4 T cells resistant to HIV infection will be required to achieve this goal. In collaboration with Sangamo Biosciences, we recently performed a clinical trial that infused T cells rendered HIV resistant by zinc finger nuclease disruption of the CCR5 coreceptor. The viral load of one of the individuals in this study dropped below the limit of detection in the absence of HAART, suggesting that infused HIV-1 resistant CD4 T cells are capable of controlling HIV-1 replication. Another benefit of our last IPCP was the co-development (Penn and Sangamo) of a potent antiviral construct called C34-CXCR4. In this application, we will determine the clinical utility o this construct, and by comparing our clinical data from the CCR5 ZFN study, we hope to gain important insight into the key factors required to protect CD4 T cells in vivo and to better study how partial restoration of the CD4 T cell response enables control of HIV-1 replication. Traditionally, throughout our academic/industry program project grants, a consistent theme has been to concurrently test one concept in the clinic while performing research that will serve as the basis and rationale for the next clinical trial. A key component of this application is the continuation of that tradition. The elements of our proposal are: 1) A Phase I Study of C34-CXCR4 Peptide-Modified CD4 T Cells in HIV-1 (Project 1, Tebas [Penn]): This project will test the safety and feasibility of infusing autologous T cells expressing a C34-CXCR4 fusion construct in HIV-1 infected individuals. 2) C34-modified Coreceptors as Potent Trans-dominant Inhibitors of HIV-1 Entry (Project 2, Holmes [Sangamo]): This project will utilize Sangamo's zinc finger nuclease technology to target C34-CXCR4 into the CXCR4 locus in primary human T cells, limiting the genotoxicity and the possibility of vector silencing. Efforts wll also be made to understand how this construct is so effective against all strains of HIV-1 and to improve its activity if possible. 3) Designing T cells to Functionally Cure HIV-1 infection (Project 3: Riley [Penn]). This project seeks to engineer HIV-1 resistant, HIV-1 specific CD4 T cells that can control HIV-1 replication long-term in the absence of HAART in a humanized mouse model of HIV-1 infection. 4) Programming Long-Term Durable HIV-1 Specific T cell Responses (Project 4: Wherry [Penn]). This project aims to molecularly define the pathways that promote durability in human CD4 T cells using state of the art profiling technology and an array of relevant models to probe factors that enable CD4 T cells to maintain high levels of activity for prolonged periods of time. The Program is supported by 2 Cores: Core A is the administrative Core (PI, Riley); Core B is the Sequencing and Viral Evolution Core (PI, Bushman). In addition, our Program takes advantage of existing School of Medicine, ITMAT, and CFAR Cores to promote cost sharing and avoid duplication of resources.