A Phase I Study of Lentivirus Safety During T-Cell Immu

该提议的临床试验部分包括表达抗HIV siRNA的慢病毒载体转导的T细胞的I期研究（J. Zaia，P.I.）。这将是第一次在人类中进行抗HIV siRNA的试验，并将在City of Hope与C.六月在宾夕法尼亚大学和M。COH的詹森。这项研究的目的是确定慢病毒转导和扩增HAART失败的艾滋病患者T细胞的可行性和安全性。T细胞功能的临床前和临床方面将由C. 6月，转导和表达抗HIV RNA后。他将在扩增技术以及转导和选择后T细胞功能分析等领域为项目5的临床试验提供重要的GMP生产规模扩大支持。在T细胞选择系统的预期中，将探索IMPDH对T细胞功能的影响和慢病毒整合的安全性。具体目标如下： 目的1：确定RNAi-慢病毒转导的T细胞免疫疗法在AIDS背景下的可行性和安全性： 研究1：基于体外抗HIV效果的比较，选择慢病毒载体pHIV 7-shII用于临床I期研究（见初步结果）。该载体由J. Rossi和J.K. Yee，它是一种自我失活的慢病毒载体，编码靶向HIV-1 Rev中外显子的短发针RNAi。临床级T细胞产物将在输注研究患者之前和之后表征免疫功能、TCR库和对HIV的抗性。在本研究中，T细胞采集、转导和扩增将在核心C的BRICOH进行，受试者将在GCRC接受治疗。将评价输注前和输注后该转导对细胞的生物效应。 研究2：确定在本IPCP中开发的编码多重RNAi的慢病毒的安全性和相对有效性。使用目的1中评估的第一代pHIV 7-shlI慢病毒载体进行比较，第二项临床试验将评估用项目1-3中开发的多重RNAi载体转导的T细胞的相对存活率。在该第二临床试验中，受试者将接受用快速产生RNAi载体或用改进的载体转导的组合T细胞，然后观察在HIV存在下的相对T细胞存活。 目的2：表征体外扩增和选择后T细胞的功能： 将研究慢病毒RNAi转导后和用项目1-3中开发的新载体转导后扩增的T细胞的免疫功能。这项工作的一个特别的重点将是T细胞的选择和扩增的影响，使用IMPDH 2或MGMT为基础的方法，这些细胞在体外的免疫功能和抗HIV的效果。这将有助于研究1的安全性阶段和研究2的临床前阶段。

The clinical trial component of this proposal consists of a phase I study of lentiviral vector transduced T-cells expressing anti-HIV siRNA (J. Zaia, P.I.). This will be the first trial of anti-HIV siRNA in humans and will be performed at City of Hope with collaboration from C. June at UPENN and M. Jensen at COH. The goals of the study are to determine the feasibility and safety of lentiviral transduction and expansion of T-cells from AIDS patients who have failed HAART. The preclinical and the clinical aspects of T-cell function will be characterized by C. June after transduction and expression of anti-HIV RNAs. He will provide important GMP manufacturing scale up support for the clinical trial in Project 5 in the areas of expansion technology and analysis of the function of T-cell after transduction and selection. In anticipation of a T cell selection system, the effect of IMPDH on T cell function and the safety of lentivirus integration will be explored. The specific aims are as follows: Aim 1: To determine the feasibility and safety of RNAi-lentivirus transduced T cell immunotherapy in the setting of AIDS: Study 1: The lentivirus vector, pHIV7-shII, has been selected for clinical phase I studies based on comparative anti-HIV effect in vitro (see PRELIMINARY RESULTS). This vector was developed in our current IPCP activity by J. Rossi and J.K. Yee, and it is a self-inactivating lentivirus vector encoding a short hair-pin RNAi targeting an exon in HIV-1 rev. The clinical grade T cell product will be characterized for immune function, TCR repertoire, and resistance to HIV before and after infusion into research patients. In this study, the T cell collection, transduction, and expansion will be performed at BRICOH in Core C, and the subjects will be treated in the GCRC. The biologic effect of this transduction on cells pre- and post-infusion will be evaluated. Study 2: To determine the safety and relative efficacy of a lentivirus encoding a multiplex RNAi developed in this IPCP. Using the first generation pHIV7-shlI lentivirus vector evaluated in aim 1 for comparison, a second clinical trial will evaluate the relative survival of T cells transduced with a multi-plexed RNAi vector developed in Projects 1-3. In this second clinical trial, subjects will receive combined T cells transduced with either the fast generation RNAi vector or with the improved vector and then observed for relative T cell survival in the presence of HIV. Aim 2: To characterize the function of T cells after in vitro expansion and selection: The immunologic function of T cells expanded after lentivirus RNAi transduction and after transduction with new vectors developed in Projects 1-3 will be studied. A particular focus of this work will be the effect of T cell selection and expansion, using either IMPDH2- or MGMT-based methods, on the immunologic function of these cells in vitro and on the anti-HIV effect. This will contribute to the safety phase of study #1 and to the pre-clinical phase of study #2.