TRANSDUCTION OF HIV POSITIVE CD34+ PERIPHERAL BLOOD PROGENITOR CELLS

HIV 阳性 CD34 外周血祖细胞的转导

• 批准号: 6303646
• 负责人: John A Zaia
• 金额: $ 3.56万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-12-01 至 2000-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6303646
• 关键词: AIDS; AIDS therapy; Retroviridae; clinical research; clinical trial phase I; gene therapy; genetic transduction; hematopoietic stem cells; hematopoietic tissue transplantation; homologous transplantation; human immunodeficiency virus 1; human subject; human therapy evaluation; ribozymes; tissue /cell culture

This is a phase I study to evaluate the safety, engraftment, and relative survival of single infusions of genetically engineered autologous CD34+ hematopoietic progenitor cells from HIV+ individuals without prior cytoreductive treatment. CD34+ cells will be enriched from peripheral blood obtained by apheresis from asymptomatic HIV+ individuals following mobilization with G-CSF. Isolated CD34+ cells will be divided into two cultures and grown on autologous stromal cultures for three days in growth medum containing the cytokines IL-3, IL-6 and SCF. During this time one culture will be transduced with a retroviral vector containing the anti-HIV double-ribozyme gene, which is directed toward the HIV tat and rev regions (L-TR/TAT-new, also referred to as TR/TAT-Rz and identified as TR/TAT(V) in the Drug Master File, appendix A.1 for reference authorization letters) and the second culture will be transduced with a control vector (LN; identified as LN (V) in the DMF) that does not encode for a ribozyme. The two cultures of engineered cells will then be mixed and infused into the subject. Following infusion, the ratios of cells containing each vector will be determined by vector-specific PCR from peripheral blood samples obtained at monthly intervals and from marrow samples obtained at 6,12, and 24 months. A relative increase in the frequency of cells containing the anti-HIV-1 TR/TAT ribozyme gene, compared to cells with the neutral marker gene, would imply that a selective survival advantage has been conferred to the cells by the anti-HIV-1 ribozymes. The goal of this project is to determine whether autologous PBPC that are transduced with a retrovirus containing a gene encoding ribozymes targeted to the tat and rev genes of HIV-1, can safely engraft after reinfusion I HIV-1 infected persons. Follow-up studies will determine: 1) The safety and feasibility of administering a double ribozyme-encoding retroviral vector in HIV+ subject by a single dose infusion of ex vivo transduced autologous CD34+ peripheral blood progenitor cells (PBPC). 2) Whether the tranduced cells are capable of engrafting and giving rise to transduced mature hematopoietic cels in the periphery without prior cyto-reduction treatment of the subject. 3) The comparative survivial of ribozyme-vector transduced HPC progeny cells versus control vector marked cells in the periphery.

这是一项I期研究，旨在评估单次输注来自HIV+患者的基因工程自体CD34+造血祖细胞的安全性、植入和相对存活率，而不需要事先进行细胞减少治疗。在G-CSF动员后，从无症状的HIV+患者的外周血中提取CD34+细胞。将分离的CD34+细胞分为两组，在含有细胞因子IL-3、IL-6和SCF的生长基质中培养3天。在此期间，将用含有抗HIV双核酶基因的逆转录病毒载体转导一种培养物，该逆转录病毒载体针对HIVTAT和REV区(L-tr/tat-new，也被称为tr/tat-Rz，在药物总档案中被鉴定为tr/tat(V)，参考授权书的附录A.1)，第二种培养物将被转导到不编码核酶的对照载体(LN；在DMF中被鉴定为LN(V))。然后，将两种培养的工程细胞混合并注入受试者体内。输注后，包含每个载体的细胞比例将通过载体特异性聚合酶链式反应从每月一次的外周血液样本和6个月、12个月和24个月的骨髓样本中确定。与含有中性标记基因的细胞相比，含有抗HIV-1核酶基因的细胞的频率相对增加，这意味着抗HIV-1核酶赋予了细胞选择性生存优势。本项目的目的是确定将含有针对HIV-1 TAT和REV基因的核酶基因的逆转录病毒转导的自体PBPC在回输HIV-1感染者后是否可以安全地植入体内。后续研究将确定：1)通过单次输注体外转导的自体CD34+外周血祖细胞(PBPC)，将双核酶编码的逆转录病毒载体应用于HIV+受试者的安全性和可行性。2)未经细胞减数处理的情况下，转导细胞能否在外周移植并分化为成熟的造血细胞。3)核酶载体转导的HPC子代细胞与对照载体标记的细胞在外围的相对存活率。