EFFICACY, DELIVERY, AND EXPRESSION OF HIV PROTECTIVE GENES IN HIV+ CHILDREN

HIV 保护基因在 HIV 儿童中的功效、传递和表达

• 批准号: 5205770
• 负责人: Gary J Nabel
• 金额: --
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/5205770
• 关键词: AIDS therapy; HIV infections; SCID mouse; child (0-11); clinical trials; cooperative study; drug adverse effect; drug delivery systems; drug screening /evaluation; gene therapy; genetic transduction; hematopoietic stem cells; human subject; human therapy evaluation; messenger RNA; monocyte; plasmids; polymerase chain reaction; transfection /expression vector

The replication of HIV-1 is critically dependent upon the regulatory protein, Rev. Previous studies performed in this laboratory have demonstrated that M10, a transdominant mutant form of Rev which interferes with normal Rev function, prevents viral replication in human lymphocytes without affecting T cell function. We have exploited the therapeutic potential of Rev M10 for the treatment of acquired immunodeficiency syndrome (AIDS), and have developed vectors that express this gene product. Preliminary experiments using human lymphocytes indicate that expression of Rev M10 provides significant protection against HIV infection. A human clinical protocol was subsequently developed which is designed to determine whether Rev M10 will prolong the survival of T cells in HIV-infected individuals. This protocol was approved by the Recombinant DNA Advisory Committee of the NIH and will begin in the near future in adult volunteers. Despite the similarities between adult and pediatric AIDS, there are a number of differences between them which distinguishes their natural history and the course of these diseases. For this reason, we now propose to develop human clinical studies and preclinical models to implement gene therapy in pediatric populations. The first goal is to implement gene therapy in pediatric groups by delivering Rev M10 vectors into the T cells of these patients. We will utilize vectors developed in separate studies, which incorporate different cis-activating regulatory elements to increase the expression of this protein in CD4+ cells. The vectors will be tested on T cells stimulated with anti-CD3/IL2 or anti-CD3/CD28. The second aim for this project will be to develop and evaluate gene transfer in hematopoietic stem cells. These studies will involve the use of bone marrow and/or umbilical cord blood to be transplanted into different subsets of children who have an intact thymus that still retains the ability to reconstitute the immune system. Particle-mediated gene transfer will be developed along with the retroviral vector technology to deliver genes into hematopoietic stem cells. The third aim is to optimize gene expression and analyze the ability of the transduced cells to engraft and persist during development in vivo. Initial studies will be performed in animal models. These essential studies will facilitate the initiation of human gene therapy trials for AIDS in children and allow them to proceed with greater safety and efficacy. At the same time that these studies serve as a paradigm for other gene therapy approaches to the treatment of AIDS, they will also provide fundamental information on the pathogenesis of AIDS in children.

HIV-1的复制严重依赖于监管 蛋白质，Rev。之前在该实验室进行的研究已经 证明 M10，Rev 的一种反显性突变体形式， 干扰正常的 Rev 功能，阻止病毒在人体中复制 淋巴细胞，而不影响 T 细胞功能。 我们已经利用了 Rev M10 治疗获得性疾病的治疗潜力 免疫缺陷综合症（艾滋病），并开发了表达载体 这个基因产物。 使用人类淋巴细胞的初步实验 表明 Rev M10 的表达提供了显着的保护 对抗艾滋病毒感染。 随后制定了人类临床方案 开发的旨在确定 Rev M10 是否会延长 HIV 感染者中 T 细胞的存活。 该协议是 经 NIH 重组 DNA 咨询委员会批准，并将 在不久的将来开始在成年志愿者中进行。 尽管有相似之处 成人和儿童艾滋病之间存在许多差异 它们之间的区别在于它们的自然历史和进程 这些疾病。 为此，我们现在提出要发展人类 实施基因治疗的临床研究和临床前模型 儿科人群。 第一个目标是实施基因治疗 通过将 Rev M10 载体递送至儿科群体的 T 细胞中 患者。 我们将利用在单独研究中开发的载体， 纳入不同的顺式激活调节元件以增加 该蛋白在 CD4+ 细胞中的表达。 这些载体将被测试 用抗 CD3/IL2 或抗 CD3/CD28 刺激 T 细胞。 第二个目标 该项目将开发和评估基因转移 造血干细胞。 这些研究将涉及使用骨 骨髓和/或脐带血被移植到不同的 具有完整胸腺且仍保留着 重建免疫系统的能力。 粒子介导的基因 转移将与逆转录病毒载体技术一起发展 将基因传递到造血干细胞中。 第三个目标是 优化基因表达并分析转导细胞的能力 在体内发育过程中植入并持续存在。 初步研究将 在动物模型中进行。 这些重要的研究将有助于 启动针对儿童艾滋病的人类基因治疗试验 让他们能够以更高的安全性和有效性继续进行。 同时 这些研究可以作为其他基因治疗方法的范例 对于艾滋病的治疗，他们还将提供基本信息 儿童艾滋病的发病机制。