RIBOZYME GENE THERAPY FOR CD4 AND CD34 CELLS

CD4 和 CD34 细胞的核酶基因治疗

• 批准号: 6381270
• 负责人: Flossie Wong-Staal
• 金额: $ 27.44万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-09-30 至 2002-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6381270
• 关键词: AIDS; AIDS therapy; cell differentiation; cell proliferation; cytokine receptors; gene mutation; gene therapy; helper T lymphocyte; hematopoietic stem cells; human immunodeficiency virus 1; method development; polymerase chain reaction; ribozymes; tissue /cell culture

DESCRIPTION: Any therapeutic regimens against HIV must address the issue of viral resistance and escape. Gene therapy is an attractive alternative or adjunct to combination drug therapy because large number of antiviral genes can be devised that target viral as well as host cell sequences. We have been developing optimized ribozymes (Rz's) that recognize the HIV genome at multiple conserved sites, and more recently, Rz's that cleave the chemokine receptor, CCR5, which is a critical co-receptor for primary HIV infection. Individually, these genes have been shown to effectively inhibit HIV replication in T cells, and ongoing efforts involve assembly of several of these genes into a single retroviral vector for clinical evaluation. Here we propose to construct and validate the safety and efficacy of a polyribozyme vector for eventual clinical application. In parallel, we will design the next generation of polyRz vectors, which may include ones designed against possible resistant virus mutations and additional co-receptors. The specific aims of project 1 are: (1) To construct and test a murine retroviral vector expressing polyribozymes (4 anti-HIV and 1 anti-CCR5) for efficacy and safety at the preclinical level. (2) To identify resistant mutants to the U5, vif and pol Rz's by randomization of the target sequences in an infectious HIV-1 clone and passage in Rz expressing cells. If mutations are identified, ribozymes targeting them will be designed and tested. (3) To examine the effects of constitutive and inducible expression of ribozymes against CCR5, CCR3 and CXCR-4 on helper T cell functions of antigen-specific CD4 cell clones and on hematopoietic progenitor cells on their proliferation, differentiation and progeny cell function in vitro. Thus, our goals are to prepare and validate the first generation of polyRz vector as well as to design the next generation of polyRz vector which incorporate Rz's against potential resistant mutations and other co-receptors.

描述：任何针对艾滋病毒的治疗方案都必须解决 病毒抵抗和逃逸。基因疗法是一种有吸引力的替代选择 联合用药的辅助疗法，因为大量的抗病毒基因 可以设计出针对病毒和宿主细胞的序列。我们有 一直在开发能够识别HIV基因组的优化核酶(Rz‘s) 多个保守的位点，以及最近裂解趋化因子的Rz 受体CCR5，它是原发HIV感染的关键辅助受体。 单独来看，这些基因已经被证明能有效地抑制艾滋病毒 在T细胞中复制，并且正在进行的努力包括组装几个 将这些基因整合成单一的逆转录病毒载体，用于临床评估。这里 我们建议构建并验证一种 多聚核酶载体的最终临床应用。同时，我们将 设计下一代PolyRz向量，其中可能包括 针对可能的抗药性病毒突变而设计，以及其他 共同受体。项目1的具体目标是：(1)建设和 检测表达多聚核酶的小鼠逆转录病毒载体(4抗HIV和1 抗CCR5)用于临床前水平的有效性和安全性。(2)至 用随机化方法鉴定对U5、VIF和PolRz‘s的抗性突变体 HIV-1感染性克隆和传代中的靶序列 表达细胞。如果发现突变，靶向它们的核酶 将被设计和测试。(3)研究结构性和非结构性因素的影响 抗CCR5、CCR3和CXCR-4核酶在辅助性T细胞上的诱导表达 抗原特异性CD_4细胞克隆的细胞功能及其对造血的影响 祖细胞对其增殖、分化和子代细胞的影响 在体外发挥作用。因此，我们的目标是准备和验证第一个 PolyRz矢量的生成以及设计下一代的 融合Rz抗潜在耐药突变的PolyRz载体 和其他辅助受体。