RIBOZYME GENE THERAPY FOR CD4 AND CD34 CELLS

CD4 和 CD34 细胞的核酶基因治疗

• 批准号: 6177741
• 负责人: Flossie Wong-Staal
• 金额: $ 26.7万
• 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 2003-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6177741
• 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.

任何针对HIV的治疗方案都必须解决以下问题： 病毒抵抗和逃逸 基因治疗是一种有吸引力的替代方案， 辅助联合药物治疗，因为大量的抗病毒基因 可以设计成靶向病毒以及宿主细胞序列。 我们有 一直在开发最优化的核酶（Rz's）， 多个保守位点，最近还有切割趋化因子的Rz 受体，CCR 5，其是原发性HIV感染的关键共受体。 单独来看，这些基因已被证明可以有效抑制艾滋病毒 在T细胞中复制，并且正在进行的努力涉及组装几种 将这些基因整合到单个逆转录病毒载体中用于临床评估。 这里 我们建议构建并验证一种 多核酶载体，用于最终的临床应用。 同时，我们将 设计下一代polyRz载体，其中可能包括 针对可能的耐药病毒突变和其他 共受体 项目1的具体目标是：（1）建设和 测试表达多核酶（4种抗HIV和1种 抗CCR 5）在临床前水平的功效和安全性。 (2)到 通过以下随机化鉴定对U 5、vif和pol Rz的抗性突变体： 感染性HIV-1克隆中的靶序列和在Rz中的传代 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)为了检查构成和 辅助性T细胞上抗CCR 5、CCR 3和CXCR-4核酶诱导表达 抗原特异性CD 4细胞克隆的细胞功能和造血功能 祖细胞对其增殖、分化和子代细胞的影响 体外功能 因此，我们的目标是准备和验证第一个 产生polyRz载体以及设计下一代 掺入针对潜在抗性突变的Rz的polyRz载体 和其他辅助受体。