HAIRPIN RIBOZYME GENE THERAPY FOR HCV INFECTION

描述：（改编自研究者摘要）本II期SBIR 该提案旨在开发发夹核酶（RZ）基因治疗， 丙型肝炎病毒（HCV）感染的治疗HCV是导致 非甲非乙型肝炎，这是一个严重的世界性健康问题， 核酶是RNA分子，其可以被工程化以切割和降解。 以序列特异性的方式与其他RNA分子结合。调查人员认为 HCV特别适合于核酶介导的治疗，因为病毒 基因组仅以RNA的形式存在，并且因为HCV仅编码 一个主要的成绩单。因此，申请人推断， 该RNA内靶序列应消除病毒表达。Immusol拥有 产生了两个靶向HCV正链RNA的发夹核酶， 在该项目的第一阶段，确定了其他Rz， 负链，由于其低表达而成为更有吸引力的靶 水平在本II期提案中，主要研究者提出 优化了前六位Rz的催化活性和稳定性。 肝细胞特异性启动子将用于将Rz表达为单个或多个启动子。 多顺反子转录物和这些多核酶载体的有效性 将在组织培养报告模型中进行测试。HCV的发展 复制系统在肝细胞培养提出，为最终的测试， 在大鼠毒性研究中，在临床前安全性评价之前使用核酶载体。 拟议商业应用：不可用

DESCRIPTION: (Adapted from the investigator's abstract) This Phase II SBIR proposal is aimed at the development of hairpin Ribozyme (RZ) gene therapy for the treatment of Hepatitis C virus (HCV) infection. HCV is the major cause of non-A, non-B hepatitis, which is a serious worldwide health problem, and ribozymes are RNA molecules that can be engineered to cleave and inactivate other RNA molecules in a sequence-specific fashion. The investigators believe that HCV is especially suited for ribozyme-mediated therapy, because the viral genome is present exclusively in the form of RNA and because HCV encodes only one primary transcript. Therefore, the applicants reason that cleavage of any target sequence within this RNA should abolish virus expression. Immusol has generated two hairpin ribozymes targeting the HCV positive strand RNA, and in the Phase I component of this project, additional Rz were identified targeting the negative strand, an even more attractive target due to its low expression level. In this Phase II proposal, the Principal Investigator proposes the optimization of the catalytic activity and stability of the top six Rz. Hepatocyte specific promoters will be used to express the Rz as single or polycistronic transcripts and the effectiveness of these multi-ribozyme vectors will be tested in a tissue culture reporter model. Development of an HCV replication system in liver cell culture is proposed, for final testing of the ribozyme vector before pre-clinical safety evaluation in a rat toxicity study. PROPOSED COMMERCIAL APPLICATION: NOT AVAILABLE