EVALUATION OF INHIBITION OF SHIV REPLICATION BY SIRNA VECTORS

SIRNA 载体对 SHIV 复制抑制的评价

• 批准号: 7562014
• 负责人: Stephen E. Braun
• 金额: $ 5.2万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-05-01 至 2008-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7562014
• 关键词: Acquired Immunodeficiency Syndrome; Address; Clinical Trials; Collaborations; Computer Retrieval of Information on Scientific Projects Database; Evaluation; Experimental Models; Funding; Generations; Genes; Grant; HIV; HIV-1; Hematopoietic stem cells; Human; Immune; Institution; Life; Macaca; Modeling; Numbers; Polymerase; Reporting; Research; Research Personnel; Resources; Site; Small Interfering RNA; Source; Stem cells; Transfection; United States National Institutes of Health; Viral; Virus; gene therapy; in vivo; nonhuman primate; promoter; reconstitution; research study; simian human immunodeficiency virus; small hairpin RNA; vector

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Introduction of genes able to inhibit HIV replication into hematopoietic stem cells offers the potential for long-lived immune reconstitution. However, multiple ethical and practical considerations significantly constrain the ability to address basic questions regarding stem cell gene therapy for AIDS in human clinical trials. Experiments in nonhuman primates therefore offer the opportunity to rigorously address these issues in an in vivo experimental model. A number of recent reports have highlighted the potential utility of small interfering RNA (siRNA) molecules to inhibit viral replication. In collaboration with J. Rossi, we initiated inhibition studies of various SHIV strains with siRNA targeting Tat and Rev. We obtained two siRNA (one targeting both Tat and Rev and the other targeting only Rev) and a control sequence, each transcriptionally regulated by the polymerase III promoter U6. Using transient transfection both si(I) and si(II) strongly inhibited HIV-1 (as previously shown), SHIV 89.6p and SHIV Hxbc2 3.2P (viruses with homologous envelopes). Using a second generation construct expressing the site I and site II interfering RNAs as a hairpin sequence (shRNA) has demonstrated even more potent inhibition of SHIV replication by over 1000-fold. These encouraging results support further studies of the utility of siRNA to inhibit SHIV replication in vivo in the macaque model.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 引入能够抑制HIV复制到造血干细胞的基因提供了长期免疫重建的潜力。但是，多个道德和实际考虑因素显着限制了在人类临床试验中解决有关干细胞基因治疗的基本问题的能力。 因此，非人类灵长类动物的实验提供了在体内实验模型中严格解决这些问题的机会。 许多最近的报道突显了小型干扰RNA（siRNA）分子抑制病毒复制的潜在效用。 我们与J. Rossi合作，与靶向TAT和REV的siRNA启动了对各种SHIV菌株的抑制作用研究。我们获得了两个siRNA（一个针对TAT和REV，另一个针对Rev的siRNA）和一个控制序列，每个siRNA受聚合酶III启动子U6的转录调节。 使用瞬时转染Si（i）和Si（II）强烈抑制HIV-1（如前所述），SHIV 89.6P和SHIV HXBC2 3.2p（具有同源信封的病毒）。 使用表达位点I和站点II干扰RNA作为发夹序列（SHRNA）的第二代构造，已经显示出对SHIV复制的更有效抑制超过1000倍。 这些令人鼓舞的结果支持进一步研究siRNA在猕猴模型中抑制体内SHIV复制的实用性。