ANTIVIRAL EFFICACY OF OLIGONUCLEOTIDES TO HIV-1 POLYPURINE-RICH SEQUENCES

寡核苷酸对富含 HIV-1 多嘌呤序列的抗病毒功效

• 批准号: 5200817
• 负责人: K L POFFENBERGER
• 金额: --
• 依托单位: BUREAU OF HEALTH PLANNING AND RESOURCES DEVELOPMENT
• 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/5200817
• 关键词: HIV infections; antiAIDS agent; antiviral agents; genome; human immunodeficiency virus 1; human tissue; nucleic acid inhibitor; nucleic acid structure; oligonucleotides; pyrimidine nucleotides; tissue /cell culture; triple helix; virus DNA; virus RNA; virus antigen; virus genetics; virus replication

Synthetic oligonucleotides designed to affect HIV pathogenesis by sequence-specific interactions with viral nucleic acid have shown promise as candidates for antiviral therapy. We have been exploring the comparative efficacy of oligonucleotide treatment (aimed at different stages in the viral replicative cycle) of HIV-1 infected cells in vitro. The family of oligos we have used target the same polypurine-rich sequences, but carry different modifications such that they may function as triple helix (TH), antisense (AS), or nonspecific inhibitors. Treatment of infected cells with ONs designed to form a Triple Helix (TH) with the target HIV sequence significantly decreased p24 antigen levels as well as decreasing the amounts of viral DNA and RNA relative to untreated cells. Treatment of infected cells with AS ONs also inhibited HIV expression, but in a more limited and less sequence specific manner. The oligos were rapidly taken up by the cells, entering the nuclei within 10 minutes and maintaining detectable, non-degraded levels for up to 48 hours. These oligos showed some interaction with their target sequences in in vitro assays, but the association was not stable. We have continued these studies with TH oligos carrying different modifications designed to 1) increase the stability of the TH structure, 2) provide more resistance to cellular nucleases, and 3) enhance the antiviral activity of the ON - target interaction. We have also begun studies to investigate the primary mode of action in ON inhibition of viral replication. These studies have been conducted in tissue culture cell lines (H9 cells) and with primary normal human peripheral blood lymphocytes (PBLs), using laboratory and primary isolates.

设计通过以下方式影响HIV发病机制的合成寡核苷酸 与病毒核酸的序列特异性相互作用已经显示出希望 作为抗病毒治疗的候选者。 我们一直在探索 寡核苷酸治疗的比较功效（针对不同的 病毒复制周期中的阶段）。 我们使用的寡核苷酸家族靶向相同的富含多嘌呤的 序列，但携带不同的修饰，使得它们可以发挥功能， 作为三螺旋（TH）、反义（AS）或非特异性抑制剂。 用设计成形成三重阻碍（TH）的ON处理感染的细胞 HIV靶序列显著降低了p24抗原水平 以及减少病毒DNA和RNA的量， 未经处理的细胞 用AS ONs处理感染的细胞也抑制了 HIV表达，但以更有限和更少序列特异性的方式。 寡核苷酸迅速被细胞吸收，进入细胞核内 10分钟并保持可检测的、非降解水平长达48 小时 这些寡核苷酸与它们的靶序列表现出一定的相互作用 在体外试验中，但协会是不稳定的。我们有 用携带不同修饰的TH寡核苷酸继续这些研究 设计成1）增加TH结构的稳定性，2）提供 对细胞核酸酶的抗性更强，3）增强抗病毒性 靶向相互作用的活性。 我们还开始研究， 研究ON抑制病毒的主要作用模式 复制的 这些研究已经在组织培养细胞中进行， 细胞系（H9细胞）和原代正常人外周血 淋巴细胞（PBL），使用实验室和初级分离株。