Bioactivity of Aptamers Targeted to HIV Reverse Transcriptase

针对 HIV 逆转录酶的适体的生物活性

• 批准号: 8074077
• 负责人: Margaret J Lange
• 金额: $ 5.13万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2013-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8074077
• 关键词: Animal Model; Anti-HIV Agents; Anti-HIV Therapy; Binding; Biological Assay; Cell Culture System; Cells; Clinical; Collaborations; Communities; Complex; DNA; Development; Drug resistance; Experimental Models; Future; Generations; Goals; HIV; HIV Infections; HIV drug resistance; Immune system; Laboratories; Methods; Missouri; Molecular Target; Nucleic Acids; Public Health; RNA; RNA-Directed DNA Polymerase; Research; Research Personnel; Research Proposals; Resistance; Resistance development; Role; Specialist; System; Technology; Therapeutic; Universities; Virus; Work; aptamer; combat; combinatorial; fitness; news; research study; resistance mutation; targeted delivery; weapons

DESCRIPTION (provided by applicant): HIV reverse transcriptase (RT) is required for HIV replication and remains a primary target for anti-HIV therapies. However, due to the rapid development of HIV resistance to anti-HIV drugs, it is necessary to develop new strategies to combat HIV infection. Nucleic acid aptamers are ssDNA or RNA molecules that bind selectively and tightly to specific molecular targets. Anti-RT aptamers have been demonstrated to significantly inhibit HIV RT in enzymatic assays. Some aptamers were shown to inhibit a diverse panel of HIV RTs (universalist) while others inhibited only specific RTs (specialists). Although a few aptamers have been demonstrated to inhibit HIV replication, the bioactivity of nucleic acid aptamers has not been thoroughly explored and the potential mechanisms for the generation of HIV resistance to aptamers have not been determined. The goals of this proposal are to define the bioactivity of DNA and RNA aptamers using both single-cycle HIV replication and replication-competent HIV assays. These experiments will compare universalists and specialists to define aptamer components promoting the most potent inhibition. In addition, the project will explore the potential of HIV resistance to DNA and RNA aptamers and determine the scope of resistance generated. In particular, the type of resistance generated from universalist versus specialist aptamers will be explored. Since universalist aptamers inhibit such a diverse number of RTs, it is hoped that it will be difficult for the virus to develop resistance to these aptamers and that resistance mutations will decrease the fitness of the virus. This proposal sets a firm background for future studies to explore aptamer delivery methods more applicable to a clinical setting and also to study aptamer effects on HIV replication in the context of more complex experimental models, such as a 3D cell culture system or an animal model to examine the role of the immune system in aptamer therapeutics. Relevance to Public Health: Approximately 10% of new HIV infections are drug resistant. DNA aptamers are a possible weapon against HIV and are highly stable and easily synthesized cheaply and efficiently using technology available worldwide, but delivery methods must be modified for clinical use. RNA aptamers are also attractive candidates for therapeutics and have several advantages in delivery over DNA aptamers but have not been studied sufficiently to identify the universal components needed for successful therapy. This project will investigate both aptamers in an effort to eventually couple the advantages into one therapeutic system. In addition to the potential for therapeutic application, the project will increase understanding of HIV resistance.

描述（由申请人提供）：HIV 逆转录酶 (RT) 是 HIV 复制所必需的，并且仍然是抗 HIV 治疗的主要靶点。然而，由于HIV对抗HIV药物的耐药性迅速发展，有必要制定新的策略来对抗HIV感染。核酸适体是选择性地紧密结合特定分子靶标的单链DNA或RNA分子。在酶测定中，抗 RT 适体已被证明可显着抑制 HIV RT。一些适体被证明可以抑制多种 HIV RT（通用型），而另一些适体仅抑制特定的 RT（专家型）。尽管一些适配体已被证明可以抑制HIV复制，但核酸适配体的生物活性尚未得到彻底探索，HIV对适配体产生耐药性的潜在机制尚未确定。该提案的目标是使用单周期 HIV 复制和复制能力 HIV 检测来定义 DNA 和 RNA 适体的生物活性。这些实验将比较通用主义者和专家，以确定促进最有效抑制的适体成分。此外，该项目还将探索HIV对DNA和RNA适体产生耐药性的潜力，并确定产生耐药性的范围。特别是，将探讨通用适体与专业适体产生的耐药类型。由于通用适体抑制如此多种RT，因此希望病毒难以对这些适体产生抗性，并且抗性突变将降低病毒的适应性。该提案为未来的研究奠定了坚实的背景，以探索更适用于临床环境的适体递送方法，并在更复杂的实验模型（例如3D细胞培养系统或动物模型）的背景下研究适体对HIV复制的影响，以检查免疫系统在适体治疗中的作用。与公共卫生的相关性：大约 10% 的新 HIV 感染具有耐药性。 DNA适体是一种可能的对抗HIV的武器，并且高度稳定，并且易于利用全球现有技术廉价且高效地合成，但必须修改递送方法以供临床使用。 RNA 适体也是有吸引力的治疗候选者，并且在递送方面比 DNA 适体具有多种优势，但尚未得到充分研究以鉴定成功治疗所需的通用成分。该项目将研究这两种适体，以期最终将其优点结合到一个治疗系统中。除了治疗应用的潜力外，该项目还将增进对艾滋病毒耐药性的了解。