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: 8266475
• 负责人: Margaret J Lange
• 金额: $ 5.39万
• 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/8266475
• 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复制需要HIV逆转录酶（RT），并且仍然是抗HIV疗法的主要目标。但是，由于对抗HIV药物的抗HIV耐药性的快速发展，有必要制定新的策略来打击HIV感染。核酸适体是ssDNA或RNA分子，它们与特定分子靶标有选择性结合。已经证明抗RT适体在酶试验中显着抑制HIV RT。一些适体被证明可以抑制各种艾滋病毒RT（普遍主义者），而另一些适体仅抑制特定的RT（专家）。尽管已经证明了一些适体可以抑制HIV复制，但尚未彻底探索核酸适体的生物活性，并且尚未确定对适体产生HIV耐药性的潜在机制。该提案的目标是使用单周期HIV复制和竞争能力的HIV分析来定义DNA和RNA适体的生物活性。这些实验将比较普遍主义者和专家，以定义促进最有效抑制的适体组件。此外，该项目将探索HIV对DNA和RNA适体的耐药性，并确定产生的抗性范围。特别是，将探索由普遍主义者与专家适体产生的阻力类型。由于普遍主义的适体抑制了这种不同数量的RT，因此希望病毒难以发展对这些适体的抗性，并且耐药突变会降低病毒的适应性。该提案为将来的研究设定了一个牢固的背景，以探索更适用于临床环境的适体输送方法，并在更复杂的实验模型的背景下研究适合对HIV复制的影响，例如3D细胞培养系统或动物模型，以检查适当疗法中免疫系统的作用。与公共卫生有关：大约10％的新艾滋病毒感染具有抗药性。 DNA Aptamer是一种可能针对艾滋病毒的武器，使用全球可用的技术易于廉价且易于合成，但必须修改交付方法以供临床使用。 RNA Aptamers也是治疗疗法的有吸引力的候选者，并且在交付中比DNA适体具有多个优势，但尚未进行足够的研究以识别成功治疗所需的通用组件。该项目将调查两个适合学家，以最终将优势融入一个治疗系统中。除了具有治疗应用的潜力外，该项目还将增加对艾滋病毒抗性的理解。