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: 7802678
• 负责人: Margaret J Lange
• 金额: $ 4.76万
• 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/7802678
• 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(申请人提供)：艾滋病毒逆转录酶(RT)是艾滋病毒复制所必需的，并且仍然是抗艾滋病毒治疗的主要靶点。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。核酸适配子是选择性地与特定分子靶标紧密结合的单链DNA或RNA分子。在酶分析中，反RT适配子已被证明能显著抑制HIV RT。一些适配子被证明能抑制一组不同的HIV RT(普适主义者)，而其他适体只抑制特定的RT(专家)。尽管少数适配子已被证明能抑制HIV复制，但核酸适配子的生物活性还没有得到彻底的研究，导致HIV对适配子产生耐药性的潜在机制还没有确定。这项建议的目标是使用单周期艾滋病毒复制和复制能力艾滋病毒检测来确定DNA和RNA适配子的生物活性。这些实验将对通用主义者和专家进行比较，以确定促进最有效抑制的适体成分。此外，该项目将探索艾滋病毒对DNA和RNA适配子的耐药性潜力，并确定产生耐药性的范围。特别是，将探索由普遍性适配子与专业适配子产生的抗性类型。由于普遍性适配子抑制了如此多的RT，因此希望病毒对这些适配子很难产生耐药性，并且耐药性突变将降低病毒的适合度。这一建议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输送方法，并在更复杂的实验模型的背景下研究适体对艾滋病毒复制的影响，例如3D细胞培养系统或动物模型，以检查免疫系统在适体疗法中的作用。与公共卫生的相关性：大约10%的新艾滋病毒感染具有抗药性。DNA适配子是对抗艾滋病毒的一种可能的武器，它高度稳定，利用世界各地现有的技术廉价而高效地容易合成，但必须改进给药方法以用于临床。RNA适配子也是有吸引力的治疗候选药物，在递送方面比DNA适配子有几个优势，但还没有得到充分的研究，无法确定成功治疗所需的通用成分。该项目将研究这两种适体，以努力最终将这两种优势结合到一个治疗系统中。除了治疗应用的潜力外，该项目还将增加对艾滋病毒耐药性的了解。