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逆转录酶（RT）是HIV复制所必需的，仍然是抗HIV治疗的主要靶点。然而，由于艾滋病毒对抗艾滋病毒药物的耐药性迅速发展，有必要制定新的战略来对抗艾滋病毒感染。核酸适体是选择性地且紧密地结合至特定分子靶标的ssDNA或RNA分子。已证明抗RT适体在酶测定中显著抑制HIV RT。一些适体被证明可以抑制多种HIV RT（普遍主义者），而另一些适体仅抑制特定的RT（专家）。虽然一些适体已被证明可以抑制HIV复制，但核酸适体的生物活性尚未被彻底探索，并且尚未确定HIV对适体产生抗性的潜在机制。该提案的目标是使用单循环HIV复制和复制能力HIV检测来确定DNA和RNA适体的生物活性。这些实验将比较普适论者和专家，以确定促进最有效抑制的适体组分。此外，该项目还将探索艾滋病毒对DNA和RNA适体的耐药性潜力，并确定产生耐药性的范围。特别是，将探讨普适适体与专门适体产生的耐药性类型。由于普适适体抑制如此多样数量的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。这一提议为未来的研究奠定了坚实的基础，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（如3D细胞培养系统或动物模型）中研究适体对HIV复制的影响，以研究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%的新艾滋病毒感染者具有耐药性。DNA适体是一种可能的抗艾滋病毒武器，并且高度稳定，易于使用世界范围内可用的技术廉价有效地合成，但必须修改输送方法以供临床使用。RNA适体也是治疗剂的有吸引力的候选物，并且在递送方面具有优于DNA适体的几个优点，但尚未充分研究以鉴定成功治疗所需的通用组分。该项目将研究两种适体，以最终将其优势结合到一个治疗系统中。除了潜在的治疗应用外，该项目还将增加对艾滋病毒耐药性的了解。