Determinates of anti-HIV nucleic acid aptamer potency and resistance

抗HIV核酸适体效力和耐药性的测定

• 批准号: 7924282
• 负责人: Donald H Burke
• 金额: $ 6.08万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-22 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7924282
• 关键词: AIDS/HIV problem; Affinity; Amino Acid Sequence; Anti-HIV Agents; Antiviral Agents; Binding; Cells; Chemicals; Complex; Elements; Engineering; Evolution; Face; Future; Gene Therapy Agent; Genetic; Genetic Transcription; Goals; HIV; HIV-1; HIV-1 Reverse Transcriptase; In Vitro; Indium; Infection; Kinetics; Libraries; Measures; Modification; Molecular; Mutation; Nature; Nucleic Acids; RNA; RNA-Directed DNA Polymerase; Resistance; Structure; Toxic effect; Variant; Viral; Virus; Virus Diseases; aptamer; base; combat; design; drug resistant virus; gene therapy; high throughput screening; improved; in vitro activity; mutant; novel; novel therapeutics; pressure; public health relevance; response

DESCRIPTION (provided by applicant): New therapeutic strategies are needed to circumvent the rapid selection of drug resistant virus and the toxicity of current anti-HIV drugs. RNA aptamers targeted to the reverse transcriptase (RT) inhibit viral replication when expressed inside cells, and they offer great potential in future therapies against HIV/AIDS. There is little known about how the virus might evolve in the face of continual selection pressure, or about how the aptamers can be improved to circumvent potential resistance. The long-term goal of this project is to develop RNA aptamers as gene therapy agents that provide long-term antiviral protection against a rapidly-evolving virus. Our emphasis is on defining the influence of RT amino acid sequence variations on inhibition by aptamers, and on identifying new aptamer sequences and structures with improved inhibition, both in vitro and in cells. Aim 1 will determine the biophysical and structural basis for aptamer recognition and for differential inhibition by aptamers across phylogenetically diverse lentiviral RT's. Aim 2 will identify new aptamers with broad recognition and improved potency, and delineate the secondary structures associated with cross-clade enzymatic inhibition. Aim 3 examines inhibition of pseudotyped and replication-competent HIV-1 by intracellularly expressed aptamers, moving from optimization of the design elements that control expression, localization and inhibition to high throughput screens of aptamer and RT libraries. Aim 4 evaluates the de novo evolution of aptamer resistance with emphasis on establishing genetic threshold, resistance loci, and the molecular basis of resistance. PUBLIC HEALTH RELEVANCE This project will evaluate the potential for, and the nature of, HIV-1 resistance to nucleic acid aptamers-a promising class of molecules for eventual use in gene therapy strategies to combat HIV-1-and it will generate new aptamers that are less subject to escape mutations.

描述（由申请人提供）：需要新的治疗策略来规避耐药性病毒的快速选择和当前抗HIV药物的毒性。针对逆转录酶（RT）的RNA适体在细胞内表达时会抑制病毒复制，并且它们在针对HIV/AIDS的未来疗法中具有巨大的潜力。鲜为人知的病毒在面对持续选择压力的情况下如何发展，或者如何改善适体以规避潜在的抗性。该项目的长期目标是开发RNA适体作为基因治疗剂，可为快速发展的病毒提供长期抗病毒药物。我们的重点是定义RT氨基酸序列变化对适体抑制的影响，并在体外和细胞中鉴定具有改善抑制作用的新的适体序列和结构。 AIM 1将确定适体识别的生物物理和结构基础，以及适体在系统发育多样的慢病毒RT中的差异抑制。 AIM 2将确定具有广泛识别和提高效力的新适体，并描述与交叉酶抑制相关的二级结构。 AIM 3检查了通过细胞内表达的适体对假型和复制能力的HIV-1的抑制，从控制表达，定位和抑制的设计元素转变为适体和RT文库的高吞吐量筛选。 AIM 4评估了适体耐药性的从头演变，重点是建立遗传阈值，抗性基因座和抗性的分子基础。公共卫生相关性该项目将评估HIV-1对核酸适体的耐药性的潜力和性质 - 一种有希望的分子类，以最终用于基因治疗策略来对抗HIV-1不太容易逃脱突变。