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快速选择耐药病毒和当前抗艾滋病毒药物的毒性。针对逆转录酶(RT)的RNA适配子在细胞内表达时抑制病毒复制，它们在未来抗艾滋病毒/艾滋病的治疗中提供了巨大的潜力。对于病毒在持续的选择压力下可能会如何进化，或者如何改进适配子以规避潜在的耐药性，人们知之甚少。该项目的长期目标是开发RNA适配子作为基因治疗剂，为快速进化的病毒提供长期的抗病毒保护。我们的重点是确定RT氨基酸序列变异对适配子抑制的影响，并在体外和细胞内鉴定具有改进抑制作用的新的适配子序列和结构。目的1将确定适配子识别的生物物理和结构基础，以及在系统发育不同的慢病毒RT中识别适配子和差异抑制的生物物理和结构基础。目的2将寻找具有广泛识别和改进效力的新的适配子，并描绘与交叉型酶抑制相关的二级结构。目的3研究细胞内表达的适体对假型和复制能力HIV-1的抑制作用，从优化控制表达、定位和抑制的设计元件转移到高通量筛选适体和RT文库。目的4评价适配子抗性的从头进化，重点是建立遗传阈值、抗性位点和抗性的分子基础。公共卫生意义该项目将评估HIV-1对核酸适配子的抗药性的潜力和性质--核酸适配子是一类很有希望最终用于抗击HIV-1的基因治疗策略的分子--它将产生不太容易发生逃逸突变的新适配子。