To employ gamma-peptide nucleic acid oligomers to interfere with viral replication

利用γ-肽核酸寡聚体干扰病毒复制

• 批准号: 10381536
• 负责人: Nara Lee
• 金额: $ 23.61万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-02 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10381536
• 关键词: Address; Affinity; Air; Animals; Antisense Oligonucleotide Therapy; Antisense Oligonucleotides; Antiviral resistance; Area; Binding; Binding Sites; Biochemical; Biological; Biological Assay; Biology; Biotin; Cell Culture Techniques; Cells; Cessation of life; Chemistry; Complementary RNA; Complex; Coupled; DNA; Databases; Development; Drug Kinetics; Epithelial Cells; Future; Generations; Genes; Genome; Genomic DNA; Genomics; Human; Hybrids; Infection; Influenza; Influenza A Virus, H1N1 Subtype; Influenza A Virus, H3N2 Subtype; Influenza A virus; Life; Life Cycle Stages; Liquid substance; Maps; Mediating; Microscope; Molecular Conformation; Monitor; Nucleoproteins; Nucleotides; Peptide Nucleic Acids; Peptides; Plague; Play; Polymerase; Population; Positioning Attribute; Production; Proteins; Quantitative Reverse Transcriptase PCR; RNA; RNA replication; Resolution; Ribonucleoproteins; Role; Site; Streptavidin; Temperature; Therapeutic; Time; United States; Universities; Vaccine Production; Vertebral column; Viral; Viral Genome; Virion; Virus; Virus Diseases; Virus Replication; Zoonoses; base; bronchial epithelium; chemical synthesis; crosslinking and immunoprecipitation sequencing; design; efficacy evaluation; egg; experimental study; fluorophore; in vivo; influenza infection; influenza virus vaccine; influenzavirus; melting; multidisciplinary; novel; nucleic acid analog; pandemic influenza; peptide analog; physiologic model; scaffold; seasonal influenza; uptake; vaccine efficacy; viral RNA; virology; water solubility

PROJECT SUMMARY Seasonal influenza A viruses (IAV) cause millions of infections and thousands of deaths each year in the US. Low vaccine efficacy and rapid antiviral resistance requires development of alternative therapeutic strategies to limit the burden of influenza on the population. The genome of IAV consists of eight negative- sense RNA segments that upon infection are first copied into positive-sense complementary RNA (cRNA), which in turn are replicated into genomic negative-sense viral RNA (vRNA) segments that will be packaged into progeny viruses. As the viral life cycle critically depends on template-mediated vRNA replication, an appealing approach to halt virus multiplication would be to place complementary antisense oligonucleotides (ASOs) as roadblocks on vRNA segments to stall viral polymerase progression. Our objective in this proposal is to examine the suitability of gamma-peptide nucleic acids (PNAs) as an antiviral against influenza virus replication. Gamma-PNAs are second-generation analogues of PNAs that show enhanced affinity, water solubility and biological activity when targeted to cellular RNA or genomic DNA. As this modified backbone enhances the hybridization affinity with target RNAs by significantly increasing the melting temperature of the resulting duplex, gamma-PNAs, unlike conventional DNA or RNA oligomers, could indeed function as stable roadblocks for viral polymerase. We have assembled an interdisciplinary team with expertise in the relevant areas, i.e. RNA biology, influenza virology, and PNA chemistry, to successfully complete the proposed experiments. Taken together, our study will lay the groundwork for future in vivo pharmacokinetic studies in animals.

项目总结 季节性甲型流感病毒(IAV)每年导致数百万人感染和数千人死亡 我们。疫苗的低效和快速的抗病毒耐药性要求开发替代疗法 限制流感对人口造成负担的战略。IAV的基因组由8个阴性- 当感染时首先复制到正义互补RNA(CRNA)中的正义RNA片段， 它们又被复制到基因组负义病毒RNA(VRNA)片段中，该片段将被包装 转化为子代病毒。由于病毒的生命周期严重依赖于模板介导的vRNA复制，因此 阻止病毒复制的有效方法是将互补的反义寡核苷酸 (ASOS)作为vRNA片段的障碍，以阻止病毒聚合酶的进展。 我们在这项建议中的目标是检查伽马肽核酸(PNA)作为一种 抗流感病毒复制的抗病毒药物。Gamma-PNA是PNA的第二代类似物， 当靶向细胞RNA或基因组DNA时，显示出更高的亲和力、水溶性和生物活性。 由于这种修饰的骨架通过显著增加 与传统的DNA或RNA低聚物不同，产生的双链，伽马-PNA的熔化温度可以 确实是病毒聚合酶的稳定障碍。我们已经组建了一个跨学科的团队， 在相关领域，即RNA生物学、流感病毒学和PNA化学方面的专业知识，以成功 完成建议的实验。综上所述，我们的研究将为未来在体内的研究奠定基础 动物体内的药代动力学研究。