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

项目摘要 季节性甲型流感病毒（IAV）每年在美国造成数百万人感染和数千人死亡。 我们低疫苗效力和快速抗病毒耐药性需要开发替代治疗药物 限制流感对人口负担的战略。IAV的基因组由8个阴性- 感染后首先复制成正义互补RNA（cRNA）的正义RNA片段， 其又被复制成基因组负义病毒RNA（vRNA）片段， 转化为后代病毒。由于病毒的生命周期严重依赖于模板介导的vRNA复制， 阻止病毒增殖的吸引人的方法是将互补的反义寡核苷酸 （AS 0）作为vRNA区段上的路障以阻止病毒聚合酶进展。 我们在这项提案中的目标是检查γ-肽核酸（PNA）作为一种 抗流感病毒的复制。γ-PNA是PNA的第二代类似物， 当靶向细胞RNA或基因组DNA时显示增强的亲和力、水溶性和生物活性。 由于这种修饰的骨架通过显著增加与靶RNA的杂交而增强与靶RNA的杂交亲和力， 与传统的DNA或RNA寡聚物不同，所得双链体的解链温度，γ-PNA， 确实作为病毒聚合酶稳定路障。我们组建了一个跨学科的团队， 在相关领域的专业知识，即RNA生物学，流感病毒学和PNA化学，成功地 完成拟议的实验。综上所述，本研究为今后的体内实验奠定了基础 动物药代动力学研究。

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.