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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要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。疫苗功效低下和抗病毒快速耐药需要开发替代疗法限制流感对人口负担的战略。 IAV 基因组由八个负有义RNA片段在感染后首先被复制成正义互补RNA（cRNA），进而被复制成基因组负义病毒 RNA (vRNA) 片段并进行包装转化为子代病毒。由于病毒生命周期关键取决于模板介导的 vRNA 复制，阻止病毒增殖的有吸引力的方法是放置互补的反义寡核苷酸（ASO）作为 vRNA 片段的障碍，阻止病毒聚合酶的进展。我们本提案的目标是检查 γ 肽核酸 (PNA) 作为抗流感病毒复制。 Gamma-PNA 是 PNA 的第二代类似物，当靶向细胞 RNA 或基因组 DNA 时，显示出增强的亲和力、水溶性和生物活性。由于这种修饰后的主链通过显着增加与目标 RNA 的杂交亲和力与传统的 DNA 或 RNA 寡聚物不同，所得双链体、γ-PNA 的解链温度可以确实起到了病毒聚合酶稳定路障的作用。我们组建了一支跨学科团队相关领域的专业知识，即 RNA 生物学、流感病毒学和 PNA 化学，以成功完成建议的实验。总而言之，我们的研究将为未来的体内研究奠定基础动物药代动力学研究。