title: Antisense Oligonucleotides Targeting Raf-1 Block Japanese Encephalitis Virus In Vitro and In Vivo.
abstract: Japanese encephalitis virus (JEV) infections represent a major health concern in Southeast Asia since no effective treatments are available. Recently, several reports have demonstrated that inhibition of certain host cell proteins prevents viral infection. Raf-1 kinase is a central component of many signaling pathways involved in normal cell growth and oncogenic transformation, and Ras/Raf/ERK signaling activation has been observed during viral infections (including JEV infection). In this study, Raf-1 was confirmed to be upregulated by JEV infection, which suggested that Raf-1 might be important for JEV infection and might be a target for novel anti-JEV drugs. To determine the role of Raf-1 during the JEV infection process, antisense oligonucleotides (ASODNs) were used to downregulate Raf-1 expression in JEV-infected baby hamster kidney (BHK-21) cells and African green monkey kidney (Vero) cells. From five ASODNs candidates tested, Raf-1-1 (Raf-1 antisense) significantly downregulated Raf-1 protein expression levels, significantly inhibited cytopathic effect (CPE) in cultured cells, and reduced JEV RNA levels in cell medium without affecting cell viability. Furthermore, it also demonstrated that ASODN Raf-1-1 possessed therapeutic effects by using a lethal JEV infection mouse model. In conclusion, data presented in this report demonstrated that ASODN Raf-1-1 could suppress Raf-1 protein and that Raf-1 inhibition suppressed JEV replication in vitro and in vivo. These data provided evidence for targeting Raf-1 in the development of novel anti-JEV therapies. In addition, Raf-1-1 represents potential drugs that can be adapted for treating JEV infections.
authors.affiliation: 1 Beijing Institute of Radiation Medicine , Beijing, People's Republic of China .|2 Tianjin Institute of Health and Environmental Medicine , Tianjin, People's Republic of China .
authors.forename: Li
authors.initials: L
authors.lastname: Zhang
authors.affiliation: 1 Beijing Institute of Radiation Medicine , Beijing, People's Republic of China .|3 Henan University of Traditional Chinese Medicine , Zhengzhou, People's Republic of China .
authors.forename: Qingjun
authors.initials: Q
authors.lastname: Li
authors.affiliation: 1 Beijing Institute of Radiation Medicine , Beijing, People's Republic of China .
authors.forename: Xiaoran
authors.initials: X
authors.lastname: Ding
authors.affiliation: 4 Department of Pharmacy, Peking Union Medical College Hospital , Chinese Academy of Medical Science and Peking Union Medical College, Beijing, People's Republic of China .
authors.forename: Bo
authors.initials: B
authors.lastname: Zhang
authors.affiliation: 1 Beijing Institute of Radiation Medicine , Beijing, People's Republic of China .
authors.forename: Qiling
authors.initials: Q
authors.lastname: Zhang
authors.affiliation: 1 Beijing Institute of Radiation Medicine , Beijing, People's Republic of China .|5 Shandong Analysis and Test Center , Shandong Academy of Science, Jinan, People's Republic of China .
authors.forename: Xinyan
authors.initials: X
authors.lastname: Qu
authors.affiliation: 1 Beijing Institute of Radiation Medicine , Beijing, People's Republic of China .
authors.forename: Yujia
authors.initials: Y
authors.lastname: Huo
authors.affiliation: 1 Beijing Institute of Radiation Medicine , Beijing, People's Republic of China .|3 Henan University of Traditional Chinese Medicine , Zhengzhou, People's Republic of China .
authors.forename: Jing
authors.initials: J
authors.lastname: Yang
authors.affiliation: 1 Beijing Institute of Radiation Medicine , Beijing, People's Republic of China .|3 Henan University of Traditional Chinese Medicine , Zhengzhou, People's Republic of China .
authors.forename: Shengqi
authors.initials: S
authors.lastname: Wang
mesh_terms: D000818:Animals; D002522:Chlorocebus aethiops; D003412:Cricetulus; D004195:Disease Models, Animal; D004664:Encephalitis Virus, Japanese / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D004672:Encephalitis, Japanese / Q000235:genetics / Q000401:mortality / Q000628:therapy* / Q000821:virology; D004847:Epithelial Cells / Q000473:pathology / Q000821:virology; D048049:Extracellular Signal-Regulated MAP Kinases / Q000235:genetics* / Q000378:metabolism; D005260:Female; D005786:Gene Expression Regulation; D054884:Host-Pathogen Interactions; D007274:Injections, Intraperitoneal; D051379:Mice; D008807:Mice, Inbred BALB C; D016376:Oligonucleotides, Antisense / Q000008:administration & dosage / Q000235:genetics* / Q000378:metabolism; D019908:Proto-Oncogene Proteins c-raf / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D015398:Signal Transduction; D016019:Survival Analysis; D014709:Vero Cells; D014779:Virus Replication; D018631:ras Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: D016376:Oligonucleotides, Antisense; D019908:Proto-Oncogene Proteins c-raf; D048049:Extracellular Signal-Regulated MAP Kinases; D018631:ras Proteins
keywords: JEV; Raf-1; antisense oligonucleotide