Background: Japanese encephalitis virus (JEV), a mosquito-borne flavivirus, remains a leading cause of viral encephalitis. Current management is largely supportive, with no specific antivirals. This study evaluated the antiviral efficacy and mechanism of action of CC-90009 against JEV in vitro and in vivo. Methods: Five targeted protein degraders (TPDs) were screened for anti-JEV activity in the human neuroblastoma cell line SH-SY5Y. Time-of-addition, binding, and endocytosis assays were used to delineate the phase of action of CC-90009, a cereblon (CRBN) E3 ligase modulator (CELMoD) and molecular glue degrader. Small interfering RNA knockdown and co-immunoprecipitation (Co-IP) confirmed targets essential for its antiviral effects. The broad-spectrum activity of CC-90009 against other mosquito-borne viruses was also evaluated. In vivo efficacy was tested in a murine JEV model. Results: Of the five TPDs tested, only CC-90009 significantly inhibited JEV infection in SH-SY5Y cells, acting during both viral entry and post-entry phases without reducing adsorbed or internalised virions. CC-90009 reduced JEV RNA and non-structural protein accumulation. Knockdown of G1-to-S phase transition 1 (GSPT1), a key target of CC-90009, suppressed JEV infection and translation; Co-IP confirmed GSPT1 interaction with JEV non-structural protein 5 (NS5). CC-90009 disrupted JEV translation and replication by inducing proteasomal degradation of the GSPT1/NS5 complex, further demonstrating its broad-spectrum antiviral activity through the effective inhibition of West Nile virus and chikungunya virus. In vivo, it protected mice from JEV-induced mortality, reducing viral load, antigen levels, and brain pathology. Conclusions: CC-90009 exerts potent anti-JEV activity both in vitro and in vivo by inducing proteasomal degradation of the GSPT1/NS5 complex, thereby disrupting viral translation and replication. This targeted protein degradation strategy represents a novel host-directed antiviral approach with promising therapeutic potential against mosquito-borne viral encephalitis.

Chikungunya fever (CHIKF), which is caused by the Chikungunya virus (CHIKV), has rapidly spread across the globe in recent years, leading to its listing as a public health concern by the World Health Organization. In 2019, the first local outbreak of Asian-type CHIKF was reported in Xishuangbanna, Yunnan Province, China, with 88 CHIKV nucleic acid-positive cases detected from clinical specimens. To further investigate the biological characteristics of the virus strain responsible for this outbreak, we reconstructed the 625D6h strain using reverse genetics and tested its growth kinetics in different cell lines. The results showed a strong replication capacity in the Aedes albopictus C6/36 insect cell line but a weaker one in mammalian cell lines. The virus’s high replication capacity in mosquito cells is an interesting phenotype that warrants further study to determine if it influences vector adaptation and transmission dynamics in endemic settings. The study also found that two DHODH inhibitors, ML390 and vidofludimus, could effectively inhibit CHIKV replication in vitro. The infectious clone created in this study provides a useful tool for studying the recently prevalent Asian-type strain in China, supporting the subsequent development of prevention and treatment methods.

Chikungunya fever (CHIKF) is caused by the Chikungunya virus (CHIKV). In recent years, it has rapidly spread globally and has been listed as a public health concern by the World Health Organization. In 2019, the first local outbreak of Asian-type CHIKF was reported in Xishuangbanna, Yunnan Province, China, with 88 CHIKV nucleic acid-positive cases detected from clinical specimens. To further investigate the biological characteristics of the virus strain responsible for this outbreak, we rescued the 625D6h strain using reverse genetics and tested its growth kinetics in different cell lines. The results showed that the 625D6h strain had strong replication capacity in the Aedes albopictus C6/36 insect cell line but weaker replication capacity in mammalian cell lines. Its high replication capacity under high mosquito vector adaptability may have contributed to the outbreak in 2019. The study also found that two DHODH inhibitors, ML390 and vidofludimus, could effectively inhibit CHIKV replication. The infectious clone created in this study provides a useful tool for studying the recently prevalent Asian-type strain in China and supports the subsequent development of prevention and treatment methods.

Background: Dengue fever is one of the most widely distributed vector-borne infectious diseases globally, prevalent in tropical and subtropical regions. An estimated 3.9 billion people in 128 countries are at risk of infection globally, and the 2024 outbreak was the most severe. As a tropical region, Hainan Province in China serves as a significant endemic area for dengue fever, where epidemic prevention and control remain critical. Objective: To analyze the transmission dynamics and pathogen variation characteristics of the 2024 dengue fever outbreak in Hainan Province. Methods: Acute-phase serum samples from 18 confirmed dengue cases during the 2024 outbreak in Hainan Province were collected and serotyped using TaqMan real-time PCR. The viral envelope (E) gene and representative whole genomes were sequenced and compared with domestic reference strains to assess epidemic trends. Phylogenetic and molecular clock analyses were performed based on E gene and full-genome sequences to infer spatiotemporal transmission dynamics. Sequence alignment and homology modeling were used to identify E gene mutations and evaluate their potential effects on E protein functional domains and viral pathogenicity. Results: Among the 18 samples, 17 were dengue virus (DENV)-1, and 1 was DENV-3. Phylogenetic analysis revealed that the primary strains in this outbreak were highly homologous to those circulating in Guangdong Province; however, two strains exhibited significant genetic differences, suggesting possible independent introduction. Additionally, two unique DENV-1 E gene mutations (D147N and S338L) were identified, which we hypothesize may be associated with viral characteristics, subject to further functional verification. Conclusion: This study systematically reveals the epidemiological characteristics of the 2024 dengue fever outbreak and provides a scientific basis for formulating local control strategies and a research foundation for further exploring the molecular-level pathogenicity and transmissibility of DENV.