Pub Date : 2025-12-16DOI: 10.1016/j.antiviral.2025.106334
Dong Ji , Meng-Wen He , Wen-Chang Wang , Wei Han , Yan Chen , Yan Liu , Le Li , Xu-Yang Li , Yi-Fan Guo , Wu-Cai Yang , Zheng Dong , Chun-Yan Wang , Jing Xu , Lin Tan , George Lau , Yongping Yang
Low-level viremia (LLV) still occurs in some patients with chronic hepatitis B (CHB) after entecavir (ETV) treatment. We aimed to evaluate the effects of LLV on the adverse long-term outcomes. A secondary analysis of a multicenter prospective cohort study, consisting patients who underwent liver biopsy and received entecavir treatment, was conducted. LLV was defined as detectable HBV DNA levels (20–2000 IU/mL) at week 48 after the imitation of ETV treatment, maintained complete virological response (CVR) was defined as HBV DNA <20 IU/mL. The zero time was defined as the date of LLV diagnosis, the primary endpoint was the occurrence of HCC. The time-to-event analyses were performed using log-rank analysis, and multivariable COX regression. The inverse probability of treatment weighting and propensity score matching were used in the sensitivity analyses. In total, 766 patients were enrolled in the final analysis, and LLV was found in 182 patients (23.8 %). Patients with LLV had a significantly higher 7-year cumulative incidence of HCC (18.7 % vs. 8.1 %, p < 0.001) and fibrosis progression rate (17.5 % vs. 8.7 %, p = 0.017) than those with CVR. Multivariate analysis identified LLV as an independent risk factor associated with HCC (adjusted HR: 3.935; 95 % CI: 2.218–6.979, p < 0.001) and fibrosis progression (adjusted OR 5.342; 95 % CI: 1.630–17.480, p = 0.006). A nomogram incorporating LLV, age, PLT and liver cirrhosis was developed and validated for HCC risk prediction, demonstrating excellent performance with a C-index of 0.778. In conclusion, LLV significantly promoted HCC occurrence and fibrosis progression in CHB patients receiving anti-HBV treatment.
一些慢性乙型肝炎（CHB）患者在恩替卡韦（ETV）治疗后仍会发生低水平病毒血症（LLV）。我们的目的是评估LLV对不良长期预后的影响。对一项多中心前瞻性队列研究进行了二次分析，该研究包括接受肝活检并接受恩替卡韦治疗的患者。LLV定义为在模拟ETV治疗后第48周可检测到的HBV DNA水平（20-2000 IU/mL），维持完全病毒学应答（CVR）定义为HBV DNA
{"title":"Low-level viremia increases the risk of adverse long-term outcomes in entecavir-treated patients with chronic hepatitis B","authors":"Dong Ji , Meng-Wen He , Wen-Chang Wang , Wei Han , Yan Chen , Yan Liu , Le Li , Xu-Yang Li , Yi-Fan Guo , Wu-Cai Yang , Zheng Dong , Chun-Yan Wang , Jing Xu , Lin Tan , George Lau , Yongping Yang","doi":"10.1016/j.antiviral.2025.106334","DOIUrl":"10.1016/j.antiviral.2025.106334","url":null,"abstract":"<div><div>Low-level viremia (LLV) still occurs in some patients with chronic hepatitis B (CHB) after entecavir (ETV) treatment. We aimed to evaluate the effects of LLV on the adverse long-term outcomes. A secondary analysis of a multicenter prospective cohort study, consisting patients who underwent liver biopsy and received entecavir treatment, was conducted. LLV was defined as detectable HBV DNA levels (20–2000 IU/mL) at week 48 after the imitation of ETV treatment, maintained complete virological response (CVR) was defined as HBV DNA <20 IU/mL. The zero time was defined as the date of LLV diagnosis, the primary endpoint was the occurrence of HCC. The time-to-event analyses were performed using log-rank analysis, and multivariable COX regression. The inverse probability of treatment weighting and propensity score matching were used in the sensitivity analyses. In total, 766 patients were enrolled in the final analysis, and LLV was found in 182 patients (23.8 %). Patients with LLV had a significantly higher 7-year cumulative incidence of HCC (18.7 % vs. 8.1 %, <em>p</em> < 0.001) and fibrosis progression rate (17.5 % vs. 8.7 %, <em>p</em> = 0.017) than those with CVR. Multivariate analysis identified LLV as an independent risk factor associated with HCC (adjusted HR: 3.935; 95 % CI: 2.218–6.979, <em>p</em> < 0.001) and fibrosis progression (adjusted OR 5.342; 95 % CI: 1.630–17.480, <em>p</em> = 0.006). A nomogram incorporating LLV, age, PLT and liver cirrhosis was developed and validated for HCC risk prediction, demonstrating excellent performance with a C-index of 0.778. In conclusion, LLV significantly promoted HCC occurrence and fibrosis progression in CHB patients receiving anti-HBV treatment.</div></div>","PeriodicalId":8259,"journal":{"name":"Antiviral research","volume":"246 ","pages":"Article 106334"},"PeriodicalIF":4.0,"publicationDate":"2025-12-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145780009","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-10DOI: 10.1016/j.antiviral.2025.106325
Helong Feng , Junjie Yang , Liren Jiang , Xiaoyu Suo , Yuqiao Zhu , Yu Shang , Hongcai Wang , Zhe Zeng , Lun Yao , Qingping Luo , Guoyuan Wen
Newcastle disease (ND) is an important infectious disease in poultry, causing substantial economic losses in many countries. Vaccination with live vaccines is a common strategy for ND control, and farmers generally vaccinate multiple times each year because the current ND vaccines have a relatively short protection period. Here, we report that the NDV strain TS09-C, as an in ovo vaccine, provided long-lasting protection for at least 12 months against virulent NDV. TS09-C significantly reduced the tissue viral titer, alleviated histopathological lesions, and induced proliferation of CD8+ T cells after challenge. TS09-C in ovo group vs PBS group, 142 immune-related DEGs were screened based on RNA-Seq, including 18 up-regulated DEGs and 124 down-regulated DEGs. The innate immune pathways of these genes, as NOD-like, Toll-like, RIG-I-like, Cytokine-cytokine receptor interaction, and all genes (including pro-inflammatory cytokines IL-1B, IL-6, IL-18, etc) in these pathways were significantly downregulated. These results indicate that NDV strain TS09-C is a long-lasting protection vaccine candidate against ND, and the protection might be due to the reason that TS09-C induces the proliferation of CD8+ T cells and inhibits the inflammatory response.
{"title":"Newcastle disease virus TS09-C strain provides long-lasting protection against virulent strain after in-ovo immunization","authors":"Helong Feng , Junjie Yang , Liren Jiang , Xiaoyu Suo , Yuqiao Zhu , Yu Shang , Hongcai Wang , Zhe Zeng , Lun Yao , Qingping Luo , Guoyuan Wen","doi":"10.1016/j.antiviral.2025.106325","DOIUrl":"10.1016/j.antiviral.2025.106325","url":null,"abstract":"<div><div>Newcastle disease (ND) is an important infectious disease in poultry, causing substantial economic losses in many countries. Vaccination with live vaccines is a common strategy for ND control, and farmers generally vaccinate multiple times each year because the current ND vaccines have a relatively short protection period. Here, we report that the NDV strain TS09-C, as an <em>in ovo</em> vaccine, provided long-lasting protection for at least 12 months against virulent NDV. TS09-C significantly reduced the tissue viral titer, alleviated histopathological lesions, and induced proliferation of CD8<sup>+</sup> T cells after challenge. TS09-C <em>in ovo</em> group vs PBS group, 142 immune-related DEGs were screened based on RNA-Seq, including 18 up-regulated DEGs and 124 down-regulated DEGs. The innate immune pathways of these genes, as NOD-like, Toll-like, RIG-I-like, Cytokine-cytokine receptor interaction, and all genes (including pro-inflammatory cytokines IL-1B, IL-6, IL-18, etc) in these pathways were significantly downregulated. These results indicate that NDV strain TS09-C is a long-lasting protection vaccine candidate against ND, and the protection might be due to the reason that TS09-C induces the proliferation of CD8<sup>+</sup> T cells and inhibits the inflammatory response.</div></div>","PeriodicalId":8259,"journal":{"name":"Antiviral research","volume":"246 ","pages":"Article 106325"},"PeriodicalIF":4.0,"publicationDate":"2025-12-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145740493","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-09DOI: 10.1016/j.antiviral.2025.106326
Addison C. Hill , Madison B. Schank , Yi Zhang , Ning Sun , Ling Wang , Juan Zhao , Puja Banik , Jaeden S. Pyburn , Holly Orfield , Janet W. Lightner , Tabitha O. Leshaodo , Xiao Y. Wu , Shunbin Ning , Mohamed El Gazzar , Jonathan P. Moorman , Haitao Guo , Zhi Q. Yao
HBV infection is a global public health problem. The current treatment using nucleotide analogues (NA) can suppress viral replication but cannot eliminate HBV infection due to the persistence of covalently closed circular DNA (cccDNA), which sustains HBV replication and integration into the host cell genome and is refractory to NA treatment. CRISPR/Cas9 has been used to disrupt integrated HBV DNA and minichromosomal cccDNA for HBV suppression, but its expression and delivery require viral or non-viral vectors, which pose safety concerns for human application. We have previously reported the use of synthetic guide RNA (gRNA)/Cas9 ribonucleoprotein (RNP) as a non-viral formulation for HBV gene editing and viral suppression. To formulate highly effective CRISPR/Cas9 modalities for HBV gene therapy, here we designed additional gRNA/Cas9 RNPs and compared their antiviral efficacy in HBV-transfected as well as -infected cells. We found that two selected gRNA/Cas9 RNPs (gRNA5/Cas9, gRNA9/Cas9, and particularly their combinations) elicited the most potent antiviral efficacy, as evidenced by the significant inhibition of HBV DNA, RNA, and protein productions. DNA sequencing of the treated cells revealed moderate to high rates of insertion and deletion (indel) or knock-out (KO) mutations at the HBV target genes. Gene alignment analysis showed a high level of conservation for both gRNA5 and gRNA9 target sequences across major HBV genotypes, indicating that these CRISPR-based gene editing therapeutics have the potential to target different HBV strains worldwide. Thus, these synthetic gRNA/Cas9 RNPs represent promising novel therapeutics that can be developed and utilized for HBV gene disruption and viral eradication.
{"title":"Suppression of HBV replication and expression by CRISPR/Cas9 ribonucleoproteins","authors":"Addison C. Hill , Madison B. Schank , Yi Zhang , Ning Sun , Ling Wang , Juan Zhao , Puja Banik , Jaeden S. Pyburn , Holly Orfield , Janet W. Lightner , Tabitha O. Leshaodo , Xiao Y. Wu , Shunbin Ning , Mohamed El Gazzar , Jonathan P. Moorman , Haitao Guo , Zhi Q. Yao","doi":"10.1016/j.antiviral.2025.106326","DOIUrl":"10.1016/j.antiviral.2025.106326","url":null,"abstract":"<div><div>HBV infection is a global public health problem. The current treatment using nucleotide analogues (NA) can suppress viral replication but cannot eliminate HBV infection due to the persistence of covalently closed circular DNA (cccDNA), which sustains HBV replication and integration into the host cell genome and is refractory to NA treatment. CRISPR/Cas9 has been used to disrupt integrated HBV DNA and minichromosomal cccDNA for HBV suppression, but its expression and delivery require viral or non-viral vectors, which pose safety concerns for human application. We have previously reported the use of synthetic guide RNA (gRNA)/Cas9 ribonucleoprotein (RNP) as a non-viral formulation for HBV gene editing and viral suppression. To formulate highly effective CRISPR/Cas9 modalities for HBV gene therapy, here we designed additional gRNA/Cas9 RNPs and compared their antiviral efficacy in HBV-transfected as well as -infected cells. We found that two selected gRNA/Cas9 RNPs (gRNA5/Cas9, gRNA9/Cas9, and particularly their combinations) elicited the most potent antiviral efficacy, as evidenced by the significant inhibition of HBV DNA, RNA, and protein productions. DNA sequencing of the treated cells revealed moderate to high rates of insertion and deletion (indel) or knock-out (KO) mutations at the HBV target genes. Gene alignment analysis showed a high level of conservation for both gRNA5 and gRNA9 target sequences across major HBV genotypes, indicating that these CRISPR-based gene editing therapeutics have the potential to target different HBV strains worldwide. Thus, these synthetic gRNA/Cas9 RNPs represent promising novel therapeutics that can be developed and utilized for HBV gene disruption and viral eradication.</div></div>","PeriodicalId":8259,"journal":{"name":"Antiviral research","volume":"245 ","pages":"Article 106326"},"PeriodicalIF":4.0,"publicationDate":"2025-12-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145733830","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-05DOI: 10.1016/j.antiviral.2025.106324
Wenqiao Yang , Yuan Xu , Runmin Kang , Min Zhang , Yuting Zhang , Zeng Yang , Huiyuan Jing , Yilin Liu , Jiaxiang Li , Yongjie Ge , Haibo Feng , Long Zhou , Jiang Pi , Jie Liu
Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) remains a critical threat to the swine industry worldwide due to its rapid evolution and limited vaccine efficacy. In this study, we synthesized and characterized poly (allylamine hydrochloride)-selenium nanoparticles (PAH-Se NPs) and evaluated their antiviral activity against PRRSV in vitro and the underlying mechanism. PAH-Se NPs exhibited uniform spherical morphology with average hydrodynamic diameter of 118 nm and excellent colloidal stability with a zeta potential of +50.7 mV. Cytotoxicity assays demonstrated low toxicity of PAH-Se NPs in MARC-145 cells with a CC50 of 40.24 μg/mL, indicating favorable biocompatibility. Multiple complementary assays, including viral RNA quantification, cell viability restoration, plaque assays, N protein expression analyses, and indirect immunofluorescence assay, showed that PAH-Se NPs could significantly inhibit PRRSV replication in a dose-dependent manner, with an IC50 of 0.076 μg/mL. Moreover, this inhibition was equally effective against other sub-genotypes of PRRSV, including VR2332 and NADC30-like variants, as well as the DNA virus and bacteria, indicating the broad-spectrum anti-microbial properties of PAH-Se NPs. Mechanistic studies indicated that PAH-Se NPs target viral invasion and replication by down-regulating the host factor DDX5, which interacts with PRRSV Nsp9 to facilitate viral RNA synthesis, and suppressed PRRSV-induced reactive oxygen species (ROS) accumulation. This work emphasized PAH-Se NPs as a potent, low-toxicity antiviral candidate, offering promising potential for the development of alternative therapeutics against PRRSV and related viral infections.
{"title":"Poly (allylamine hydrochloride)-selenium nanoparticles inhibit porcine reproductive and respiratory syndrome virus by targeting DDX5 and reactive oxygen species","authors":"Wenqiao Yang , Yuan Xu , Runmin Kang , Min Zhang , Yuting Zhang , Zeng Yang , Huiyuan Jing , Yilin Liu , Jiaxiang Li , Yongjie Ge , Haibo Feng , Long Zhou , Jiang Pi , Jie Liu","doi":"10.1016/j.antiviral.2025.106324","DOIUrl":"10.1016/j.antiviral.2025.106324","url":null,"abstract":"<div><div>Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) remains a critical threat to the swine industry worldwide due to its rapid evolution and limited vaccine efficacy. In this study, we synthesized and characterized poly (allylamine hydrochloride)-selenium nanoparticles (PAH-Se NPs) and evaluated their antiviral activity against PRRSV in vitro and the underlying mechanism. PAH-Se NPs exhibited uniform spherical morphology with average hydrodynamic diameter of 118 nm and excellent colloidal stability with a zeta potential of +50.7 mV. Cytotoxicity assays demonstrated low toxicity of PAH-Se NPs in MARC-145 cells with a CC<sub>50</sub> of 40.24 μg/mL, indicating favorable biocompatibility. Multiple complementary assays, including viral RNA quantification, cell viability restoration, plaque assays, N protein expression analyses, and indirect immunofluorescence assay, showed that PAH-Se NPs could significantly inhibit PRRSV replication in a dose-dependent manner, with an IC<sub>50</sub> of 0.076 μg/mL. Moreover, this inhibition was equally effective against other sub-genotypes of PRRSV, including VR2332 and NADC30-like variants, as well as the DNA virus and bacteria, indicating the broad-spectrum anti-microbial properties of PAH-Se NPs. Mechanistic studies indicated that PAH-Se NPs target viral invasion and replication by down-regulating the host factor DDX5, which interacts with PRRSV Nsp9 to facilitate viral RNA synthesis, and suppressed PRRSV-induced reactive oxygen species (ROS) accumulation. This work emphasized PAH-Se NPs as a potent, low-toxicity antiviral candidate, offering promising potential for the development of alternative therapeutics against PRRSV and related viral infections.</div></div>","PeriodicalId":8259,"journal":{"name":"Antiviral research","volume":"245 ","pages":"Article 106324"},"PeriodicalIF":4.0,"publicationDate":"2025-12-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145699407","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Chronic hepatitis B virus (HBV) infection is a major health problem, affecting around 254 million people worldwide. Current treatments have side effects and can lead to resistance. Therefore, natural compounds derived from plants are being studied as potential antivirals. Isochlorogenic acid A (ICAA), which is derived from caffeoylquinic acid, exerts antiviral and hepatoprotective effects. The antioxidant effect of ICAA, triggered by upregulation of heme oxygenase 1 (HO-1), has been suggested as potential antiviral mechanism. However, the underlying mechanisms remain enigmatic. Our aim is to elucidate the mode of action of the antiviral effect of ICAA on HBV. Stable or transient transfected cells expressing HBV as well as HBV-infected cells were instrumental. (Sub)viral particles were characterized by biophysical and biochemical methods. Subcellular distribution of viral proteins was studied using confocal laser scanning microscopy. Viral genomes and transcripts were quantified by qPCR.
Treatment with ICAA decreased levels of HBV surface and e antigens (HBsAg and HBeAg), as well as viral transcripts, genomes and most important cccDNA. Furthermore, impaired virus assembly was evident from accumulation of naked capsids suggesting improper capsid formation and impaired envelopment. ICAA-dependent effects on HBV correlate with upregulation of HO-1 and modulation of intracellular ROS Our data indicate a possible link between changes in the intracellular ROS level and altered free -SH groups in viral structural proteins, possibly influencing proper disulphide bond formation and thereby assembly. In conclusion ICAA-dependent effects on HBV life cycle are based on several pillars as modulation of intracellular ROS and impaired morphogenesis and replication.
{"title":"Isochlorogenic acid A impairs hepatitis B virus replication by interference with various steps of hepatitis B virus life cycle involving HO-1-mediated ROS modulation","authors":"Giscard Wilfried Koyaweda , Mirco Glitscher , Anja Schollmeier , Daniela Bender , Eberhard Hildt","doi":"10.1016/j.antiviral.2025.106323","DOIUrl":"10.1016/j.antiviral.2025.106323","url":null,"abstract":"<div><div>Chronic hepatitis B virus (HBV) infection is a major health problem, affecting around 254 million people worldwide. Current treatments have side effects and can lead to resistance. Therefore, natural compounds derived from plants are being studied as potential antivirals. Isochlorogenic acid A (ICAA), which is derived from caffeoylquinic acid, exerts antiviral and hepatoprotective effects. The antioxidant effect of ICAA, triggered by upregulation of heme oxygenase 1 (HO-1), has been suggested as potential antiviral mechanism. However, the underlying mechanisms remain enigmatic. Our aim is to elucidate the mode of action of the antiviral effect of ICAA on HBV. Stable or transient transfected cells expressing HBV as well as HBV-infected cells were instrumental. (Sub)viral particles were characterized by biophysical and biochemical methods. Subcellular distribution of viral proteins was studied using confocal laser scanning microscopy. Viral genomes and transcripts were quantified by qPCR.</div><div>Treatment with ICAA decreased levels of HBV surface and e antigens (HBsAg and HBeAg), as well as viral transcripts, genomes and most important cccDNA. Furthermore, impaired virus assembly was evident from accumulation of naked capsids suggesting improper capsid formation and impaired envelopment. ICAA-dependent effects on HBV correlate with upregulation of HO-1 and modulation of intracellular ROS Our data indicate a possible link between changes in the intracellular ROS level and altered free -SH groups in viral structural proteins, possibly influencing proper disulphide bond formation and thereby assembly. In conclusion ICAA-dependent effects on HBV life cycle are based on several pillars as modulation of intracellular ROS and impaired morphogenesis and replication.</div></div>","PeriodicalId":8259,"journal":{"name":"Antiviral research","volume":"245 ","pages":"Article 106323"},"PeriodicalIF":4.0,"publicationDate":"2025-12-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145682331","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-01DOI: 10.1016/j.antiviral.2025.106322
John D. Sears , Konstantin I. Popov , Paul A. Sylvester , Rebekah Dickmander , Jennifer Diaz , Che-Kang Chang , Julia Huff , Wes Sanders , Nicholas A. Saba , Madeleine Sorensen , Adam M. Drobish , Nicholas A. May , Kevin Namitz , Julia Fecko , Neela H. Yennawar , Thomas E. Morrison , Alexander Tropsha , Mark T. Heise , Nathaniel J. Moorman
Emerging viruses pose an ongoing threat to human health. While certain viral families are common sources of outbreaks, predicting the specific virus within a family that will cause the next outbreak or pandemic is not possible, creating an urgent need for broad spectrum antiviral drugs that are effective against a wide array of related viral pathogens. However, broad spectrum drug development is hindered by the lack of detailed knowledge of compound binding sites that are structurally and functionally conserved between viral family members and are essential for virus replication. To overcome this limitation, we developed an in silico approach that combines AI-driven protein structure prediction, computational fragment soaking, multiple sequence alignment, and protein stability calculations to identify highly conserved target sites that are both solvent-accessible and conserved. We applied this approach to the Togaviridae family, which includes emerging pandemic disease threats such as chikungunya and Venezuelan equine encephalitis virus for which there are currently no approved antiviral therapies. Our analysis identified multiple solvent accessible and structurally conserved pockets in the alphavirus non-structural protein 2 (nsP2) protease domain, which is essential for processing the viral replicase proteins. Mutagenesis of key solvent accessible and conserved residues identified novel pockets that are essential for the replication of multiple alphaviruses, validating these pockets as potential antiviral target sites for nsP2 inhibitors. These findings highlight the potential of artificial intelligence-informed modeling for revealing functionally conserved, accessible pockets as a means of identifying potential target binding sites for broadly active direct acting antivirals.
{"title":"Novel computational pipeline to identify target sites for broad spectrum antiviral drugs","authors":"John D. Sears , Konstantin I. Popov , Paul A. Sylvester , Rebekah Dickmander , Jennifer Diaz , Che-Kang Chang , Julia Huff , Wes Sanders , Nicholas A. Saba , Madeleine Sorensen , Adam M. Drobish , Nicholas A. May , Kevin Namitz , Julia Fecko , Neela H. Yennawar , Thomas E. Morrison , Alexander Tropsha , Mark T. Heise , Nathaniel J. Moorman","doi":"10.1016/j.antiviral.2025.106322","DOIUrl":"10.1016/j.antiviral.2025.106322","url":null,"abstract":"<div><div>Emerging viruses pose an ongoing threat to human health. While certain viral families are common sources of outbreaks, predicting the specific virus within a family that will cause the next outbreak or pandemic is not possible, creating an urgent need for broad spectrum antiviral drugs that are effective against a wide array of related viral pathogens. However, broad spectrum drug development is hindered by the lack of detailed knowledge of compound binding sites that are structurally and functionally conserved between viral family members and are essential for virus replication. To overcome this limitation, we developed an in silico approach that combines AI-driven protein structure prediction, computational fragment soaking, multiple sequence alignment, and protein stability calculations to identify highly conserved target sites that are both solvent-accessible and conserved. We applied this approach to the <em>Togaviridae</em> family, which includes emerging pandemic disease threats such as chikungunya and Venezuelan equine encephalitis virus for which there are currently no approved antiviral therapies. Our analysis identified multiple solvent accessible and structurally conserved pockets in the alphavirus non-structural protein 2 (nsP2) protease domain, which is essential for processing the viral replicase proteins. Mutagenesis of key solvent accessible and conserved residues identified novel pockets that are essential for the replication of multiple alphaviruses, validating these pockets as potential antiviral target sites for nsP2 inhibitors. These findings highlight the potential of artificial intelligence-informed modeling for revealing functionally conserved, accessible pockets as a means of identifying potential target binding sites for broadly active direct acting antivirals.</div></div>","PeriodicalId":8259,"journal":{"name":"Antiviral research","volume":"245 ","pages":"Article 106322"},"PeriodicalIF":4.0,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145666763","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-11-29DOI: 10.1016/j.antiviral.2025.106321
Lung-Yi Mak , Anna SF. Lok
Functional cure has been proposed to be the treatment endpoint of cure therapies in chronic hepatitis B (CHB), yet it is rarely achieved with monotherapy of novel virus-targeting agents or immunomodulators. Although translation inhibitors – small interfering RNAs (siRNAs) and antisense oligonucleotides can produce marked decline in hepatitis B surface antigen (HBsAg) levels, the response is often not sustained and HBsAg seroclearance rarely occur after treatment cessation, suggesting that pharmacological reduction in HBsAg level may be insufficient in restoring HBV-specific immune response. Increasing number of studies have adopted the combination approach with virus-directing agent(s) plus immunomodulator(s). To date, the most effective regimen involves the concurrent or sequential use of siRNA with PEG-IFNα for 48 weeks, with resultant off-therapy HBsAg seroclearance rates approaching 30 %, and functional cure rates of up to 10 %. Other immunomodulators studied in combination with siRNA such as toll-like receptor agonists, therapeutic vaccines, monoclonal hepatitis B surface antibodies, and immune checkpoint inhibitors are less effective. Almost all studies included NUC and only a few evaluated protocolized NUC withdrawal; thus, few studies have truly evaluated functional cure. Low baseline HBsAg level is the most reliable predictor of HBsAg seroclearance, with many trials exclusively enrolling patients with HBsAg level <200 to <1000 IU/mL. While recent studies have shown promise, further research is needed to determine the optimal classes of drugs to combine, duration of use for each drug and whether they should be used concurrently or sequentially, to meet the desired goal of 30 % functional cure rate.
{"title":"Update on combination therapies against HBV in clinical investigations","authors":"Lung-Yi Mak , Anna SF. Lok","doi":"10.1016/j.antiviral.2025.106321","DOIUrl":"10.1016/j.antiviral.2025.106321","url":null,"abstract":"<div><div>Functional cure has been proposed to be the treatment endpoint of cure therapies in chronic hepatitis B (CHB), yet it is rarely achieved with monotherapy of novel virus-targeting agents or immunomodulators. Although translation inhibitors – small interfering RNAs (siRNAs) and antisense oligonucleotides can produce marked decline in hepatitis B surface antigen (HBsAg) levels, the response is often not sustained and HBsAg seroclearance rarely occur after treatment cessation, suggesting that pharmacological reduction in HBsAg level may be insufficient in restoring HBV-specific immune response. Increasing number of studies have adopted the combination approach with virus-directing agent(s) plus immunomodulator(s). To date, the most effective regimen involves the concurrent or sequential use of siRNA with PEG-IFNα for 48 weeks, with resultant off-therapy HBsAg seroclearance rates approaching 30 %, and functional cure rates of up to 10 %. Other immunomodulators studied in combination with siRNA such as toll-like receptor agonists, therapeutic vaccines, monoclonal hepatitis B surface antibodies, and immune checkpoint inhibitors are less effective. Almost all studies included NUC and only a few evaluated protocolized NUC withdrawal; thus, few studies have truly evaluated functional cure. Low baseline HBsAg level is the most reliable predictor of HBsAg seroclearance, with many trials exclusively enrolling patients with HBsAg level <200 to <1000 IU/mL. While recent studies have shown promise, further research is needed to determine the optimal classes of drugs to combine, duration of use for each drug and whether they should be used concurrently or sequentially, to meet the desired goal of 30 % functional cure rate.</div></div>","PeriodicalId":8259,"journal":{"name":"Antiviral research","volume":"245 ","pages":"Article 106321"},"PeriodicalIF":4.0,"publicationDate":"2025-11-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145647300","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
BK and JC Polyomavirus are closely related and establish persistence in infected subjects. Recent studies suggest that JC Polyomavirus replication prevents BK Polyomavirus-related pathologies in kidney transplant recipients. One potential mechanism of this competition could involve viral microRNAs cross-reacting, as they are highly homologous between species. In fact, bkv-miR-B1-3p is strictly identical to jcv-miR-J1-3p, whereas species-specific miRNAs bkv-miR-B1-5p and jcv-miR-J1-5p differ barely.
Early detection of jcv-miR-J1-5p in urine significantly reduces the risk of BK Polyomavirus DNAemia in kidney transplant recipients in a case-control study including 39 patients (odds ratio [95 % Confidence Interval] = 0.00 [0.00–0.65], p = 0.012). In vitro modeling revealed that prior infection with JC Polyomavirus reduces the ability of BK Polyomavirus to grow in immortalized human renal proximal tubular epithelial cells, without significant expression of JC Polyomavirus proteins. The JC Polyomavirus-specific miRNA jcv-miR-J1-5p was discovered to decrease BK Polyomavirus TAg mRNA expression, without affecting early genome replication, like the known regulatory effect of BK Polyomavirus-encoded bkv-miR-B1-3p and bkv-miR-B1-5p. An archetypal strain of JC Polyomavirus engineered to quench miRNA maturation did not inhibit BK Polyomavirus infection, unlike the wild-type strain, confirming that the inhibitory effect of JC Polyomavirus is due to miRNAs.
These results suggest that JC Polyomavirus-specific miRNA jcv-miR-J1-5p limits BK Polyomavirus infectivity and early TAg expression, with an intensity similar to BK Polyomavirus miRNAs. This mechanism might explain in vivo competition for viral replication between BK Polyomavirus and JC Polyomavirus infections.
BK和JC多瘤病毒密切相关，并在感染对象中建立持久性。最近的研究表明，JC多瘤病毒的复制可预防肾移植受者的BK多瘤病毒相关病理。这种竞争的一种潜在机制可能涉及病毒microrna的交叉反应，因为它们在物种之间高度同源。事实上，bkv-miR-B1-3p与jjv - mir - j1 -3p严格相同，而物种特异性mirna bkv-miR-B1-5p和jjv - mir - j1 -5p几乎没有区别。在一项包括39例患者的病例对照研究中，尿中早期检测jcv-miR-J1-5p可显著降低肾移植受者发生BK多瘤病毒dna血症的风险（优势比[95%置信区间]= 0.00 [0.00-0.65],p = 0.012）。体外模型显示，先前感染JC多瘤病毒可降低BK多瘤病毒在永生化人肾近端小管上皮细胞中的生长能力，但JC多瘤病毒蛋白的表达不显著。发现JC多瘤病毒特异性miRNA jcv-miR-J1-5p可降低BK多瘤病毒TAg mRNA表达，但不影响早期基因组复制，如已知的BK多瘤病毒编码的bkv-miR-B1-3p和bkv-miR-B1-5p的调节作用。与野生型菌株不同，JC多瘤病毒原型株抑制miRNA成熟并没有抑制BK多瘤病毒感染，这证实JC多瘤病毒的抑制作用是由miRNA引起的。这些结果表明，JC多瘤病毒特异性miRNA jcv-miR-J1-5p限制了BK多瘤病毒的感染性和早期TAg的表达，其强度与BK多瘤病毒miRNA相似。这一机制可能解释了BK多瘤病毒和JC多瘤病毒感染在体内的病毒复制竞争。
{"title":"JC polyomavirus-encoded miRNA jcv-miR-J1-5p downregulates BK polyomavirus infection","authors":"Baptiste Demey , Aurélien Aubry , Virginie Morel , Louison Collet , Catherine Francois , Sandrine Castelain , Francois Helle , Etienne Brochot","doi":"10.1016/j.antiviral.2025.106313","DOIUrl":"10.1016/j.antiviral.2025.106313","url":null,"abstract":"<div><div>BK and JC Polyomavirus are closely related and establish persistence in infected subjects. Recent studies suggest that JC Polyomavirus replication prevents BK Polyomavirus-related pathologies in kidney transplant recipients. One potential mechanism of this competition could involve viral microRNAs cross-reacting, as they are highly homologous between species. In fact, bkv-miR-B1-3p is strictly identical to jcv-miR-J1-3p, whereas species-specific miRNAs bkv-miR-B1-5p and jcv-miR-J1-5p differ barely.</div><div>Early detection of jcv-miR-J1-5p in urine significantly reduces the risk of BK Polyomavirus DNAemia in kidney transplant recipients in a case-control study including 39 patients (odds ratio [95 % Confidence Interval] = 0.00 [0.00–0.65], p = 0.012). <em>In vitro</em> modeling revealed that prior infection with JC Polyomavirus reduces the ability of BK Polyomavirus to grow in immortalized human renal proximal tubular epithelial cells, without significant expression of JC Polyomavirus proteins. The JC Polyomavirus-specific miRNA jcv-miR-J1-5p was discovered to decrease BK Polyomavirus TAg mRNA expression, without affecting early genome replication, like the known regulatory effect of BK Polyomavirus-encoded bkv-miR-B1-3p and bkv-miR-B1-5p. An archetypal strain of JC Polyomavirus engineered to quench miRNA maturation did not inhibit BK Polyomavirus infection, unlike the wild-type strain, confirming that the inhibitory effect of JC Polyomavirus is due to miRNAs.</div><div>These results suggest that JC Polyomavirus-specific miRNA jcv-miR-J1-5p limits BK Polyomavirus infectivity and early TAg expression, with an intensity similar to BK Polyomavirus miRNAs. This mechanism might explain <em>in vivo</em> competition for viral replication between BK Polyomavirus and JC Polyomavirus infections.</div></div>","PeriodicalId":8259,"journal":{"name":"Antiviral research","volume":"245 ","pages":"Article 106313"},"PeriodicalIF":4.0,"publicationDate":"2025-11-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145616823","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-11-20DOI: 10.1016/j.antiviral.2025.106312
Denis Pasqual , Ilaria Artusi , Michele Paccagnella , Giulia Sibille , Mattia Mirandola , Sofia Appelberg , Maria Francesca Priore , Maira Zorzan , Massimo E. Maffei , Paola De Benedictis , Claudia Del Vecchio , Alì Mirazimi , Giorgio Cozza , Giorgio Gribaudo , Cristiano Salata
The Ebola virus (EBOV) and the Rabies virus (RABV) are deadly infectious agents impacting human and animal health. Current prevention and control strategies mainly rely on vaccines and antibodies, highlighting the urgent need for effective, low-cost antivirals suitable for therapeutic options. Plant-derived bioactive compounds offer a promising natural source for such candidate antivirals. As a contribution to this antiviral approach, we have characterized the anti-EBOV and anti-RABV activity of a Cranberry extract (CE) endowed with a very high content of bioactive A2-type proanthocyanidin (PAC-A2). The CE inhibited the in vitro infection of both pseudoviruses expressing EBOV-GP or RABV-G glycoproteins and authentic EBOV and RABV. Attachment and entry assays revealed that the extract targets early phases of infection preventing attachment and entry. Noteworthy, synthetic PAC-A2 reproduced the antiviral activity observed with the whole CE. Mechanistic studies then revealed that the CE interacted directly with the ectodomain of EBOV-GP or the RABV-G, suggesting interference with their functions. In support to this hypothesis, fluorescence spectroscopy analysis showed a reduction in intrinsic fluorescence of both EBOV-GP and RABV-G after incubation with synthetic PAC-A2, thus confirming a direct interaction of the viral glycoproteins with PAC-A2. In silico docking simulations further sustained in vitro results by predicting the binding of PAC-A2 into the binding pocket of EBOV-GP and to the trimeric architecture of RABV-G.
Together, these results suggest this cranberry extract and bioactive PAC-A2 as potential candidates to be further develop as novel antiviral agents for the prevention of EBOV and RABV infections.
{"title":"The natural polyphenol proanthocyanidin A2 prevents the in vitro infection of Ebola virus and rabies virus by interfering with the early phases of the replication cycle","authors":"Denis Pasqual , Ilaria Artusi , Michele Paccagnella , Giulia Sibille , Mattia Mirandola , Sofia Appelberg , Maria Francesca Priore , Maira Zorzan , Massimo E. Maffei , Paola De Benedictis , Claudia Del Vecchio , Alì Mirazimi , Giorgio Cozza , Giorgio Gribaudo , Cristiano Salata","doi":"10.1016/j.antiviral.2025.106312","DOIUrl":"10.1016/j.antiviral.2025.106312","url":null,"abstract":"<div><div>The Ebola virus (EBOV) and the Rabies virus (RABV) are deadly infectious agents impacting human and animal health. Current prevention and control strategies mainly rely on vaccines and antibodies, highlighting the urgent need for effective, low-cost antivirals suitable for therapeutic options. Plant-derived bioactive compounds offer a promising natural source for such candidate antivirals. As a contribution to this antiviral approach, we have characterized the anti-EBOV and anti-RABV activity of a Cranberry extract (CE) endowed with a very high content of bioactive A2-type proanthocyanidin (PAC-A2). The CE inhibited the <em>in vitro</em> infection of both pseudoviruses expressing EBOV-GP or RABV-G glycoproteins and authentic EBOV and RABV. Attachment and entry assays revealed that the extract targets early phases of infection preventing attachment and entry. Noteworthy, synthetic PAC-A2 reproduced the antiviral activity observed with the whole CE. Mechanistic studies then revealed that the CE interacted directly with the ectodomain of EBOV-GP or the RABV-G, suggesting interference with their functions. In support to this hypothesis, fluorescence spectroscopy analysis showed a reduction in intrinsic fluorescence of both EBOV-GP and RABV-G after incubation with synthetic PAC-A2, thus confirming a direct interaction of the viral glycoproteins with PAC-A2. <em>In silico</em> docking simulations further sustained <em>in vitro</em> results by predicting the binding of PAC-A2 into the binding pocket of EBOV-GP and to the trimeric architecture of RABV-G.</div><div>Together, these results suggest this cranberry extract and bioactive PAC-A2 as potential candidates to be further develop as novel antiviral agents for the prevention of EBOV and RABV infections.</div></div>","PeriodicalId":8259,"journal":{"name":"Antiviral research","volume":"244 ","pages":"Article 106312"},"PeriodicalIF":4.0,"publicationDate":"2025-11-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145576257","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-11-20DOI: 10.1016/j.antiviral.2025.106310
Deyao Li , Yuxin Song , Danjuan Lu , Yanhua Zhang , Chenxiao Qu , Guiwen Guan , Yuexi Ma , Tianhao Mao , Liwei Zheng , Yi Wang , Wenhui Song , Ting Zhang , Fengmin Lu , Jing Ning , Xiangmei Chen
Elucidation of the regulation mechanism of hepatitis B virus (HBV) replication will provide potential targets for the development of novel anti-HBV therapeutics. It has been reported that the N6-methyladenosine (m6A) modification of HBV RNA plays a crucial role in the HBV life cycle. However, the mechanisms underlying the regulation of this modification remain incompletely understood. In this study, combining loss- and gain-of-function genetic analyses, we defined the role of IGF2BP1, an m6A reader, in facilitating HBV replication. Mechanistic studies revealed that IGF2BP1 stabilizes HBV RNAs primarily by binding to m6A-modified A1907 sites through its KH3-4 domain, thereby enhancing viral replication. Furthermore, targeted inhibition of IGF2BP1 by Cucurbitacin B, a small molecule inhibitor of IGF2BP1, was shown to inhibit HBV replication in vitro and in vivo. Taken together, these findings identify IGF2BP1 as a critical host regulator of HBV RNA stability through an m6A-dependent manner and targeted inhibition of IGF2BP1 effectively attenuates viral replication, providing a promising strategy for anti-HBV drug development.
{"title":"Targeted inhibition of IGF2BP1 effectively suppresses HBV replication via an m6A-dependent manner","authors":"Deyao Li , Yuxin Song , Danjuan Lu , Yanhua Zhang , Chenxiao Qu , Guiwen Guan , Yuexi Ma , Tianhao Mao , Liwei Zheng , Yi Wang , Wenhui Song , Ting Zhang , Fengmin Lu , Jing Ning , Xiangmei Chen","doi":"10.1016/j.antiviral.2025.106310","DOIUrl":"10.1016/j.antiviral.2025.106310","url":null,"abstract":"<div><div>Elucidation of the regulation mechanism of hepatitis B virus (HBV) replication will provide potential targets for the development of novel anti-HBV therapeutics. It has been reported that the N6-methyladenosine (m6A) modification of HBV RNA plays a crucial role in the HBV life cycle. However, the mechanisms underlying the regulation of this modification remain incompletely understood. In this study, combining loss- and gain-of-function genetic analyses, we defined the role of IGF2BP1, an m6A reader, in facilitating HBV replication. Mechanistic studies revealed that IGF2BP1 stabilizes HBV RNAs primarily by binding to m6A-modified A1907 sites through its KH3-4 domain, thereby enhancing viral replication. Furthermore, targeted inhibition of IGF2BP1 by Cucurbitacin B, a small molecule inhibitor of IGF2BP1, was shown to inhibit HBV replication <em>in vitro</em> and <em>in vivo</em>. Taken together, these findings identify IGF2BP1 as a critical host regulator of HBV RNA stability through an m6A-dependent manner and targeted inhibition of IGF2BP1 effectively attenuates viral replication, providing a promising strategy for anti-HBV drug development.</div></div>","PeriodicalId":8259,"journal":{"name":"Antiviral research","volume":"245 ","pages":"Article 106310"},"PeriodicalIF":4.0,"publicationDate":"2025-11-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145581632","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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