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Human norovirus is the leading cause of acute gastroenteritis worldwide, affecting every year 685 million people. Norovirus outbreaks are associated with very significant economic losses, with an estimated societal cost of 60 billion USD per year. Despite this, no therapeutic options or vaccines are currently available to treat or prevent this infection. An antiviral therapy that can be used as treatment and as a prophylactic measure in the case of outbreaks is urgently needed. We previously described the computer-aided design and synthesis of novel small-molecule agents able to inhibit the replication of human norovirus in cell-based systems. These compounds are non-nucleoside inhibitors of the viral polymerase and are characterized by a terminal para-substituted phenyl group connected to a central phenyl ring by an amide-thioamide linker, and a terminal thiophene ring. Here we describe new modifications of these scaffolds focused on exploring the role of the substituent at the para position of the terminal phenyl ring and on removing the thioamide portion of the amide-thioamide linker, to further explore structure-activity relationships (SARs) and improve antiviral properties. According to three to four-step synthetic routes, we prepared thirty novel compounds, which were then evaluated against the replication of both murine (MNV) and human (HuNoV) norovirus in cells. Derivatives in which the terminal phenyl group has been replaced by an unsubstituted benzoxazole or indole, and the thioamide component of the amide-thioamide linker has been removed, showed promising results in inhibiting HuNoV replication at low micromolar concentrations. Particularly, compound 28 was found to have an EC50 against HuNoV of 0.9 µM. Although the most active novel derivatives were also associated with an increased cytotoxicity in the human cell line, these compounds represent a very promising starting point for the development of new analogues with reduced cytotoxicity and improved selectivity indexes. In addition, the experimental biological data have been used to create an initial 3D quantitative structure-activity relationship model, which could be used to guide the future design of novel potential anti-norovirus agents.
Salvatore Ferla; Carmine Varricchio; William Knight; Pui Kei Ho; Fabiana Saporito; Beatrice Tropea; Giulio Fagan; Ben Matthew Flude; Federica Bevilacqua; Nanci Santos-Ferreira; Jana Van Dycke; Johan Neyts; Andrea Brancale; Joana Rocha-Pereira; Marcella Bassetto. Structure–Activity Relationship Studies on Novel Antiviral Agents for Norovirus Infections. Microorganisms 2021, 9, 1795 .
AMA StyleSalvatore Ferla, Carmine Varricchio, William Knight, Pui Kei Ho, Fabiana Saporito, Beatrice Tropea, Giulio Fagan, Ben Matthew Flude, Federica Bevilacqua, Nanci Santos-Ferreira, Jana Van Dycke, Johan Neyts, Andrea Brancale, Joana Rocha-Pereira, Marcella Bassetto. Structure–Activity Relationship Studies on Novel Antiviral Agents for Norovirus Infections. Microorganisms. 2021; 9 (9):1795.
Chicago/Turabian StyleSalvatore Ferla; Carmine Varricchio; William Knight; Pui Kei Ho; Fabiana Saporito; Beatrice Tropea; Giulio Fagan; Ben Matthew Flude; Federica Bevilacqua; Nanci Santos-Ferreira; Jana Van Dycke; Johan Neyts; Andrea Brancale; Joana Rocha-Pereira; Marcella Bassetto. 2021. "Structure–Activity Relationship Studies on Novel Antiviral Agents for Norovirus Infections." Microorganisms 9, no. 9: 1795.
Many species of the order Bunyavirales contain potentially fatal viruses that lack effective medical countermeasures and are therefore collectively a major public health threat. Here, we describe a cell-based assay using Bunyamwera virus (BUNV)-mCherry to identify and characterize new antiviral molecules against bunyaviruses. BUNV is the type species for the genus Orthobunyavirus and has been reported to cause mild symptoms in humans, such as fever, joint pain, and rash. One major benefit of using our fluorescence-based assay over classical CPE-based assays is the fact that the antiviral effect of the tested compounds and their effect on the cell viability can be determined within the same assay well. For that reason, this type of assay could significantly advance our preclinical efforts towards finding new antiviral molecules against bunyaviruses.
Sebastiaan ter Horst; Winston Chiu; Johan Neyts; Joana Rocha-Pereira. Screening and in vitro antiviral assessment of small molecules against fluorescent protein-expressing Bunyamwera virus in a cell-based assay using high-content imaging. 2021, 29, 1 .
AMA StyleSebastiaan ter Horst, Winston Chiu, Johan Neyts, Joana Rocha-Pereira. Screening and in vitro antiviral assessment of small molecules against fluorescent protein-expressing Bunyamwera virus in a cell-based assay using high-content imaging. . 2021; 29 ():1.
Chicago/Turabian StyleSebastiaan ter Horst; Winston Chiu; Johan Neyts; Joana Rocha-Pereira. 2021. "Screening and in vitro antiviral assessment of small molecules against fluorescent protein-expressing Bunyamwera virus in a cell-based assay using high-content imaging." 29, no. : 1.
Acute gastroenteritis caused by virus has a major impact on public health worldwide in terms of morbidity, mortality, and economic burden. The main culprits are rotaviruses, noroviruses, sapoviruses, astroviruses, and enteric adenoviruses. Currently, there are no antiviral drugs available for the prevention or treatment of viral gastroenteritis. Here, we describe the antivirals that were identified as having in vitro and/or in vivo activity against these viruses, originating from in silico design or library screening, natural sources or being repurposed drugs. We also highlight recent advances in model systems available for this (hard to cultivate) group of viruses, such as organoid technologies, and that will facilitate antiviral studies as well as fill some of current knowledge gaps that hamper the development of highly efficient therapies against gastroenteric viruses.
Nanci Santos-Ferreira; Jana Van Dycke; Johan Neyts; Joana Rocha-Pereira. Current and Future Antiviral Strategies to Tackle Gastrointestinal Viral Infections. Microorganisms 2021, 9, 1599 .
AMA StyleNanci Santos-Ferreira, Jana Van Dycke, Johan Neyts, Joana Rocha-Pereira. Current and Future Antiviral Strategies to Tackle Gastrointestinal Viral Infections. Microorganisms. 2021; 9 (8):1599.
Chicago/Turabian StyleNanci Santos-Ferreira; Jana Van Dycke; Johan Neyts; Joana Rocha-Pereira. 2021. "Current and Future Antiviral Strategies to Tackle Gastrointestinal Viral Infections." Microorganisms 9, no. 8: 1599.
We have recently established that human norovirus (HuNoV) replicates efficiently in zebrafish larvae after inoculation of a clinical sample into the yolk, providing a simple and robust in vivo system in which to study HuNoV. In this Protocol Extension, we present a detailed description of virus inoculation by microinjection, subsequent daily monitoring and harvesting of larvae, followed by viral RNA quantification. This protocol can be used to study viral replication of genogroup (G)I and GII HuNoVs in vivo within 3–4 d. Additionally, we describe how to evaluate the in vivo antiviral effect and toxicity of small molecules using HuNoV-infected zebrafish larvae, in multi-well plates and without the need for specific formulations. This constitutes a great advantage for drug discovery efforts, as no specific antivirals or vaccines currently exist to treat or prevent norovirus gastroenteritis. This Protocol Extension details the use of zebrafish larvae as a simple and robust in vivo system for studying human norovirus infection, enabling evaluation of the antiviral effects and toxicities of small molecules.
Jana Van Dycke; Arno Cuvry; Jan Knickmann; Annelii Ny; Sebastian Rakers; Stefan Taube; Peter de Witte; Johan Neyts; Joana Rocha-Pereira. Infection of zebrafish larvae with human norovirus and evaluation of the in vivo efficacy of small-molecule inhibitors. Nature Protocols 2021, 16, 1830 -1849.
AMA StyleJana Van Dycke, Arno Cuvry, Jan Knickmann, Annelii Ny, Sebastian Rakers, Stefan Taube, Peter de Witte, Johan Neyts, Joana Rocha-Pereira. Infection of zebrafish larvae with human norovirus and evaluation of the in vivo efficacy of small-molecule inhibitors. Nature Protocols. 2021; 16 (4):1830-1849.
Chicago/Turabian StyleJana Van Dycke; Arno Cuvry; Jan Knickmann; Annelii Ny; Sebastian Rakers; Stefan Taube; Peter de Witte; Johan Neyts; Joana Rocha-Pereira. 2021. "Infection of zebrafish larvae with human norovirus and evaluation of the in vivo efficacy of small-molecule inhibitors." Nature Protocols 16, no. 4: 1830-1849.
Human norovirus is the main cause of viral gastroenteritis, resulting annually in ∼ 700 million infections and 200,000 deaths, of whom most are children <5 years. Mouse norovirus-infected macrophages are the most widely used in vitro system to screen and characterize the antiviral effect of norovirus-targeting small molecules. We have previously established antiviral assays using this system, identified novel inhibitors and performed additional studies in order to have a first insight into their mechanism of action. As potent and safe anti-norovirus small molecules are urgently needed, we here describe the detailed protocol for a set of assays that will allow the identification of novel norovirus inhibitors.
Jana Van Dycke; Jasper Rymenants; Johan Neyts; Joana Rocha-Pereira. Assessment of the anti-norovirus activity in cell culture using the mouse norovirus: Identification of active compounds. Antiviral Chemistry and Chemotherapy 2021, 29, 1 .
AMA StyleJana Van Dycke, Jasper Rymenants, Johan Neyts, Joana Rocha-Pereira. Assessment of the anti-norovirus activity in cell culture using the mouse norovirus: Identification of active compounds. Antiviral Chemistry and Chemotherapy. 2021; 29 ():1.
Chicago/Turabian StyleJana Van Dycke; Jasper Rymenants; Johan Neyts; Joana Rocha-Pereira. 2021. "Assessment of the anti-norovirus activity in cell culture using the mouse norovirus: Identification of active compounds." Antiviral Chemistry and Chemotherapy 29, no. : 1.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) rapidly spread around the globe after its emergence in Wuhan in December 2019. With no specific therapeutic and prophylactic options available, the virus has infected millions of people of which more than half a million succumbed to the viral disease, COVID-19. The urgent need for an effective treatment together with a lack of small animal infection models has led to clinical trials using repurposed drugs without preclinical evidence of their in vivo efficacy. We established an infection model in Syrian hamsters to evaluate the efficacy of small molecules on both infection and transmission. Treatment of SARS-CoV-2−infected hamsters with a low dose of favipiravir or hydroxychloroquine with(out) azithromycin resulted in, respectively, a mild or no reduction in virus levels. However, high doses of favipiravir significantly reduced infectious virus titers in the lungs and markedly improved lung histopathology. Moreover, a high dose of favipiravir decreased virus transmission by direct contact, whereas hydroxychloroquine failed as prophylaxis. Pharmacokinetic modeling of hydroxychloroquine suggested that the total lung exposure to the drug did not cause the failure. Our data on hydroxychloroquine (together with previous reports in macaques and ferrets) thus provide no scientific basis for the use of this drug in COVID-19 patients. In contrast, the results with favipiravir demonstrate that an antiviral drug at nontoxic doses exhibits a marked protective effect against SARS-CoV-2 in a small animal model. Clinical studies are required to assess whether a similar antiviral effect is achievable in humans without toxic effects.
Suzanne J. F. Kaptein; Sofie Jacobs; Lana Langendries; Laura Seldeslachts; Sebastiaan ter Horst; Laurens Liesenborghs; Bart Hens; Valentijn Vergote; Elisabeth Heylen; Karine Barthelemy; Elke Maas; Carolien De Keyzer; Lindsey Bervoets; Jasper Rymenants; Tina Van Buyten; Xin Zhang; Rana Abdelnabi; Juanita Pang; Rachel Williams; Hendrik Jan Thibaut; Kai Dallmeier; Robbert Boudewijns; Jens Wouters; Patrick Augustijns; Nick Verougstraete; Christopher Cawthorne; Judith Breuer; Caroline Solas; Birgit Weynand; Pieter Annaert; Isabel Spriet; Greetje Vande Velde; Johan Neyts; Joana Rocha-Pereira; Leen Delang. Favipiravir at high doses has potent antiviral activity in SARS-CoV-2−infected hamsters, whereas hydroxychloroquine lacks activity. Proceedings of the National Academy of Sciences 2020, 117, 26955 -26965.
AMA StyleSuzanne J. F. Kaptein, Sofie Jacobs, Lana Langendries, Laura Seldeslachts, Sebastiaan ter Horst, Laurens Liesenborghs, Bart Hens, Valentijn Vergote, Elisabeth Heylen, Karine Barthelemy, Elke Maas, Carolien De Keyzer, Lindsey Bervoets, Jasper Rymenants, Tina Van Buyten, Xin Zhang, Rana Abdelnabi, Juanita Pang, Rachel Williams, Hendrik Jan Thibaut, Kai Dallmeier, Robbert Boudewijns, Jens Wouters, Patrick Augustijns, Nick Verougstraete, Christopher Cawthorne, Judith Breuer, Caroline Solas, Birgit Weynand, Pieter Annaert, Isabel Spriet, Greetje Vande Velde, Johan Neyts, Joana Rocha-Pereira, Leen Delang. Favipiravir at high doses has potent antiviral activity in SARS-CoV-2−infected hamsters, whereas hydroxychloroquine lacks activity. Proceedings of the National Academy of Sciences. 2020; 117 (43):26955-26965.
Chicago/Turabian StyleSuzanne J. F. Kaptein; Sofie Jacobs; Lana Langendries; Laura Seldeslachts; Sebastiaan ter Horst; Laurens Liesenborghs; Bart Hens; Valentijn Vergote; Elisabeth Heylen; Karine Barthelemy; Elke Maas; Carolien De Keyzer; Lindsey Bervoets; Jasper Rymenants; Tina Van Buyten; Xin Zhang; Rana Abdelnabi; Juanita Pang; Rachel Williams; Hendrik Jan Thibaut; Kai Dallmeier; Robbert Boudewijns; Jens Wouters; Patrick Augustijns; Nick Verougstraete; Christopher Cawthorne; Judith Breuer; Caroline Solas; Birgit Weynand; Pieter Annaert; Isabel Spriet; Greetje Vande Velde; Johan Neyts; Joana Rocha-Pereira; Leen Delang. 2020. "Favipiravir at high doses has potent antiviral activity in SARS-CoV-2−infected hamsters, whereas hydroxychloroquine lacks activity." Proceedings of the National Academy of Sciences 117, no. 43: 26955-26965.
Several fatal bunyavirus infections lack specific treatment. Here, we show that diketo acids engage a panel of bunyavirus cap-snatching endonucleases, inhibit their catalytic activity and reduce viral replication of a taxonomic representative in vitro. Specifically, the non-salt form of L-742,001 and its derivatives exhibited EC50 values between 5.6 to 6.9 μM against a recombinant BUNV-mCherry virus. Structural analysis and molecular docking simulations identified traits of both the class of chemical entities and the viral target that could help the design of novel, more potent molecules for the development of pan-bunyavirus antivirals.
Yaiza Fernández-García; Sebastiaan ter Horst; Marcella Bassetto; Andrea Brancale; Johan Neyts; Dominga Rogolino; Mario Sechi; Mauro Carcelli; Stephan Günther; Joana Rocha-Pereira. Diketo acids inhibit the cap-snatching endonuclease of several Bunyavirales. Antiviral Research 2020, 183, 104947 .
AMA StyleYaiza Fernández-García, Sebastiaan ter Horst, Marcella Bassetto, Andrea Brancale, Johan Neyts, Dominga Rogolino, Mario Sechi, Mauro Carcelli, Stephan Günther, Joana Rocha-Pereira. Diketo acids inhibit the cap-snatching endonuclease of several Bunyavirales. Antiviral Research. 2020; 183 ():104947.
Chicago/Turabian StyleYaiza Fernández-García; Sebastiaan ter Horst; Marcella Bassetto; Andrea Brancale; Johan Neyts; Dominga Rogolino; Mario Sechi; Mauro Carcelli; Stephan Günther; Joana Rocha-Pereira. 2020. "Diketo acids inhibit the cap-snatching endonuclease of several Bunyavirales." Antiviral Research 183, no. : 104947.
SARS-CoV-2 rapidly spread around the globe after its emergence in Wuhan in December 2019. With no specific therapeutic and prophylactic options available, the virus was able to infect millions of people. To date, close to half a million patients succumbed to the viral disease, COVID-19. The high need for treatment options, together with the lack of small animal models of infection has led to clinical trials with repurposed drugs before any preclinicalin vivoevidence attesting their efficacy was available. We used Syrian hamsters to establish a model to evaluate antiviral activity of small molecules in both an infection and a transmission setting. Upon intranasal infection, the animals developed high titers of SARS-CoV-2 in the lungs and pathology similar to that observed in mild COVID-19 patients. Treatment of SARS-CoV-2-infected hamsters with favipiravir or hydroxychloroquine (with and without azithromycin) resulted in respectively a mild or no reduction in viral RNA and infectious virus. Micro-CT scan analysis of the lungs showed no improvement compared to non-treated animals, which was confirmed by histopathology. In addition, both compounds did not prevent virus transmission through direct contact and thus failed as prophylactic treatments. By modelling the PK profile of hydroxychloroquine based on the trough plasma concentrations, we show that the total lung exposure to the drug was not the limiting factor. In conclusion, we here characterized a hamster infection and transmission model to be a robust model for studyingin vivoefficacy of antiviral compounds. The information acquired using hydroxychloroquine and favipiravir in this model is of critical value to those designing (current and) future clinical trials. At this point, the data here presented on hydroxychloroquine either alone or combined with azithromycin (together with previously reportedin vivodata in macaques and ferrets) provide no scientific basis for further use of the drug in humans.
Suzanne Jf Kaptein; Sofie Jacobs; Lana Langendries; Laura Seldeslachts; Sebastiaan Ter Horst; Laurens Liesenborghs; Bart Hens; Valentijn Vergote; Elisabeth Heylen; Elke Maas; Carolien De Keyzer; Lindsey Bervoets; Jasper Rymenants; Tina Van Buyten; Hendrik Jan Thibaut; Kai Dallmeier; Robbert Boudewijns; Jens Wouters; Patrick Augustijns; Nick Verougstraete; Christopher Cawthorne; Birgit Weynand; Pieter Annaert; Isabel Spriet; Greetje Vande Velde; Johan Neyts; Joana Rocha-Pereira; Leen Delang. Antiviral treatment of SARS-CoV-2-infected hamsters reveals a weak effect of favipiravir and a complete lack of effect for hydroxychloroquine. 2020, 1 .
AMA StyleSuzanne Jf Kaptein, Sofie Jacobs, Lana Langendries, Laura Seldeslachts, Sebastiaan Ter Horst, Laurens Liesenborghs, Bart Hens, Valentijn Vergote, Elisabeth Heylen, Elke Maas, Carolien De Keyzer, Lindsey Bervoets, Jasper Rymenants, Tina Van Buyten, Hendrik Jan Thibaut, Kai Dallmeier, Robbert Boudewijns, Jens Wouters, Patrick Augustijns, Nick Verougstraete, Christopher Cawthorne, Birgit Weynand, Pieter Annaert, Isabel Spriet, Greetje Vande Velde, Johan Neyts, Joana Rocha-Pereira, Leen Delang. Antiviral treatment of SARS-CoV-2-infected hamsters reveals a weak effect of favipiravir and a complete lack of effect for hydroxychloroquine. . 2020; ():1.
Chicago/Turabian StyleSuzanne Jf Kaptein; Sofie Jacobs; Lana Langendries; Laura Seldeslachts; Sebastiaan Ter Horst; Laurens Liesenborghs; Bart Hens; Valentijn Vergote; Elisabeth Heylen; Elke Maas; Carolien De Keyzer; Lindsey Bervoets; Jasper Rymenants; Tina Van Buyten; Hendrik Jan Thibaut; Kai Dallmeier; Robbert Boudewijns; Jens Wouters; Patrick Augustijns; Nick Verougstraete; Christopher Cawthorne; Birgit Weynand; Pieter Annaert; Isabel Spriet; Greetje Vande Velde; Johan Neyts; Joana Rocha-Pereira; Leen Delang. 2020. "Antiviral treatment of SARS-CoV-2-infected hamsters reveals a weak effect of favipiravir and a complete lack of effect for hydroxychloroquine." , no. : 1.
Introductory paragraphSince the emergence of SARS-CoV-2 causing COVID-19, the world is being shaken to its core with numerous hospitalizations and hundreds of thousands of deaths. In search for key targets of effective therapeutics, robust animal models mimicking COVID-19 in humans are urgently needed. Here, we show that productive SARS-CoV-2 infection in the lungs of mice is limited and restricted by early type I interferon responses. In contrast, we show that Syrian hamsters are highly permissive to SARS- CoV-2 and develop bronchopneumonia and a strong inflammatory response in the lungs with neutrophil infiltration and edema. Moreover, we identify an exuberant innate immune response as a key player in pathogenesis, in which STAT2 signaling plays a dual role, driving severe lung injury on the one hand, yet restricting systemic virus dissemination on the other. Finally, we assess SARS-CoV- 2-induced lung pathology in hamsters by micro-CT alike used in clinical practice. Our results reveal the importance of STAT2-dependent interferon responses in the pathogenesis and virus control during SARS-CoV-2 infection and may help rationalizing new strategies for the treatment of COVID-19 patients.
Robbert Boudewijns; Hendrik Jan Thibaut; Suzanne J. F. Kaptein; Rong Li; Valentijn Vergote; Laura Seldeslachts; Carolien De Keyzer; Lindsey Bervoets; Sapna Sharma; Johan Van Weyenbergh; Laurens Liesenborghs; Ji Ma; Sander Jansen; Dominique Van Looveren; Thomas Vercruysse; Dirk Jochmans; Xinyu Wang; Erik Martens; Kenny Roose; Dorien De Vlieger; Bert Schepens; Tina Van Buyten; Sofie Jacobs; Yanan Liu; Joan Martí-Carreras; Bert Vanmechelen; Tony Wawina-Bokalanga; Leen Delang; Joana Rocha-Pereira; Lotte Coelmont; Winston Chiu; Pieter Leyssen; Elisabeth Heylen; Dominique Schols; Lanjiao Wang; Lila Close; Jelle Matthijnssens; Marc Van Ranst; Veerle Compernolle; Georg Schramm; Koen Van Laere; Xavier Saelens; Nico Callewaert; Ghislain Opdenakker; Piet Maes; Birgit Weynand; Christopher Cawthorne; Greetje Vande Velde; Zhongde Wang; Johan Neyts; Kai Dallmeier. STAT2 signaling as double-edged sword restricting viral dissemination but driving severe pneumonia in SARS-CoV-2 infected hamsters. 2020, 1 .
AMA StyleRobbert Boudewijns, Hendrik Jan Thibaut, Suzanne J. F. Kaptein, Rong Li, Valentijn Vergote, Laura Seldeslachts, Carolien De Keyzer, Lindsey Bervoets, Sapna Sharma, Johan Van Weyenbergh, Laurens Liesenborghs, Ji Ma, Sander Jansen, Dominique Van Looveren, Thomas Vercruysse, Dirk Jochmans, Xinyu Wang, Erik Martens, Kenny Roose, Dorien De Vlieger, Bert Schepens, Tina Van Buyten, Sofie Jacobs, Yanan Liu, Joan Martí-Carreras, Bert Vanmechelen, Tony Wawina-Bokalanga, Leen Delang, Joana Rocha-Pereira, Lotte Coelmont, Winston Chiu, Pieter Leyssen, Elisabeth Heylen, Dominique Schols, Lanjiao Wang, Lila Close, Jelle Matthijnssens, Marc Van Ranst, Veerle Compernolle, Georg Schramm, Koen Van Laere, Xavier Saelens, Nico Callewaert, Ghislain Opdenakker, Piet Maes, Birgit Weynand, Christopher Cawthorne, Greetje Vande Velde, Zhongde Wang, Johan Neyts, Kai Dallmeier. STAT2 signaling as double-edged sword restricting viral dissemination but driving severe pneumonia in SARS-CoV-2 infected hamsters. . 2020; ():1.
Chicago/Turabian StyleRobbert Boudewijns; Hendrik Jan Thibaut; Suzanne J. F. Kaptein; Rong Li; Valentijn Vergote; Laura Seldeslachts; Carolien De Keyzer; Lindsey Bervoets; Sapna Sharma; Johan Van Weyenbergh; Laurens Liesenborghs; Ji Ma; Sander Jansen; Dominique Van Looveren; Thomas Vercruysse; Dirk Jochmans; Xinyu Wang; Erik Martens; Kenny Roose; Dorien De Vlieger; Bert Schepens; Tina Van Buyten; Sofie Jacobs; Yanan Liu; Joan Martí-Carreras; Bert Vanmechelen; Tony Wawina-Bokalanga; Leen Delang; Joana Rocha-Pereira; Lotte Coelmont; Winston Chiu; Pieter Leyssen; Elisabeth Heylen; Dominique Schols; Lanjiao Wang; Lila Close; Jelle Matthijnssens; Marc Van Ranst; Veerle Compernolle; Georg Schramm; Koen Van Laere; Xavier Saelens; Nico Callewaert; Ghislain Opdenakker; Piet Maes; Birgit Weynand; Christopher Cawthorne; Greetje Vande Velde; Zhongde Wang; Johan Neyts; Kai Dallmeier. 2020. "STAT2 signaling as double-edged sword restricting viral dissemination but driving severe pneumonia in SARS-CoV-2 infected hamsters." , no. : 1.
Recent evidence indicates that viral components of the microbiota can contribute to intestinal homeostasis and protection from local inflammatory or infectious insults. However, host-derived mechanisms that maintain tolerance to the virome remain largely unknown. Here, we use colonization with the model commensal murine norovirus (MNV CR6) to interrogate host-directed mechanisms of viral tolerance, and show that STAT1 is a central coordinator of tolerance following CR6 colonization. STAT1 restricts CR6 replication to the intestinal tract, prevents systemic viral-induced tissue damage and disease, and regulates antiviral CD4+ and CD8+ T cell responses. In contrast to systemic viral pathogens that drive T cell mediated immunopathology in STAT1-deficient mice, our data indicates that loss of CD4+ or CD8+ T cells and their associated effector functions has no effect on CR6-induced disease. However, therapeutic administration of an antiviral compound to limit viral replication prevented viral-induced tissue damage and death despite ongoing dysregulated antiviral T cell responses. Collectively, our data uncouple the requirement for STAT1-mediated regulation of antiviral T cell responses from innate immune-mediated restriction of viral replication that is necessary for intestinal viral tolerance.
Heather A Filyk; Andrew J Sharon; Nicolette M Fonseca; Rachel L Simister; Wallace Yuen; Blair K Hardman; Hannah G Robinson; Jung Hee Seo; Joana Rocha-Pereira; Ian Welch; Johan Neyts; Sean A Crowe; Lisa C Osborne. STAT1-dependent tolerance of intestinal viral infection. 2020, 1 .
AMA StyleHeather A Filyk, Andrew J Sharon, Nicolette M Fonseca, Rachel L Simister, Wallace Yuen, Blair K Hardman, Hannah G Robinson, Jung Hee Seo, Joana Rocha-Pereira, Ian Welch, Johan Neyts, Sean A Crowe, Lisa C Osborne. STAT1-dependent tolerance of intestinal viral infection. . 2020; ():1.
Chicago/Turabian StyleHeather A Filyk; Andrew J Sharon; Nicolette M Fonseca; Rachel L Simister; Wallace Yuen; Blair K Hardman; Hannah G Robinson; Jung Hee Seo; Joana Rocha-Pereira; Ian Welch; Johan Neyts; Sean A Crowe; Lisa C Osborne. 2020. "STAT1-dependent tolerance of intestinal viral infection." , no. : 1.
Human noroviruses (HuNoVs) are the most common cause of foodborne illness, with a societal cost of $60 billion and 219,000 deaths/year. The lack of robust small animal models has significantly hindered the understanding of norovirus biology and the development of effective therapeutics. Here we report that HuNoV GI and GII replicate to high titers in zebrafish (Danio rerio) larvae; replication peaks at day 2 post infection and is detectable for at least 6 days. The virus (HuNoV GII.4) could be passaged from larva to larva two consecutive times. HuNoV is detected in cells of the hematopoietic lineage and the intestine, supporting the notion of a dual tropism. Antiviral treatment reduces HuNoV replication by >2 log10, showing that this model is suited for antiviral studies. Zebrafish larvae constitute a simple and robust replication model that will largely facilitate studies of HuNoV biology and the development of antiviral strategies. Human norovirus (HuNoV) is the number one agent of viral gastroenteritis worldwide. It can infect people of all age groups, resulting in 700 million infections and 219,000 deaths each year. Outbreaks of acute HuNoV gastroenteritis occur often, but chronic infections also happen in people with immune deficiencies. Despite its clinical relevance, studying the virus has been a decades-long challenge because no simple and efficient cultivation system existed. This has recently started to change; we here contribute with an important step forward by establishing an in vivo model system to study HuNoV replication using zebrafish larvae. We inject a HuNoV in the yolk (food reserve) of 3-day-old zebrafish larvae, which results in high virus titers up to 6 days after inoculation. We could detect the virus in the tissues of the infected larvae by a variety of techniques including histology, which showed us in which organs the virus is present. Importantly, by adding an antiviral to the water in which the larvae swim, we could significantly reduce the virus levels in the larvae. This means that testing small molecules and developing the first antiviral therapy for HuNoV will be much easier from hereon.
Jana Van Dycke; Annelii Ny; Nádia Conceição-Neto; Jan Maes; Myra Hosmillo; Arno Cuvry; Ian Goodfellow; Tatiane C. Nogueira; Erik Verbeken; Jelle Matthijnssens; Peter de Witte; Johan Neyts; Joana Rocha-Pereira. A robust human norovirus replication model in zebrafish larvae. PLOS Pathogens 2019, 15, e1008009 .
AMA StyleJana Van Dycke, Annelii Ny, Nádia Conceição-Neto, Jan Maes, Myra Hosmillo, Arno Cuvry, Ian Goodfellow, Tatiane C. Nogueira, Erik Verbeken, Jelle Matthijnssens, Peter de Witte, Johan Neyts, Joana Rocha-Pereira. A robust human norovirus replication model in zebrafish larvae. PLOS Pathogens. 2019; 15 (9):e1008009.
Chicago/Turabian StyleJana Van Dycke; Annelii Ny; Nádia Conceição-Neto; Jan Maes; Myra Hosmillo; Arno Cuvry; Ian Goodfellow; Tatiane C. Nogueira; Erik Verbeken; Jelle Matthijnssens; Peter de Witte; Johan Neyts; Joana Rocha-Pereira. 2019. "A robust human norovirus replication model in zebrafish larvae." PLOS Pathogens 15, no. 9: e1008009.
Viral gastroenteritis is an important cause of morbidity and mortality worldwide, being particularly severe for children under the age of five. The most common viral agents of gastroenteritis are noroviruses, rotaviruses, sapoviruses, astroviruses and adenoviruses, however, no specific antiviral treatment exists today against any of these pathogens. We here discuss the feasibility of developing a broad-spectrum antiviral treatment against these diarrhea-causing viruses. This review focuses on the viral polymerase as an antiviral target, as this is the most conserved viral protein among the diverse viral families to which these viruses belong to. We describe the functional and structural similarities of the different viral polymerases, the antiviral effect of reported polymerase inhibitors and highlight common features that might be exploited in an attempt of designing such pan-polymerase inhibitor.
Marcella Bassetto; Jana Van Dycke; Johan Neyts; Andrea Brancale; Joana Rocha-Pereira. Targeting the Viral Polymerase of Diarrhea-Causing Viruses as a Strategy to Develop a Single Broad-Spectrum Antiviral Therapy. Viruses 2019, 11, 173 .
AMA StyleMarcella Bassetto, Jana Van Dycke, Johan Neyts, Andrea Brancale, Joana Rocha-Pereira. Targeting the Viral Polymerase of Diarrhea-Causing Viruses as a Strategy to Develop a Single Broad-Spectrum Antiviral Therapy. Viruses. 2019; 11 (2):173.
Chicago/Turabian StyleMarcella Bassetto; Jana Van Dycke; Johan Neyts; Andrea Brancale; Joana Rocha-Pereira. 2019. "Targeting the Viral Polymerase of Diarrhea-Causing Viruses as a Strategy to Develop a Single Broad-Spectrum Antiviral Therapy." Viruses 11, no. 2: 173.
The order of Bunyavirales includes numerous (re)emerging viruses that collectively have a major impact on human and animal health worldwide. There are no vaccines for human use or antiviral drugs available to prevent or treat infections with any of these viruses. The development of efficacious and safe drugs and vaccines is a pressing matter. Ideally, such antivirals possess pan‐bunyavirus antiviral activity, allowing the containment of every bunya‐related threat. The fact that many bunyaviruses need to be handled in laboratories with biosafety level 3 or 4, the great variety of species and the frequent emergence of novel species complicate such efforts. We here examined the potential druggable targets of bunyaviruses, together with the level of conservation of their biological functions, structure, and genetic similarity by means of heatmap analysis. In the light of this, we revised the available models and tools currently available, pointing out directions for antiviral drug discovery.
Sebastiaan Ter Horst; Nádia Conceição-Neto; Johan Neyts; Joana Rocha-Pereira. Structural and functional similarities in bunyaviruses: Perspectives for pan-bunya antivirals. Reviews in Medical Virology 2019, 29, e2039 .
AMA StyleSebastiaan Ter Horst, Nádia Conceição-Neto, Johan Neyts, Joana Rocha-Pereira. Structural and functional similarities in bunyaviruses: Perspectives for pan-bunya antivirals. Reviews in Medical Virology. 2019; 29 (3):e2039.
Chicago/Turabian StyleSebastiaan Ter Horst; Nádia Conceição-Neto; Johan Neyts; Joana Rocha-Pereira. 2019. "Structural and functional similarities in bunyaviruses: Perspectives for pan-bunya antivirals." Reviews in Medical Virology 29, no. 3: e2039.
Human noroviruses (HuNoVs) are an important cause of epidemic and endemic acute gastroenteritis worldwide; annually about 700 million people develop a HuNoV infection resulting in ∼219,000 deaths and a societal cost estimated at 60 billion US dollars 1. The lack of robust small animal models has significantly hindered the understanding of norovirus biology and the development of effective therapeutics against HuNoV. Here we report that HuNoV GI and GII replicate to high titers in zebrafish (Danio rerio) larvae; replication peaks at day 2 post infection and is detectable for at least 6 days. HuNoV is detected in cells of the hematopoietic lineage, the intestine, liver and pancreas. Antiviral treatment reduces HuNoV replication by >2 log10, showing that this model is suited for antiviral studies. Downregulation of fucosyltransferase 8 (fut8) in the larvae reduces HuNoV replication, highlighting a common feature with infection in humans. Zebrafish larvae constitute a simple and robust replication model that will largely facilitate studies of HuNoV biology and the development of antiviral strategies.
Jana Van Dycke; Annelii Ny; Nadia Conceicao-Neto; Jan Maes; Myra Hosmillo; Arno Cuvry; Ian Goodfellow; Tatiane C De Araujo Nogueira; Erik Verbeken; Jelle Matthijnssens; Peter De Witte; Johan Neyts; Joana Rocha-Pereira. A robust human norovirus replication model in zebrafish larvae. 2019, 1 .
AMA StyleJana Van Dycke, Annelii Ny, Nadia Conceicao-Neto, Jan Maes, Myra Hosmillo, Arno Cuvry, Ian Goodfellow, Tatiane C De Araujo Nogueira, Erik Verbeken, Jelle Matthijnssens, Peter De Witte, Johan Neyts, Joana Rocha-Pereira. A robust human norovirus replication model in zebrafish larvae. . 2019; ():1.
Chicago/Turabian StyleJana Van Dycke; Annelii Ny; Nadia Conceicao-Neto; Jan Maes; Myra Hosmillo; Arno Cuvry; Ian Goodfellow; Tatiane C De Araujo Nogueira; Erik Verbeken; Jelle Matthijnssens; Peter De Witte; Johan Neyts; Joana Rocha-Pereira. 2019. "A robust human norovirus replication model in zebrafish larvae." , no. : 1.
The type III interferon (IFN-λ) family includes 4 IFN-λ subtypes in man. In the mouse, only the genes coding for IFN-λ2 and -λ3 are present. Unlike mouse and human type I IFNs (IFN-α/β), which exhibit strong species specificity, type III IFNs were reported to act in a cross-specific manner. We reexamined the cross-specificity and observed that mouse and human IFN-λ exhibit some species specificity, although much less than type I IFNs. Mouse IFN-λ3 displayed clear species specificity, being 25-fold less active in human cells than the closely related mouse IFN-λ2. This specificity likely depends on amino acids in α helices A and F that diverged from other IFN-λ sequences. Human IFN-λ4, in contrast, retained high activity in mouse cells. We next developed a firefly luciferase-based reporter cell line, named Fawa-λ-luc, to detect IFN-λ in biological fluids with high specificity and sensitivity. Fawa-λ-luc cells, derived from mouse epithelial cells that are responsive to IFN-λ, were made nonresponsive to type I IFNs by inactivation of the Ifnar2 gene and strongly responsive to IFN-λ by overexpression of the mouse IFNLR1. This bioassay was as sensitive as a commercially available enzyme-linked immunosorbent assay in detecting mouse IFN-λ in cell culture supernatant, as well as in serum and bronchoalveolar lavage samples of virus-infected mice. The assay also enabled the sensitive detection of human IFN-λ activity, including that of the divergent IFN-λ4 with a bias, however, due to variable activity of IFN-λ subtypes.
Sophie Jacobs; Fanny Wavreil; Bert Schepens; Hans Henrik Gad; Rune Hartmann; Joana Rocha-Pereira; Johan Neyts; Xavier Saelens; Thomas Michiels. Species Specificity of Type III Interferon Activity and Development of a Sensitive Luciferase-Based Bioassay for Quantitation of Mouse Interferon-λ. Journal of Interferon & Cytokine Research 2018, 38, 469 -479.
AMA StyleSophie Jacobs, Fanny Wavreil, Bert Schepens, Hans Henrik Gad, Rune Hartmann, Joana Rocha-Pereira, Johan Neyts, Xavier Saelens, Thomas Michiels. Species Specificity of Type III Interferon Activity and Development of a Sensitive Luciferase-Based Bioassay for Quantitation of Mouse Interferon-λ. Journal of Interferon & Cytokine Research. 2018; 38 (11):469-479.
Chicago/Turabian StyleSophie Jacobs; Fanny Wavreil; Bert Schepens; Hans Henrik Gad; Rune Hartmann; Joana Rocha-Pereira; Johan Neyts; Xavier Saelens; Thomas Michiels. 2018. "Species Specificity of Type III Interferon Activity and Development of a Sensitive Luciferase-Based Bioassay for Quantitation of Mouse Interferon-λ." Journal of Interferon & Cytokine Research 38, no. 11: 469-479.
A safe and highly efficient antiviral is needed for the prophylaxis and/or treatment of viral diarrhea. We here demonstrate the in vitro antiviral activity of four 2'-C-methyl nucleoside analogues against noro-, rota-, and sapoviruses. The most potent nucleoside analogue, 7-deaza-2'-C-methyladenosine, inhibits replication of these viruses with a 50% effective concentration < 5 µM. Mechanistically, we demonstrate that the 2'-C-methyl nucleoside analogues act by inhibiting transcription of the rotavirus genome. This provides the first evidence that a single viral-diarrhea-targeted treatment can be developed through a viral-polymerase-targeting small molecule.
Jana Van Dycke; Francesca Arnoldi; Guido Papa; Justine Vandepoele; Oscar R Burrone; Eloise Mastrangelo; Delia Tarantino; Elisabeth Heylen; Johan Neyts; Joana Rocha-Pereira. A Single Nucleoside Viral Polymerase Inhibitor Against Norovirus, Rotavirus, and Sapovirus-Induced Diarrhea. Journal of Infectious Diseases 2018, 218, 1753 -1758.
AMA StyleJana Van Dycke, Francesca Arnoldi, Guido Papa, Justine Vandepoele, Oscar R Burrone, Eloise Mastrangelo, Delia Tarantino, Elisabeth Heylen, Johan Neyts, Joana Rocha-Pereira. A Single Nucleoside Viral Polymerase Inhibitor Against Norovirus, Rotavirus, and Sapovirus-Induced Diarrhea. Journal of Infectious Diseases. 2018; 218 (11):1753-1758.
Chicago/Turabian StyleJana Van Dycke; Francesca Arnoldi; Guido Papa; Justine Vandepoele; Oscar R Burrone; Eloise Mastrangelo; Delia Tarantino; Elisabeth Heylen; Johan Neyts; Joana Rocha-Pereira. 2018. "A Single Nucleoside Viral Polymerase Inhibitor Against Norovirus, Rotavirus, and Sapovirus-Induced Diarrhea." Journal of Infectious Diseases 218, no. 11: 1753-1758.
Bio-protocol is an online peer-reviewed protocol journal. Its mission is to make life science research more efficient and reproducible by curating and hosting high quality, free access protocols.
Jana Dycke; Johan Neyts; Joana Rocha-Pereira. Assessing the Efficacy of Small Molecule Inhibitors in a Mouse Model of Persistent Norovirus Infection. BIO-PROTOCOL 2018, 8, 1 .
AMA StyleJana Dycke, Johan Neyts, Joana Rocha-Pereira. Assessing the Efficacy of Small Molecule Inhibitors in a Mouse Model of Persistent Norovirus Infection. BIO-PROTOCOL. 2018; 8 (9):1.
Chicago/Turabian StyleJana Dycke; Johan Neyts; Joana Rocha-Pereira. 2018. "Assessing the Efficacy of Small Molecule Inhibitors in a Mouse Model of Persistent Norovirus Infection." BIO-PROTOCOL 8, no. 9: 1.
Human noroviruses are highly efficient in person to person transmission thus associated with explosive outbreaks of acute gastroenteritis. Outbreak control is limited to disinfection and isolation measures. Strategies to control the spread of noroviruses should be developed and models to study norovirus transmission will greatly facilitate this. Here, a mouse-to-mouse transmission model, in which mice develop acute murine norovirus (MNV)-induced diarrhea, was used to explore the role of interferon lambda (IFN-λ) in the control of a norovirus infection. Sentinel AG129 mice [deficient in IFN-α/β and IFN-γ receptors] that were co-housed with MNV-infected mice shedding high amounts of virus in their stool, developed a MNV-infection with associated diarrhea. Inoculation of such sentinel mice with an IFN-λ expression plasmid resulted in the production of circulating IFN-λ and upregulation of the expression of IFN-stimulated genes (ISGs) of the gut. Injection of the IFN-λ-expressing plasmid to sentinels prevents MNV-induced disease upon exposure to MNV-infected mice, as well as MNV replication in the small intestine, the associated signs of inflammation and the mounting of a specific IgG-based immune response. This demonstrates that IFN-λ can alone mediate protection against transmission of norovirus. The development of a simple delivery method for IFN-λ could be explored as a strategy to control norovirus outbreaks and protect vulnerable populations such as the elderly and immunocompromised.
Joana Rocha-Pereira; Sophie Jacobs; Sam Noppen; Eric Verbeken; Thomas Michiels; Johan Neyts. Interferon lambda (IFN-λ) efficiently blocks norovirus transmission in a mouse model. Antiviral Research 2018, 149, 7 -15.
AMA StyleJoana Rocha-Pereira, Sophie Jacobs, Sam Noppen, Eric Verbeken, Thomas Michiels, Johan Neyts. Interferon lambda (IFN-λ) efficiently blocks norovirus transmission in a mouse model. Antiviral Research. 2018; 149 ():7-15.
Chicago/Turabian StyleJoana Rocha-Pereira; Sophie Jacobs; Sam Noppen; Eric Verbeken; Thomas Michiels; Johan Neyts. 2018. "Interferon lambda (IFN-λ) efficiently blocks norovirus transmission in a mouse model." Antiviral Research 149, no. : 7-15.
A series of 4-substituted 3,4-dihydropyrimidine-2-ones (DHPM) was synthesized, characterized by IR, 1H NMR, 13C NMR and HRMS spectra. The compounds were evaluated in vitro for their antiviral activity against a broad range of DNA and RNA viruses, along with assessment for potential cytotoxicity in diverse mammalian cell lines. Compound 4m, which possesses a long lipophilic side chain, was found to be a potent and selective inhibitor of Punta Toro virus, a member of the Bunyaviridae. For Rift Valley fever virus, which is another Bunyavirus, the activity of 4m was negligible. DHPMs with a C-4 aryl moiety bearing halogen substitution (4b, 4c and 4d) were found to be cytotoxic in MT4 cells.
Dhanabal Kumarasamy; Biswajit Gopal Roy; Joana Rocha-Pereira; Johan Neyts; SatheeshKumar Nanjappan; Subhasis Maity; Musfiqua Mookerjee; Lieve Naesens. Synthesis and in vitro antiviral evaluation of 4-substituted 3,4-dihydropyrimidinones. Bioorganic & Medicinal Chemistry Letters 2016, 27, 139 -142.
AMA StyleDhanabal Kumarasamy, Biswajit Gopal Roy, Joana Rocha-Pereira, Johan Neyts, SatheeshKumar Nanjappan, Subhasis Maity, Musfiqua Mookerjee, Lieve Naesens. Synthesis and in vitro antiviral evaluation of 4-substituted 3,4-dihydropyrimidinones. Bioorganic & Medicinal Chemistry Letters. 2016; 27 (2):139-142.
Chicago/Turabian StyleDhanabal Kumarasamy; Biswajit Gopal Roy; Joana Rocha-Pereira; Johan Neyts; SatheeshKumar Nanjappan; Subhasis Maity; Musfiqua Mookerjee; Lieve Naesens. 2016. "Synthesis and in vitro antiviral evaluation of 4-substituted 3,4-dihydropyrimidinones." Bioorganic & Medicinal Chemistry Letters 27, no. 2: 139-142.
Human noroviruses are the leading cause of foodborne illness causing both acute and chronic gastroenteritis. In recent years, a number of vaccine candidates entered (pre-) clinical development and the first efforts to develop antiviral therapy have been made. We here discuss aspects of norovirus genetic evolution, persistence in immunocompromised patients as well as the risk and potential consequences of resistance development toward future antiviral drugs.
Joana Rocha-Pereira; Jana Van Dycke; Johan Neyts. Norovirus genetic diversity and evolution: implications for antiviral therapy. Current Opinion in Virology 2016, 20, 92 -98.
AMA StyleJoana Rocha-Pereira, Jana Van Dycke, Johan Neyts. Norovirus genetic diversity and evolution: implications for antiviral therapy. Current Opinion in Virology. 2016; 20 ():92-98.
Chicago/Turabian StyleJoana Rocha-Pereira; Jana Van Dycke; Johan Neyts. 2016. "Norovirus genetic diversity and evolution: implications for antiviral therapy." Current Opinion in Virology 20, no. : 92-98.