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Article: Current concepts in the development of therapeutics against human and animal coronavirus diseases by targeting NP.

Tseng, Yeu-Yang / Liao, Guan-Ru / Lien, Abigail / Hsu, Wei-Li

Computational and structural biotechnology journal

2021 Volume 19, Page(s) 1072–1080

Abstract: The coronavirus (CoV) infects a broad range of hosts including humans as well as a variety of animals. It has gained overwhelming concerns since the emergence of deadly human coronaviruses (HCoVs), severe acute respiratory syndrome coronavirus (SARS-CoV) ...

Abstract	The coronavirus (CoV) infects a broad range of hosts including humans as well as a variety of animals. It has gained overwhelming concerns since the emergence of deadly human coronaviruses (HCoVs), severe acute respiratory syndrome coronavirus (SARS-CoV) in 2003, followed by Middle East respiratory syndrome coronavirus (MERS-CoV) in 2015. Very recently, special attention has been paid to the novel coronavirus disease 2019 (COVID-19) caused by SARS-CoV-2 due to its high mobility and mortality. As the COVID-19 pandemic continues, despite vast research efforts, the effective pharmaceutical interventions are still not available for clinical uses. Both expanded knowledge on structure insights and the essential function of viral nucleocapsid (N) protein are key basis for the development of novel, and potentially, a broad-spectrum inhibitor against coronavirus diseases. This review aimed to delineate the current research from the perspective of biochemical and structural study in cell-based assays as well as virtual screen approaches to identify N protein antagonists targeting not only HCoVs but also animal CoVs.
Language	English
Publishing date	2021-01-30
Publishing country	Netherlands
Document type	Journal Article ; Review
ISSN	2001-0370
ISSN	2001-0370
DOI	10.1016/j.csbj.2021.01.032
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Article ; Online: Characterization of influenza B viruses with reduced susceptibility to influenza neuraminidase inhibitors.

Brown, Sook Kwan / Tseng, Yeu-Yang / Aziz, Ammar / Baz, Mariana / Barr, Ian G

Antiviral research

2022 Volume 200, Page(s) 105280

Abstract: A total of 3425 influenza B viruses collected from the Asia-Pacific region were tested against the four registered neuraminidase inhibitors (NAIs) (oseltamivir carboxylate, zanamivir, peramivir and laninamivir) as part of the routine surveillance work at ...

Abstract	A total of 3425 influenza B viruses collected from the Asia-Pacific region were tested against the four registered neuraminidase inhibitors (NAIs) (oseltamivir carboxylate, zanamivir, peramivir and laninamivir) as part of the routine surveillance work at the WHO Collaborating Centre for Research and Reference on Influenza, Melbourne between 2016 and 2020. Forty-five influenza B viruses with reduced susceptibility to one or more NAIs were identified. While the majority of these had neuraminidase (NA) mutations that were known to confer NAIs resistance, fifteen had NA mutations that had not been confirmed as being responsible for reduced NAIs susceptibility. Eleven of these NA mutations of concern were investigated using reverse genetics (RG) techniques to verify that these mutations were the cause of the reduced NAI susceptibility. All mutations were introduced separately into the NA of B/Brisbane/27/2016 (a B Victoria-lineage virus) or B/Yamanashi/166/98 (a B Yamagata-lineage virus) and the effects of these were analysed by an in vitro NAI assay. The T146K substitution in the NA of B Victoria and Yamagata-lineages resulted in a large increase in the IC
MeSH term(s)	Antiviral Agents/pharmacology ; Antiviral Agents/therapeutic use ; Drug Resistance, Viral/genetics ; Enzyme Inhibitors/pharmacology ; Enzyme Inhibitors/therapeutic use ; Humans ; Influenza B virus/genetics ; Influenza, Human/drug therapy ; Neuraminidase/genetics ; Neuraminidase/therapeutic use ; Oseltamivir/pharmacology ; Oseltamivir/therapeutic use ; Zanamivir/pharmacology ; Zanamivir/therapeutic use
Chemical Substances	Antiviral Agents ; Enzyme Inhibitors ; Oseltamivir (20O93L6F9H) ; Neuraminidase (EC 3.2.1.18) ; Zanamivir (L6O3XI777I)
Language	English
Publishing date	2022-03-15
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	306628-9
ISSN	1872-9096 ; 0166-3542
ISSN (online)	1872-9096
ISSN	0166-3542
DOI	10.1016/j.antiviral.2022.105280
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Article ; Online: The orf virus (ORFV) protein OV20.0 interacts with the microprocessor complex subunit DGCR8 to regulate miRNA biogenesis and ORFV infection.

Liao, Guan-Ru / Tseng, Yeu-Yang / Tseng, Ching-Yu / Lo, Chen-Yu / Hsu, Wei-Li

FEBS letters

2021 Volume 595, Issue 23, Page(s) 2897–2908

Abstract: Cellular double-stranded RNA-binding proteins (DRBPs) play important roles in the regulation of innate immune responses and microRNA (miRNA) biogenesis. The current study aimed to understand whether OV20.0, a DRBP of orf virus (ORFV), is involved in ... ...

Abstract	Cellular double-stranded RNA-binding proteins (DRBPs) play important roles in the regulation of innate immune responses and microRNA (miRNA) biogenesis. The current study aimed to understand whether OV20.0, a DRBP of orf virus (ORFV), is involved in cellular RNA biogenesis via association with host DRBPs. We found that OV20.0 interacts with DiGeorge syndrome critical region 8 (DGCR8), a subunit of the miRNA processor complex, and binds to primary- and precursor-miRNA. Additionally, OV20.0 regulates DGCR8 expression in multiple ways, including through interaction with the DGCR8 protein and binding to DGCR8 mRNA. Lastly, our data show that DGCR8 plays an antiviral role against ORFV infection, whereas it is beneficial for influenza virus propagation, indicating that the underlying mechanisms could be diverse among different viruses.
MeSH term(s)	A549 Cells ; Animals ; Dogs ; Ecthyma, Contagious/metabolism ; Ecthyma, Contagious/virology ; HEK293 Cells ; Humans ; Madin Darby Canine Kidney Cells ; MicroRNAs/genetics ; MicroRNAs/metabolism ; Orf virus/pathogenicity ; Protein Binding ; RNA, Messenger/metabolism ; RNA-Binding Proteins/metabolism ; Viral Proteins/metabolism
Chemical Substances	DGCR8 protein, human ; MicroRNAs ; OV20.0L protein, Orf virus ; RNA, Messenger ; RNA-Binding Proteins ; Viral Proteins
Language	English
Publishing date	2021-11-22
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	212746-5
ISSN	1873-3468 ; 0014-5793
ISSN (online)	1873-3468
ISSN	0014-5793
DOI	10.1002/1873-3468.14231
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Article ; Online: Deletion of gene OV132 attenuates Orf virus more effectively than gene OV112

Yamada, Yumiko / Chuang, Shih-Te / Tseng, Ching-Yu / Liao, Guan-Ru / Liu, Shin-Wu / Tseng, Yeu-Yang / Lin, Fong-Yuan / Xu, Weili

Appl Microbiol Biotechnol. 2023 Feb., v. 107, no. 2-3 p.835-851

2023

Abstract: Orf virus (ORFV), a Parapoxvirus in Poxviridae, infects sheep and goats resulting in contagious pustular dermatitis. ORFV is regarded as a promising viral vector candidate for vaccine development and oncolytic virotherapy. Owing to their potential ... ...

Abstract	Orf virus (ORFV), a Parapoxvirus in Poxviridae, infects sheep and goats resulting in contagious pustular dermatitis. ORFV is regarded as a promising viral vector candidate for vaccine development and oncolytic virotherapy. Owing to their potential clinical application, safety concerns have become increasingly important. Deletion of either the OV132 (encoding vascular endothelial growth factor, VEGF) or OV112 (encoding the chemokine binding protein, CBP) genes reduced ORFV infectivity, which has been independently demonstrated in the NZ2 and NZ7 strains, respectively. This study revealed that the VEGF and CBP gene sequences of the local strain (TW/Hoping) shared a similarity of 47.01% with NZ2 and 90.56% with NZ7. Due to the high sequence divergence of these two immunoregulatory genes among orf viral strains, their contribution to the pathogenicity of Taiwanese ORFV isolates was comparatively characterized. Initially, two ORFV recombinants were generated, in which either the VEGF or CBP gene was deleted and replaced with the reporter gene EGFP. In vitro assays indicated that both the VEGF-deletion mutant ORFV-VEGFΔ-EGFP and the CBP deletion mutant ORFV-CBPΔ-EGFP were attenuated in cells. In particular, ORFV-VEGFΔ-EGFP significantly reduced plaque size and virus yield compared to ORFV-CBPΔ-EGFP and the wild-type control. Similarly, in vivo analysis revealed no virus yield in the goat skin biopsy infected by ORFV-VEGFΔ-EGFP, and significantly reduced the virus yield of ORFV-CBPΔ-EGFP relative to the wild-type control. These results confirmed the loss of virulence of both deletion mutants in the Hoping strain, whereas the VEGF-deletion mutant was more attenuated than the CBP deletion strain in both cell and goat models. KEY POINTS: • VEGF and CBP genes are crucial in ORFV pathogenesis in the TW/Hoping strain • The VEGF-deletion mutant virus was severely attenuated in both cell culture and animal models • Deletion mutant viruses are advantageous vectors for the development of vaccines and therapeutic regimens
Keywords	Orf virus ; biopsy ; cell culture ; chemokines ; contagious ecthyma ; goats ; immunomodulation ; mutants ; pathogenesis ; reporter genes ; sheep ; vaccine development ; vascular endothelial growth factors ; virulence ; viruses
Language	English
Dates of publication	2023-02
Size	p. 835-851.
Publishing place	Springer Berlin Heidelberg
Document type	Article ; Online
ZDB-ID	392453-1
ISSN	1432-0614 ; 0171-1741 ; 0175-7598
ISSN (online)	1432-0614
ISSN	0171-1741 ; 0175-7598
DOI	10.1007/s00253-022-12323-0
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Article ; Online: Deletion of gene OV132 attenuates Orf virus more effectively than gene OV112.

Yamada, Yumiko / Chuang, Shih-Te / Tseng, Ching-Yu / Liao, Guan-Ru / Liu, Shin-Wu / Tseng, Yeu-Yang / Lin, Fong-Yuan / Hsu, Wei-Li

Applied microbiology and biotechnology

2022 Volume 107, Issue 2-3, Page(s) 835–851

Abstract	Orf virus (ORFV), a Parapoxvirus in Poxviridae, infects sheep and goats resulting in contagious pustular dermatitis. ORFV is regarded as a promising viral vector candidate for vaccine development and oncolytic virotherapy. Owing to their potential clinical application, safety concerns have become increasingly important. Deletion of either the OV132 (encoding vascular endothelial growth factor, VEGF) or OV112 (encoding the chemokine binding protein, CBP) genes reduced ORFV infectivity, which has been independently demonstrated in the NZ2 and NZ7 strains, respectively. This study revealed that the VEGF and CBP gene sequences of the local strain (TW/Hoping) shared a similarity of 47.01% with NZ2 and 90.56% with NZ7. Due to the high sequence divergence of these two immunoregulatory genes among orf viral strains, their contribution to the pathogenicity of Taiwanese ORFV isolates was comparatively characterized. Initially, two ORFV recombinants were generated, in which either the VEGF or CBP gene was deleted and replaced with the reporter gene EGFP. In vitro assays indicated that both the VEGF-deletion mutant ORFV-VEGFΔ-EGFP and the CBP deletion mutant ORFV-CBPΔ-EGFP were attenuated in cells. In particular, ORFV-VEGFΔ-EGFP significantly reduced plaque size and virus yield compared to ORFV-CBPΔ-EGFP and the wild-type control. Similarly, in vivo analysis revealed no virus yield in the goat skin biopsy infected by ORFV-VEGFΔ-EGFP, and significantly reduced the virus yield of ORFV-CBPΔ-EGFP relative to the wild-type control. These results confirmed the loss of virulence of both deletion mutants in the Hoping strain, whereas the VEGF-deletion mutant was more attenuated than the CBP deletion strain in both cell and goat models. KEY POINTS: • VEGF and CBP genes are crucial in ORFV pathogenesis in the TW/Hoping strain • The VEGF-deletion mutant virus was severely attenuated in both cell culture and animal models • Deletion mutant viruses are advantageous vectors for the development of vaccines and therapeutic regimens.
MeSH term(s)	Animals ; Ecthyma, Contagious/pathology ; Goats ; Orf virus/genetics ; Sheep ; Skin ; Vascular Endothelial Growth Factor A/genetics ; Genes, Viral
Chemical Substances	Vascular Endothelial Growth Factor A
Language	English
Publishing date	2022-12-09
Publishing country	Germany
Document type	Journal Article
ZDB-ID	392453-1
ISSN	1432-0614 ; 0171-1741 ; 0175-7598
ISSN (online)	1432-0614
ISSN	0171-1741 ; 0175-7598
DOI	10.1007/s00253-022-12323-0
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Zs.A 2229: Show issues

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Article ; Online: A variant NS1 protein from H5N2 avian influenza virus suppresses PKR activation and promotes replication and virulence in mammals.

Chung, Yun-Ting / Kuan, Chih-Ying / Liao, Guan-Ru / Albrecht, Randy A / Tseng, Yeu-Yang / Hsu, Yu-Chen / Ou, Shan-Chia / Hsu, Wei-Li

Emerging microbes & infections

2022 Volume 11, Issue 1, Page(s) 2291–2303

Abstract: Highly pathogenic avian influenza viruses (HPAIVs) frequently receive global attention as threats to public health. The NS1 protein is a key virulence factor known to impair host antiviral responses. The study herein revealed HPAIV H5N2 NS gene encoded ... ...

Abstract	Highly pathogenic avian influenza viruses (HPAIVs) frequently receive global attention as threats to public health. The NS1 protein is a key virulence factor known to impair host antiviral responses. The study herein revealed HPAIV H5N2 NS gene encoded additional protein; a truncated NS1 variant, designated NS3, produced by alternative splicing of the NS transcript. To examine the function of NS3 during infection, we generated recombinant viruses expressing either full-length NS1 (RG-AIV-T375G) or NS3 (RG-AIV-NS3). Interestingly, RG-AIV-NS3 virus produced higher titres than RG-AIV-T375G in multiple mammalian cell lines. However, RG-AIV-T375G exhibited a replication advantage over RG-AIV-NS3 in chicken DF-1 cells, indicating that host cell identity dictates the effect of NS3 on viral replication. In mice and mammalian cells, RG-AIV-NS3 infection elicited higher level of cytokines, including IFN-β, MX and TNF-α, potentially due to its higher replication activity. Based on mini-genome assay, NS3 had pronounced effects on viral replication machinery. Surprisingly, NS3 retained an interaction with PKR and suppressed PKR activation despite its lack of amino-acid residues 126-167. The poor replication ability of RG-AIV-T375G was partially restored in cells deficient in PKR suggesting that full-length NS1 may be insufficient to suppress PKR function. Notably, virulence of the full-length NS1-expressing RG-AIV-T375G virus was highly attenuated in mice when compared to RG-AIV-NS3. In summary, our study reveals the existence and function of a previously unidentified H5N2 viral protein, NS3. We found that NS3 is functionally distinct from NS1 protein, as it enhances viral replication and pathogenicity in mammalian systems, potentially via suppression of PKR activity.
MeSH term(s)	Animals ; Antiviral Agents ; Birds/virology ; Influenza A Virus, H5N2 Subtype ; Influenza in Birds ; Mammals ; Mice ; Receptors, G-Protein-Coupled ; Tumor Necrosis Factor-alpha ; Viral Nonstructural Proteins/metabolism ; Virulence/genetics ; Virulence Factors/genetics ; Virus Replication/genetics
Chemical Substances	Antiviral Agents ; PKR1 protein, mouse ; Receptors, G-Protein-Coupled ; Tumor Necrosis Factor-alpha ; Viral Nonstructural Proteins ; Virulence Factors
Language	English
Publishing date	2022-08-18
Publishing country	United States
Document type	Journal Article
ZDB-ID	2681359-2
ISSN	2222-1751 ; 2222-1751
ISSN (online)	2222-1751
ISSN	2222-1751
DOI	10.1080/22221751.2022.2114853
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Article: The orf virus (ORFV) protein OV20.0 interacts with the microprocessor complex subunit DGCR8 to regulate miRNA biogenesis and ORFV infection

Liao, Guan‐Ru / Tseng, Yeu‐Yang / Tseng, Ching‐Yu / Lo, Chen‐Yu / Hsu, Wei‐Li

FEBS letters. 2021 Dec., v. 595, no. 23

2021

Abstract: Cellular double‐stranded RNA‐binding proteins (DRBPs) play important roles in the regulation of innate immune responses and microRNA (miRNA) biogenesis. The current study aimed to understand whether OV20.0, a DRBP of orf virus (ORFV), is involved in ... ...

Abstract	Cellular double‐stranded RNA‐binding proteins (DRBPs) play important roles in the regulation of innate immune responses and microRNA (miRNA) biogenesis. The current study aimed to understand whether OV20.0, a DRBP of orf virus (ORFV), is involved in cellular RNA biogenesis via association with host DRBPs. We found that OV20.0 interacts with DiGeorge syndrome critical region 8 (DGCR8), a subunit of the miRNA processor complex, and binds to primary‐ and precursor‐miRNA. Additionally, OV20.0 regulates DGCR8 expression in multiple ways, including through interaction with the DGCR8 protein and binding to DGCR8 mRNA. Lastly, our data show that DGCR8 plays an antiviral role against ORFV infection, whereas it is beneficial for influenza virus propagation, indicating that the underlying mechanisms could be diverse among different viruses.
Keywords	Orf virus ; Orthomyxoviridae ; biogenesis ; microRNA
Language	English
Dates of publication	2021-12
Size	p. 2897-2908.
Publishing place	John Wiley & Sons, Ltd
Document type	Article
Note	JOURNAL ARTICLE
ZDB-ID	212746-5
ISSN	1873-3468 ; 0014-5793
ISSN (online)	1873-3468
ISSN	0014-5793
DOI	10.1002/1873-3468.14231
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Article ; Online: K160 in the RNA-binding domain of the orf virus virulence factor OV20.0 is critical for its functions in counteracting host antiviral defense.

Liao, Guan-Ru / Tseng, Yeu-Yang / Tseng, Ching-Yu / Huang, Ying-Ping / Tsai, Ching-Hsiu / Liu, Hao-Ping / Hsu, Wei-Li

FEBS letters

2021 Volume 595, Issue 12, Page(s) 1721–1733

Abstract: The OV20.0 virulence factor of orf virus antagonizes host antiviral responses. One mechanism through which it functions is by inhibiting activation of the dsRNA-activated protein kinase R (PKR) by sequestering dsRNA and by physically interacting with PKR. ...

Abstract	The OV20.0 virulence factor of orf virus antagonizes host antiviral responses. One mechanism through which it functions is by inhibiting activation of the dsRNA-activated protein kinase R (PKR) by sequestering dsRNA and by physically interacting with PKR. Sequence alignment indicated that several key residues critical for dsRNA binding were conserved in OV20.0, and their contribution to OV20.O function was investigated in this study. We found that residues F141, K160, and R164 were responsible for the dsRNA-binding ability of OV20.0. Interestingly, mutation at K160 (K160A) diminished the OV20.0-PKR interaction and further reduced the inhibitory effect of OV20.0 on PKR activation. Nevertheless, OV20.0 homodimerization was not influenced by K160A. The contribution of the dsRNA-binding domain and K160 to the suppression of RNA interference by OV20.0 was further demonstrated in plants. In summary, K160 is essential for the function of OV20.0, particularly its interaction with dsRNA and PKR that ultimately contributes to the suppression of PKR activation.
MeSH term(s)	HEK293 Cells ; Humans ; Orf virus/genetics ; Orf virus/metabolism ; Orf virus/pathogenicity ; Protein Domains ; RNA, Double-Stranded/genetics ; RNA, Double-Stranded/metabolism ; Viral Proteins/genetics ; Viral Proteins/metabolism ; Virulence Factors/genetics ; Virulence Factors/metabolism ; eIF-2 Kinase/genetics ; eIF-2 Kinase/metabolism
Chemical Substances	OV20.0L protein, Orf virus ; RNA, Double-Stranded ; Viral Proteins ; Virulence Factors ; EIF2AK2 protein, human (EC 2.7.11.1) ; eIF-2 Kinase (EC 2.7.11.1)
Language	English
Publishing date	2021-05-27
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	212746-5
ISSN	1873-3468 ; 0014-5793
ISSN (online)	1873-3468
ISSN	0014-5793
DOI	10.1002/1873-3468.14099
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Article ; Online: Establishment and characterization of transformed goat primary cells by expression of simian virus 40 large T antigen for orf virus propagations.

Yumiko Yamada / Guan-Ru Liao / Ching-Yu Tseng / Yeu-Yang Tseng / Wei-Li Hsu

PLoS ONE, Vol 14, Iss 12, p e

2019 Volume 0226105

Abstract: Due to the limited host range of orf virus (ORFV), primary cells derived from its natural hosts, such as goats and sheep, are recommended for isolation and propagation of wild type ORFV. This situation limits the option for the study of virus-host ... ...

Abstract	Due to the limited host range of orf virus (ORFV), primary cells derived from its natural hosts, such as goats and sheep, are recommended for isolation and propagation of wild type ORFV. This situation limits the option for the study of virus-host interaction during ORFV infection since primary cells only support a few numbers of passages. SV40 T antigen is a viral oncoprotein that can abrogate replicative senescence, leading to an extended life span of cells. In this study, the transformation of two goat primary cells, fibroblast (FB) and testis (GT) cells, were achieved by stably expressing SV40 T antigen using the lentiviral technique. The presence of the gene encoding SV40 T antigen was validated by polymerase chain reaction (PCR) and western blot analyses. As evidenced by immunofluorescent microscopy, the two types of cells expressing SV40 T antigen (namely, FBT and GTT) were purified to homogeneity. Moreover, faster growth kinetics and a lower serum dependency were noticed in FBT and GTT, as compared with their counterpart parental cells. FBT and GTT remain permissive and can form plaque of ORFV, despite with different profiles; generally speaking, with SV40 T expression, ORFV forms plaques with smaller size and distinct margin. Most importantly, the prolonged life span of goat FBT and GTT serves as an ideal cell culture resource for ORFV isolation from the field, studies of ORFV pathogenesis and efficient vaccine development.
Keywords	Medicine ; R ; Science ; Q
Subject code	570
Language	English
Publishing date	2019-01-01T00:00:00Z
Publisher	Public Library of Science (PLoS)
Document type	Article ; Online
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Article ; Online: Increasing antigen presentation on HSV-1-infected cells increases lesion size but does not alter neural infection or latency.

Russell, Tiffany A / Velusamy, Thilaga / Tseng, Yeu-Yang / Tscharke, David C

The Journal of general virology

2018 Volume 99, Issue 5, Page(s) 682–692

Abstract: ... ...

Abstract	CD8
MeSH term(s)	Animals ; Antigen Presentation ; CD8-Positive T-Lymphocytes/immunology ; CD8-Positive T-Lymphocytes/virology ; Disease Models, Animal ; Female ; Ganglia, Sensory/virology ; Herpes Simplex/immunology ; Herpesvirus 1, Human/genetics ; Herpesvirus 1, Human/immunology ; Histocompatibility Antigens Class I ; Immunodominant Epitopes/genetics ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Skin/immunology ; Skin/pathology ; Skin/virology ; Specific Pathogen-Free Organisms ; Viral Load ; Virus Latency/immunology
Chemical Substances	Histocompatibility Antigens Class I ; Immunodominant Epitopes
Language	English
Publishing date	2018-04-05
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	219316-4
ISSN	1465-2099 ; 0022-1317
ISSN (online)	1465-2099
ISSN	0022-1317
DOI	10.1099/jgv.0.001059
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