LRP6 Is a Functional Receptor for Attenuated Canine Distemper Virus
ABSTRACT Wild-type canine distemper virus (CDV) is an important pathogen of dogs as well as wildlife that can infect immune and epithelial cells through two known receptors: the signaling lymphocytic activation molecule (SLAM) and nectin-4, respectively. Conversely, the ferret and egg-adapted CDV-On...
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| Format: | Article |
| Language: | English |
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American Society for Microbiology
2023-02-01
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| Series: | mBio |
| Subjects: | |
| Online Access: | https://journals.asm.org/doi/10.1128/mbio.03114-22 |
| _version_ | 1811163381482127360 |
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| author | Vaiva Gradauskaite Marine Inglebert John Doench Melanie Scherer Martina Dettwiler Marianne Wyss Neeta Shrestha Sven Rottenberg Philippe Plattet |
| author_facet | Vaiva Gradauskaite Marine Inglebert John Doench Melanie Scherer Martina Dettwiler Marianne Wyss Neeta Shrestha Sven Rottenberg Philippe Plattet |
| author_sort | Vaiva Gradauskaite |
| collection | DOAJ |
| description | ABSTRACT Wild-type canine distemper virus (CDV) is an important pathogen of dogs as well as wildlife that can infect immune and epithelial cells through two known receptors: the signaling lymphocytic activation molecule (SLAM) and nectin-4, respectively. Conversely, the ferret and egg-adapted CDV-Onderstepoort strain (CDV-OP) is employed as an effective vaccine for dogs. CDV-OP also exhibits promising oncolytic properties, such as its abilities to infect and kill multiple cancer cells in vitro. Interestingly, several cancer cells do not express SLAM or nectin-4, suggesting the presence of a yet unknown entry factor for CDV-OP. By conducting a genome-wide CRISPR/Cas9 knockout (KO) screen in CDV-OP-susceptible canine mammary carcinoma P114 cells, which neither express SLAM nor nectin-4, we identified low-density lipoprotein receptor-related protein 6 (LRP6) as a host factor that promotes CDV-OP infectivity. Whereas the genetic ablation of LRP6 rendered cells resistant to infection, ectopic expression in resistant LRP6KO cells restored susceptibility. Furthermore, multiple functional studies revealed that (i) the overexpression of LRP6 leads to increased cell-cell fusion, (ii) a soluble construct of the viral receptor-binding protein (solHOP) interacts with a soluble form of LRP6 (solLRP6), (iii) an H-OP point mutant that prevents interaction with solLRP6 abrogates cell entry in multiple cell lines once transferred into recombinant viral particles, and (iv) vesicular stomatitis virus (VSV) pseudotyped with CDV-OP envelope glycoproteins loses its infectivity in LRP6KO cells. Collectively, our study identified LRP6 as the long sought-after cell entry receptor of CDV-OP in multiple cell lines, which set the molecular bases to refine our understanding of viral-cell adaptation and to further investigate its oncolytic properties. IMPORTANCE Oncolytic viruses (OV) have gathered increasing interest in recent years as an alternative option to treat cancers. The Onderstepoort strain of canine distemper virus (CDV-OP), an enveloped RNA virus belonging to the genus Morbillivirus, is employed as a safe and efficient vaccine for dogs against distemper disease. Importantly, although CDV-OP can infect and kill multiple cancer cell lines, the basic mechanisms of entry remain to be elucidated, as most of those transformed cells do not express natural receptors (i.e., SLAM and nectin-4). In this study, using a genome-wide CRISPR/Cas9 knockout screen, we describe the discovery of LRP6 as a novel functional entry receptor for CDV-OP in various cancer cell lines and thereby uncover a basic mechanism of cell culture adaptation. Since LRP6 is upregulated in various cancer types, our data provide important insights in order to further investigate the oncolytic properties of CDV-OP. |
| first_indexed | 2024-04-10T06:44:09Z |
| format | Article |
| id | doaj.art-8b943239a2044dd48e7f51910c03fde1 |
| institution | Directory Open Access Journal |
| issn | 2150-7511 |
| language | English |
| last_indexed | 2024-04-10T06:44:09Z |
| publishDate | 2023-02-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | mBio |
| spelling | doaj.art-8b943239a2044dd48e7f51910c03fde12023-02-28T14:06:24ZengAmerican Society for MicrobiologymBio2150-75112023-02-0114110.1128/mbio.03114-22LRP6 Is a Functional Receptor for Attenuated Canine Distemper VirusVaiva Gradauskaite0Marine Inglebert1John Doench2Melanie Scherer3Martina Dettwiler4Marianne Wyss5Neeta Shrestha6Sven Rottenberg7Philippe Plattet8Division of Neurological Sciences, Vetsuisse Faculty, University of Bern, Bern, SwitzerlandInstitute of Animal Pathology, Vetsuisse Faculty, University of Bern, Bern, SwitzerlandGenetic Perturbation Platform, Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USADivision of Neurological Sciences, Vetsuisse Faculty, University of Bern, Bern, SwitzerlandInstitute of Animal Pathology, Vetsuisse Faculty, University of Bern, Bern, SwitzerlandDivision of Neurological Sciences, Vetsuisse Faculty, University of Bern, Bern, SwitzerlandDivision of Neurological Sciences, Vetsuisse Faculty, University of Bern, Bern, SwitzerlandInstitute of Animal Pathology, Vetsuisse Faculty, University of Bern, Bern, SwitzerlandDivision of Neurological Sciences, Vetsuisse Faculty, University of Bern, Bern, SwitzerlandABSTRACT Wild-type canine distemper virus (CDV) is an important pathogen of dogs as well as wildlife that can infect immune and epithelial cells through two known receptors: the signaling lymphocytic activation molecule (SLAM) and nectin-4, respectively. Conversely, the ferret and egg-adapted CDV-Onderstepoort strain (CDV-OP) is employed as an effective vaccine for dogs. CDV-OP also exhibits promising oncolytic properties, such as its abilities to infect and kill multiple cancer cells in vitro. Interestingly, several cancer cells do not express SLAM or nectin-4, suggesting the presence of a yet unknown entry factor for CDV-OP. By conducting a genome-wide CRISPR/Cas9 knockout (KO) screen in CDV-OP-susceptible canine mammary carcinoma P114 cells, which neither express SLAM nor nectin-4, we identified low-density lipoprotein receptor-related protein 6 (LRP6) as a host factor that promotes CDV-OP infectivity. Whereas the genetic ablation of LRP6 rendered cells resistant to infection, ectopic expression in resistant LRP6KO cells restored susceptibility. Furthermore, multiple functional studies revealed that (i) the overexpression of LRP6 leads to increased cell-cell fusion, (ii) a soluble construct of the viral receptor-binding protein (solHOP) interacts with a soluble form of LRP6 (solLRP6), (iii) an H-OP point mutant that prevents interaction with solLRP6 abrogates cell entry in multiple cell lines once transferred into recombinant viral particles, and (iv) vesicular stomatitis virus (VSV) pseudotyped with CDV-OP envelope glycoproteins loses its infectivity in LRP6KO cells. Collectively, our study identified LRP6 as the long sought-after cell entry receptor of CDV-OP in multiple cell lines, which set the molecular bases to refine our understanding of viral-cell adaptation and to further investigate its oncolytic properties. IMPORTANCE Oncolytic viruses (OV) have gathered increasing interest in recent years as an alternative option to treat cancers. The Onderstepoort strain of canine distemper virus (CDV-OP), an enveloped RNA virus belonging to the genus Morbillivirus, is employed as a safe and efficient vaccine for dogs against distemper disease. Importantly, although CDV-OP can infect and kill multiple cancer cell lines, the basic mechanisms of entry remain to be elucidated, as most of those transformed cells do not express natural receptors (i.e., SLAM and nectin-4). In this study, using a genome-wide CRISPR/Cas9 knockout screen, we describe the discovery of LRP6 as a novel functional entry receptor for CDV-OP in various cancer cell lines and thereby uncover a basic mechanism of cell culture adaptation. Since LRP6 is upregulated in various cancer types, our data provide important insights in order to further investigate the oncolytic properties of CDV-OP.https://journals.asm.org/doi/10.1128/mbio.03114-22attenuated CDVCRISPR/Cas9 KO screenLRP6cell entryreceptor |
| spellingShingle | Vaiva Gradauskaite Marine Inglebert John Doench Melanie Scherer Martina Dettwiler Marianne Wyss Neeta Shrestha Sven Rottenberg Philippe Plattet LRP6 Is a Functional Receptor for Attenuated Canine Distemper Virus mBio attenuated CDV CRISPR/Cas9 KO screen LRP6 cell entry receptor |
| title | LRP6 Is a Functional Receptor for Attenuated Canine Distemper Virus |
| title_full | LRP6 Is a Functional Receptor for Attenuated Canine Distemper Virus |
| title_fullStr | LRP6 Is a Functional Receptor for Attenuated Canine Distemper Virus |
| title_full_unstemmed | LRP6 Is a Functional Receptor for Attenuated Canine Distemper Virus |
| title_short | LRP6 Is a Functional Receptor for Attenuated Canine Distemper Virus |
| title_sort | lrp6 is a functional receptor for attenuated canine distemper virus |
| topic | attenuated CDV CRISPR/Cas9 KO screen LRP6 cell entry receptor |
| url | https://journals.asm.org/doi/10.1128/mbio.03114-22 |
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