Reticulon-3 modulates the incorporation of replication competent hepatitis C virus molecules for release inside infectious exosomes
Background Cell released microvesicles specifically, exosomes, play an important role in mediating immunologic escape, treatment resistance, and disease persistence of Hepatitis C virus (HCV) infection. Reports on the molecular compositions of exosomes released by cells under diverse conditions, esp...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2020-01-01
|
| Series: | PLoS ONE |
| Online Access: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7498005/?tool=EBI |
| _version_ | 1819028418711257088 |
|---|---|
| author | Jingjing Li Ebtisam Abosmaha Carla S. Coffin Patrick Labonté Terence Ndonyi Bukong Rafael Aldabe |
| author_facet | Jingjing Li Ebtisam Abosmaha Carla S. Coffin Patrick Labonté Terence Ndonyi Bukong Rafael Aldabe |
| author_sort | Jingjing Li |
| collection | DOAJ |
| description | Background Cell released microvesicles specifically, exosomes, play an important role in mediating immunologic escape, treatment resistance, and disease persistence of Hepatitis C virus (HCV) infection. Reports on the molecular compositions of exosomes released by cells under diverse conditions, especially during viral infections, suggest that their cargo contents are not randomly loaded. However, the precise molecular mechanisms directing the selective cargo sorting and loading inside infectious viral exosomes remains elusive. Aim To decipher the role of Reticulon 3 (RTN3) in the selective molecular cargo sorting and loading inside infectious viral exosomes during HCV infection. Methods We used Huh7 cells–JFH1 HCV infection and HCV Full-Length (FL) replicon systems. Additionally, we analyzed human liver and serum exosome samples from healthy and treatment naïve HCV infected individuals. Our experiments made use of molecular biology and immunology techniques. Results HCV infection (JFH1-Huh7 or HCV-FL replicon cells) was associated with increased RTN3L&S isoforms expression in cells and cell released exosomes. Accordingly, increased expression of RTN3L&S was observed in liver and serum exosome samples of HCV infected individuals compared to healthy controls. RNA-ChIP analysis revealed that RTN3L&S interacted with dsHCV RNA. Lentiviral CRISPR/Cas9 mediated knockdown (KD) of RTN3 and plasmid overexpression (OE) of wild type, C- and N-terminal deletion mutants of RTN3L&S in HCV- infected Huh7 cells differentially impacted the cellular release of infectious viral exosomes. RTN3L&S KD significantly decreased, while RTN3S OE significantly increased the number of Huh7 cell-released infectious exosomes. The C-terminal domain of RTN3 interacted with and modulated the loading of dsHCV RNA inside infectious exosomes. Antiviral treatment of HCV infected Huh7 cells reduced virus-induced RTN3L&S expression and attenuated the release of infectious exosomes. Conclusion RTN3 constitutes a novel regulator and a potential therapeutic target that mediates the specific loading of infectious viral exosomes. |
| first_indexed | 2024-12-21T05:58:03Z |
| format | Article |
| id | doaj.art-41f3cf10c3044f2cadcff5c2ed25f8f2 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-21T05:58:03Z |
| publishDate | 2020-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-41f3cf10c3044f2cadcff5c2ed25f8f22022-12-21T19:13:49ZengPublic Library of Science (PLoS)PLoS ONE1932-62032020-01-01159Reticulon-3 modulates the incorporation of replication competent hepatitis C virus molecules for release inside infectious exosomesJingjing LiEbtisam AbosmahaCarla S. CoffinPatrick LabontéTerence Ndonyi BukongRafael AldabeBackground Cell released microvesicles specifically, exosomes, play an important role in mediating immunologic escape, treatment resistance, and disease persistence of Hepatitis C virus (HCV) infection. Reports on the molecular compositions of exosomes released by cells under diverse conditions, especially during viral infections, suggest that their cargo contents are not randomly loaded. However, the precise molecular mechanisms directing the selective cargo sorting and loading inside infectious viral exosomes remains elusive. Aim To decipher the role of Reticulon 3 (RTN3) in the selective molecular cargo sorting and loading inside infectious viral exosomes during HCV infection. Methods We used Huh7 cells–JFH1 HCV infection and HCV Full-Length (FL) replicon systems. Additionally, we analyzed human liver and serum exosome samples from healthy and treatment naïve HCV infected individuals. Our experiments made use of molecular biology and immunology techniques. Results HCV infection (JFH1-Huh7 or HCV-FL replicon cells) was associated with increased RTN3L&S isoforms expression in cells and cell released exosomes. Accordingly, increased expression of RTN3L&S was observed in liver and serum exosome samples of HCV infected individuals compared to healthy controls. RNA-ChIP analysis revealed that RTN3L&S interacted with dsHCV RNA. Lentiviral CRISPR/Cas9 mediated knockdown (KD) of RTN3 and plasmid overexpression (OE) of wild type, C- and N-terminal deletion mutants of RTN3L&S in HCV- infected Huh7 cells differentially impacted the cellular release of infectious viral exosomes. RTN3L&S KD significantly decreased, while RTN3S OE significantly increased the number of Huh7 cell-released infectious exosomes. The C-terminal domain of RTN3 interacted with and modulated the loading of dsHCV RNA inside infectious exosomes. Antiviral treatment of HCV infected Huh7 cells reduced virus-induced RTN3L&S expression and attenuated the release of infectious exosomes. Conclusion RTN3 constitutes a novel regulator and a potential therapeutic target that mediates the specific loading of infectious viral exosomes.https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7498005/?tool=EBI |
| spellingShingle | Jingjing Li Ebtisam Abosmaha Carla S. Coffin Patrick Labonté Terence Ndonyi Bukong Rafael Aldabe Reticulon-3 modulates the incorporation of replication competent hepatitis C virus molecules for release inside infectious exosomes PLoS ONE |
| title | Reticulon-3 modulates the incorporation of replication competent hepatitis C virus molecules for release inside infectious exosomes |
| title_full | Reticulon-3 modulates the incorporation of replication competent hepatitis C virus molecules for release inside infectious exosomes |
| title_fullStr | Reticulon-3 modulates the incorporation of replication competent hepatitis C virus molecules for release inside infectious exosomes |
| title_full_unstemmed | Reticulon-3 modulates the incorporation of replication competent hepatitis C virus molecules for release inside infectious exosomes |
| title_short | Reticulon-3 modulates the incorporation of replication competent hepatitis C virus molecules for release inside infectious exosomes |
| title_sort | reticulon 3 modulates the incorporation of replication competent hepatitis c virus molecules for release inside infectious exosomes |
| url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7498005/?tool=EBI |
| work_keys_str_mv | AT jingjingli reticulon3modulatestheincorporationofreplicationcompetenthepatitiscvirusmoleculesforreleaseinsideinfectiousexosomes AT ebtisamabosmaha reticulon3modulatestheincorporationofreplicationcompetenthepatitiscvirusmoleculesforreleaseinsideinfectiousexosomes AT carlascoffin reticulon3modulatestheincorporationofreplicationcompetenthepatitiscvirusmoleculesforreleaseinsideinfectiousexosomes AT patricklabonte reticulon3modulatestheincorporationofreplicationcompetenthepatitiscvirusmoleculesforreleaseinsideinfectiousexosomes AT terencendonyibukong reticulon3modulatestheincorporationofreplicationcompetenthepatitiscvirusmoleculesforreleaseinsideinfectiousexosomes AT rafaelaldabe reticulon3modulatestheincorporationofreplicationcompetenthepatitiscvirusmoleculesforreleaseinsideinfectiousexosomes |