AI Article Synopsis
- Exosomes released by cells during Hepatitis C virus (HCV) infection play a crucial role in immune evasion and treatment resistance, but the specific mechanisms behind how they sort and load their molecular cargo remains unclear.
- The study aims to investigate how Reticulon 3 (RTN3) influences the selective sorting and loading of molecules into these exosomes during HCV infection.
- Results show that RTN3 expression increases in HCV-infected cells and exosomes, and manipulating RTN3 levels affects the quantity of infectious viral exosomes released, suggesting its key role in the process.
Article Abstract
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.
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Source |
|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7498005 | PMC |
| http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0239153 | PLOS |
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