PMID- 32941510
OWN - NLM
STAT- MEDLINE
DCOM- 20201103
LR  - 20201103
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 15
IP  - 9
DP  - 2020
TI  - Reticulon-3 modulates the incorporation of replication competent hepatitis C 
      virus molecules for release inside infectious exosomes.
PG  - e0239153
LID - 10.1371/journal.pone.0239153 [doi]
LID - e0239153
AB  - 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 naive 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.
FAU - Li, Jingjing
AU  - Li J
AD  - INRS-Institut Armand-Frappier, Institut National de la Recherche Scientifique, 
      Laval, Quebec, Canada.
FAU - Abosmaha, Ebtisam
AU  - Abosmaha E
AD  - INRS-Institut Armand-Frappier, Institut National de la Recherche Scientifique, 
      Laval, Quebec, Canada.
FAU - Coffin, Carla S
AU  - Coffin CS
AUID- ORCID: 0000-0002-1472-0901
AD  - Department of Medicine, University of Calgary, Calgary, Alberta, Canada.
FAU - Labonte, Patrick
AU  - Labonte P
AD  - INRS-Institut Armand-Frappier, Institut National de la Recherche Scientifique, 
      Laval, Quebec, Canada.
FAU - Bukong, Terence Ndonyi
AU  - Bukong TN
AUID- ORCID: 0000-0003-3898-0617
AD  - INRS-Institut Armand-Frappier, Institut National de la Recherche Scientifique, 
      Laval, Quebec, Canada.
LA  - eng
GR  - CIHR/Canada
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20200917
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
RN  - 0 (Carrier Proteins)
RN  - 0 (Membrane Proteins)
RN  - 0 (Nerve Tissue Proteins)
RN  - 0 (RTN3 protein, human)
SB  - IM
MH  - Carrier Proteins/*metabolism
MH  - Cell Line
MH  - Exosomes/*metabolism/virology
MH  - Female
MH  - Hepacivirus/*physiology
MH  - Hepatitis C/*metabolism/virology
MH  - Host-Pathogen Interactions
MH  - Humans
MH  - Male
MH  - Membrane Proteins/*metabolism
MH  - Middle Aged
MH  - Nerve Tissue Proteins/*metabolism
PMC - PMC7498005
COIS- The authors declare that they have NO conflicts of interest
EDAT- 2020/09/18 06:00
MHDA- 2020/11/04 06:00
PMCR- 2020/09/17
CRDT- 2020/09/17 17:17
PHST- 2020/04/23 00:00 [received]
PHST- 2020/08/31 00:00 [accepted]
PHST- 2020/09/17 17:17 [entrez]
PHST- 2020/09/18 06:00 [pubmed]
PHST- 2020/11/04 06:00 [medline]
PHST- 2020/09/17 00:00 [pmc-release]
AID - PONE-D-20-11851 [pii]
AID - 10.1371/journal.pone.0239153 [doi]
PST - epublish
SO  - PLoS One. 2020 Sep 17;15(9):e0239153. doi: 10.1371/journal.pone.0239153. 
      eCollection 2020.