Direct imaging of the disruption of hepatitis C virus replication complexes by inhibitors of lipid metabolism.
Here we have simultaneously characterized the influence of inhibitors of peroxisome proliferator-activated receptor alpha (PPARalpha) and the mevalonate pathway on hepatocyte lipid metabolism and the subcellular localization of hepatitis C virus (HCV) RNA using two-photon fluorescence (TPF) and coherent anti-Stokes Raman scattering (CARS) microscopy. Using this approach, we demonstrate that modulators of PPARalpha signaling rapidly cause the dispersion of HCV RNA from replication sites and simultaneously induce lipid storage and increases in lipid droplet size. We demonstrate that reductions in the levels of cholesterol resulting from inhibition of the mevalonate pathway upregulates triglyceride levels. We also show that the rate of dispersion of HCV RNA is very rapid when using a PPARalpha antagonist. This occurs with a faster rate to that of direct inhibition of 3-hydroxy-3-methyglutaryl CoA reductase (HMG-CoA reductase) using lovastatin in living cells, demonstrating the potential therapeutic value of modulating host cell pathways as part of a strategy to eliminate chronic HCV infection.
Lyn
Rodney K
RK
Steacie Institute for Molecular Sciences, National Research Council of Canada, 100 Sussex Drive, Ottawa, Canada K1A 0R6.
Kennedy
David C
DC
Sagan
Selena M
SM
Blais
David R
DR
Rouleau
Yanouchka
Y
Pegoraro
Adrian F
AF
Xie
X Sunney
XS
Stolow
Albert
A
Pezacki
John Paul
JP
D002460:Cell Line; D016174:Hepacivirus / Q000187:drug effects / Q000502:physiology*; D022781:Hepatocytes / Q000187:drug effects* / Q000378:metabolism / Q000821:virology; D006801:Humans; D050356:Lipid Metabolism / Q000187:drug effects*; D008856:Microscopy, Fluorescence; D047493:PPAR alpha / Q000037:antagonists & inhibitors*; D012367:RNA, Viral / Q000378:metabolism; D013059:Spectrum Analysis, Raman; D014779:Virus Replication / Q000187:drug effects*
D047493:PPAR alpha; D012367:RNA, Viral