Structural
Changes In Cells Imaged by Soft X‑ray
Cryo-Tomography During Hepatitis C Virus Infection
Ana Joaquina Pérez-Berná
Maria
José Rodríguez
Francisco Javier Chichón
Martina
Friederike Friesland
Andrea Sorrentino
Jose L. Carrascosa
Eva Pereiro
Pablo Gastaminza
10.1021/acsnano.6b01374.s002
https://acs.figshare.com/articles/media/Structural_Changes_In_Cells_Imaged_by_Soft_X_ray_Cryo-Tomography_During_Hepatitis_C_Virus_Infection/3465944
Chronic
hepatitis C virus (HCV) infection causes severe liver disease
in millions of humans worldwide. Pathogenesis of HCV infection is
strongly driven by a deficient immune response of the host, although
intersection of different aspects of the virus life cycle with cellular
homeostasis is emerging as an important player in the pathogenesis
and progression of the disease. Cryo soft X-ray tomography (cryo-SXT)
was performed to investigate the ultrastructural alterations induced
by the interference of HCV replication with cellular homeostasis.
Native, whole cell, three-dimensional (3D) maps were obtained in HCV
replicon-harboring cells and in a surrogate model of HCV infection.
Tomograms from HCV-replicating cells show blind-ended endoplasmic
reticulum tubules with pseudospherical extrusions and marked alterations
of mitochondrial morphology that correlated spatially with the presence
of endoplasmic reticulum alterations, suggesting a short-range influence
of the viral machinery on mitochondrial homeostasis. Both mitochondrial
and endoplasmic reticulum alterations could be reverted by a combination
of sofosbuvir/daclatasvir, which are clinically approved direct-acting
antivirals for the treatment of chronic HCV infection. In addition
to providing structural insight into cellular aspects of HCV pathogenesis,
our study illustrates how cryo-SXT is a powerful 3D wide-field imaging
tool for the assessment and understanding of complex cellular processes
in a setting of near-native whole hydrated cells. Our results also
constitute a proof of concept for the use of cryo-SXT as a platform
that enables determining the potential impact of candidate compounds
on the ultrastructure of the cell that may assist drug development
at a preclinical level.
2016-06-21 00:00:00
Hepatitis C Virus Infection Chronic hepatitis C virus
homeostasi
endoplasmic reticulum alterations
virus life cycle
mitochondrial
HCV infection