The first versatile human iPSC-based model of ectopic virus induction allows new insights in RNA-virus disease

Abstract

A detailed description of pathophysiological effects that viruses exert on their host is still challenging. For the first time, we report a highly controllable viral expression model based on an iPS-cell line from a healthy human donor. The established viral model system enables a dose-dependent and highly localized RNA-virus expression in a fully controllable environment, giving rise for new applications for the scientific community.

The First Viral Expression Model in Human Induced Pluripotent Stem Cells to Observe Enteroviral Infections In vitro

“…Transport of these viral replication complexes to the cell membrane and additional membrane destabilization finally leads to the release of viral progeny to infect neighbouring cells. Obvious membrane destabilization of the cellular membrane and of the membrane of cytoplasmatic structures was observed by Peischard et al in hiPSC-derived cardiomyocytes as well [3]. While no viral progeny is produced during CVB3∆VP0 expression, the effects of the expressed viral proteins potentially lead to the formation of viral replication complexes and to the destabilization of the cell membrane.…”

“…An infection of hostcells with CVB3 wildtypes was found to be associated with the elevation of mitochondrial reactive oxygen species (ROS) together with pathologic mitochondrial phenotypes [12]. The elevation of mitochondrial ROS-levels in hiPSCderived cardiomyocytes expressing CVB3∆VP0 verified the functionality of the system [3]. The direct effects of these ROS are still unclear and hard to study.…”

“…This binding enables gene expression as in this case of CVB3∆VP0. Thereby, the experimenter can adjust the desired virus load by titrating the doxycycline concentration applied to the cells [3]. This dose-dependence of this system enables for the simulation of various infection scenario from strong, acute infections to chronic infections with long-lasting, low viral loads.…”

“…Due to rapid apoptosis of cells infected with CVB3 wildtype strains makes a detailed view on mitochondrial changes almost impossible and long-term, chronical infections are non-observable. In the hiPSC-based expression system of Peischard et al no significant elevation of apoptosis was detected after CVB3∆VP0 expression, because no competent viruses are produced [3]. This enables for long-term observation of mitochondrial effects induced by CVB3∆VP0.…”

“…Although, CVB3 is long-time known to cause these pathologic phenotypes, less is known about the molecular interactions of CVB3 and its host-cell. Recently, a new concept for viral model systems was developed, which allows the detailed observation of viruses, here CVB3, in human iPSC-derived cells [3]. The genome of a non-infectious variant of CVB3, CVB3∆VP0, was inserted into the hiPSC-genome via PiggyBac transposase together with a doxycyclineinducible tet-on system.…”