Induction of DNA double-strand breaks and cellular senescence by human respiratory syncytial virus

ABSTRACT

Human respiratory syncytial virus (HRSV) accounts for the majority of lower respiratory tract infections during infancy and childhood and is associated with significant morbidity and mortality. HRSV provokes a proliferation arrest and characteristic syncytia in cellular systems such as immortalized epithelial cells. We show here that HRSV induces the expression of DNA damage markers and proliferation arrest such as P-TP53, P-ATM, CDKN1A and γH2AFX in cultured cells secondary to the production of mitochondrial reactive oxygen species (ROS). The DNA damage foci contained γH2AFX and TP53BP1, indicative of double-strand breaks (DSBs) and could be reversed by antioxidant treatments such as N-Acetylcysteine (NAC) or reduced glutathione ethyl ester (GSHee). The damage observed is associated with the accumulation of senescent cells, displaying a canonical senescent phenotype in both mononuclear cells and syncytia. In addition, we show signs of DNA damage and aging such as γH2AFX and CDKN2A expression in the respiratory epithelia of infected mice long after viral clearance. Altogether, these results show that HRSV triggers a DNA damage-mediated cellular senescence program probably mediated by oxidative stress. The results also suggest that this program might contribute to the physiopathology of the infection, tissue remodeling and aging, and might be associated to long-term consequences of HRSV infections.

KEYWORDS:

• cellular senescence
• DNA damage
• human respiratory
• ROS
• syncytial virus

This article is referred to by:

Increasing the complexity of respiratory syncytial virus infection: Reactive oxygen species, DNA damage, and premature senescence

Disclosure of potential conflicts of interest

No potential conflicts of interest were disclosed.

Acknowledgments

We thank the core facilities of the Instituto de Investigaciones Biomédicas de Madrid and of the Centro Nacional de Microbiología for technical help.

Author contributions

IM performed and designed experiments. V G-C performed and designed experiments. TG, A G-G and DG performed experiments, AA designed experiments and wrote paper. AZ performed experiments, designed experiments, wrote paper and conceived the project.

Funding

This work was supported by Grants MPY-1038/14 to Alberto Zambrano, PI 11/00590 to Isidoro Martínez and RD12/0036/0030 to Ana Aranda from FIS (Instituto de Salud Carlos III) and Grant BFU2011-28958, from Ministerio de Economía y Competitividad to Ana Aranda. The authors have no conflicting financial interests.