caa a22 4500 606177701 CHVBK 20210128100753.0 cr unu---uuuuu 210128e20150801xx s 000 0 eng 10.1007/s00018-015-1879-1 doi (NATIONALLICENCE)springer-10.1007/s00018-015-1879-1 Influenza infection induces host DNA damage and dynamic DNA damage responses during tissue regeneration [Elektronische Daten] [Na Li, Marcus Parrish, Tze Chan, Lu Yin, Prashant Rai, Yamada Yoshiyuki, Nona Abolhassani, Kong Tan, Orsolya Kiraly, Vincent Chow, Bevin Engelward] Influenza viruses account for significant morbidity worldwide. Inflammatory responses, including excessive generation of reactive oxygen and nitrogen species (RONS), mediate lung injury in severe influenza infections. However, the molecular basis of inflammation-induced lung damage is not fully understood. Here, we studied influenza H1N1 infected cells in vitro, as well as H1N1 infected mice, and we monitored molecular and cellular responses over the course of 2weeks in vivo. We show that influenza induces DNA damage to both, when cells are directly exposed to virus in vitro (measured using the comet assay) and also when cells are exposed to virus in vivo (estimated via γH2AX foci). We show that DNA damage, as well as responses to DNA damage persist in vivo until long after virus has been cleared, at times when there are inflammation associated RONS (measured by xanthine oxidase activity and oxidative products). The frequency of lung epithelial and immune cells with increased γH2AX foci is elevated in vivo, especially for dividing cells (Ki-67-positive) exposed to oxidative stress during tissue regeneration. Additionally, we observed a significant increase in apoptotic cells as well as increased levels of DNA double strand break (DSB) repair proteins Ku70, Ku86 and Rad51 during the regenerative phase. In conclusion, results show that influenza induces DNA damageboth in vitro and in vivo, and that DNA damage responses are activated, raising the possibility that DNA repair capacity may be a determining factor for tissue recovery and disease outcome. Springer Basel, 2015 Nuclear foci nationallicence Immunofluorescence nationallicence Repair deficiency nationallicence Acute infection nationallicence εdA : 1, N6-Etheno-2′-deoxyadenosine nationallicence εdG : 1, N2-Etheno-2′-deoxyguanosine nationallicence 8-OH-dG : 8-Hydroxy-deoxyguanosine nationallicence 8-OH-G : 8-Hydroxyguanosine nationallicence AEII : Alveolar epithelial type II cells nationallicence ATM : Ataxia telangiectasia mutated nationallicence ATR : ATM- and Rad-3 related nationallicence BER : Base excision repair nationallicence BALF : Bronchoalveolar lavage fluid nationallicence CCSP : Club cell secretary protein nationallicence DDR : DNA damage response nationallicence DSBs : DNA double-strand breaks nationallicence DNA-PKcs : DNA-dependent protein kinase catalytic subunit nationallicence HA : Hemagglutinin nationallicence HR : Homologous recombination nationallicence MDCK : Madin-Darby canine kidney nationallicence MOI : Multiplicity of infection nationallicence NHEJ : Non-homologous end joining nationallicence NS1 : Non-structural protein 1 nationallicence PI3K-like kinases : Phosphatidylinositol-3-kinase-like kinases nationallicence Pro-SPC : Pro-surfactant protein C nationallicence RONS : Reactive oxygen and nitrogen species nationallicence SSBs : DNA single strand breaks nationallicence XO : Xanthine oxidase nationallicence Li Na Singapore-MIT Alliance for Research and Technology, 1 CREATE Way, #03-10/11 Innovation Wing, #03-12/13/14 Enterprise Wing, 138602, Singapore, Singapore aut Parrish Marcus Department of Biological Engineering, Massachusetts Institute of Technology, 77 Massachusetts Ave., 16-743, 02139, Cambridge, MA, USA aut Chan Tze Singapore-MIT Alliance for Research and Technology, 1 CREATE Way, #03-10/11 Innovation Wing, #03-12/13/14 Enterprise Wing, 138602, Singapore, Singapore aut Yin Lu Singapore-MIT Alliance for Research and Technology, 1 CREATE Way, #03-10/11 Innovation Wing, #03-12/13/14 Enterprise Wing, 138602, Singapore, Singapore aut Rai Prashant Singapore-MIT Alliance for Research and Technology, 1 CREATE Way, #03-10/11 Innovation Wing, #03-12/13/14 Enterprise Wing, 138602, Singapore, Singapore aut Yoshiyuki Yamada Singapore-MIT Alliance for Research and Technology, 1 CREATE Way, #03-10/11 Innovation Wing, #03-12/13/14 Enterprise Wing, 138602, Singapore, Singapore aut Abolhassani Nona Department of Biological Engineering, Massachusetts Institute of Technology, 77 Massachusetts Ave., 16-743, 02139, Cambridge, MA, USA aut Tan Kong Department of Pathology, Yong loo Lin School of Medicine, National University Health System and National University of Singapore, Lower Kent Ridge Road, 119074, Singapore, Singapore aut Kiraly Orsolya Singapore-MIT Alliance for Research and Technology, 1 CREATE Way, #03-10/11 Innovation Wing, #03-12/13/14 Enterprise Wing, 138602, Singapore, Singapore aut Chow Vincent Department of Microbiology, National University of Singapore, 5 Science Drive 2, Blk MD4, Level 3, 117545, Singapore, Singapore aut Engelward Bevin Department of Biological Engineering, Massachusetts Institute of Technology, 77 Massachusetts Ave., 16-743, 02139, Cambridge, MA, USA aut Cellular and Molecular Life Sciences Springer Basel 72/15(2015-08-01), 2973-2988 1420-682X 72:15<2973 2015 72 18 https://doi.org/10.1007/s00018-015-1879-1 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00018-015-1879-1 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Li Na Singapore-MIT Alliance for Research and Technology, 1 CREATE Way, #03-10/11 Innovation Wing, #03-12/13/14 Enterprise Wing, 138602, Singapore, Singapore aut NATIONALLICENCE 700 1- Parrish Marcus Department of Biological Engineering, Massachusetts Institute of Technology, 77 Massachusetts Ave., 16-743, 02139, Cambridge, MA, USA aut NATIONALLICENCE 700 1- Chan Tze Singapore-MIT Alliance for Research and Technology, 1 CREATE Way, #03-10/11 Innovation Wing, #03-12/13/14 Enterprise Wing, 138602, Singapore, Singapore aut NATIONALLICENCE 700 1- Yin Lu Singapore-MIT Alliance for Research and Technology, 1 CREATE Way, #03-10/11 Innovation Wing, #03-12/13/14 Enterprise Wing, 138602, Singapore, Singapore aut NATIONALLICENCE 700 1- Rai Prashant Singapore-MIT Alliance for Research and Technology, 1 CREATE Way, #03-10/11 Innovation Wing, #03-12/13/14 Enterprise Wing, 138602, Singapore, Singapore aut NATIONALLICENCE 700 1- Yoshiyuki Yamada Singapore-MIT Alliance for Research and Technology, 1 CREATE Way, #03-10/11 Innovation Wing, #03-12/13/14 Enterprise Wing, 138602, Singapore, Singapore aut NATIONALLICENCE 700 1- Abolhassani Nona Department of Biological Engineering, Massachusetts Institute of Technology, 77 Massachusetts Ave., 16-743, 02139, Cambridge, MA, USA aut NATIONALLICENCE 700 1- Tan Kong Department of Pathology, Yong loo Lin School of Medicine, National University Health System and National University of Singapore, Lower Kent Ridge Road, 119074, Singapore, Singapore aut NATIONALLICENCE 700 1- Kiraly Orsolya Singapore-MIT Alliance for Research and Technology, 1 CREATE Way, #03-10/11 Innovation Wing, #03-12/13/14 Enterprise Wing, 138602, Singapore, Singapore aut NATIONALLICENCE 700 1- Chow Vincent Department of Microbiology, National University of Singapore, 5 Science Drive 2, Blk MD4, Level 3, 117545, Singapore, Singapore aut NATIONALLICENCE 700 1- Engelward Bevin Department of Biological Engineering, Massachusetts Institute of Technology, 77 Massachusetts Ave., 16-743, 02139, Cambridge, MA, USA aut NATIONALLICENCE 773 0- Cellular and Molecular Life Sciences Springer Basel 72/15(2015-08-01), 2973-2988 1420-682X 72:15<2973 2015 72 18