Yi-Han Lin1, Maryann P Platt1, Ryan P Gilley2, David Brown1, Peter H Dube2, Yanbao Yu1, Norberto Gonzalez-Juarbe1. 1. Infectious Diseases and Genomic Medicine Group, J Craig Venter Institute, Rockville, MD (Y.-H.L., M.P.P., D.B., Y.Y., N.G.-J.). 2. Department of Microbiology, Immunology and Molecular Genetics, The University of Texas Health Science Center at San Antonio, TX (R.P.G., P.H.D.).
Abstract
RATIONALE: Patients with and without cardiovascular diseases have been shown to be at risk of influenza-mediated cardiac complications. Recent clinical reports support the notion of a direct link between laboratory-confirmed influenza virus infections and adverse cardiac events. OBJECTIVE: Define the molecular mechanisms underlying influenza virus-induced cardiac pathogenesis after resolution of pulmonary infection and the role of necroptosis in this process. METHODS AND RESULTS: Hearts from wild-type and necroptosis-deficient (MLKL [mixed lineage kinase domain-like protein]-KO) mice were dissected 12 days after initial influenza A virus (IAV) infection when viral titers were undetectable in the lungs. Immunofluorescence microscopy and plaque assays showed presence of viable IAV particles in the myocardium without generation of interferon responses. Global proteome and phosphoproteome analyses using high-resolution accurate mass-based LC-MS/MS and label-free quantitation showed that the global proteome as well as the phosphoproteome profiles were significantly altered in IAV-infected mouse hearts in a strain-independent manner. Necroptosis-deficient mice had increased survival and reduced weight loss post-IAV infection, as well as increased antioxidant and mitochondrial function, indicating partial protection to IAV infection. These findings were confirmed in vitro by pretreatment of human and rat myocytes with antioxidants or necroptosis inhibitors, which blunted oxidative stress and mitochondrial damage after IAV infection. CONCLUSIONS: This study provides the first evidence that the cardiac proteome and phosphoproteome are significantly altered post-pulmonary influenza infection. Moreover, viral particles can persist in the heart after lung clearance, altering mitochondrial function and promoting cell death without active replication and interferon responses. Finally, our findings show inhibition of necroptosis or prevention of mitochondrial damage as possible therapeutic interventions to reduce cardiac damage during influenza infections. Graphic Abstract: A graphic abstract is available for this article.
RATIONALE: Patients with and without cardiovascular diseases have been shown to be at risk of influenza-mediated cardiac complications. Recent clinical reports support the notion of a direct link between laboratory-confirmed influenza virus infections and adverse cardiac events. OBJECTIVE: Define the molecular mechanisms underlying influenza virus-induced cardiac pathogenesis after resolution of pulmonary infection and the role of necroptosis in this process. METHODS AND RESULTS: Hearts from wild-type and necroptosis-deficient (MLKL [mixed lineage kinase domain-like protein]-KO) mice were dissected 12 days after initial influenza A virus (IAV) infection when viral titers were undetectable in the lungs. Immunofluorescence microscopy and plaque assays showed presence of viable IAV particles in the myocardium without generation of interferon responses. Global proteome and phosphoproteome analyses using high-resolution accurate mass-based LC-MS/MS and label-free quantitation showed that the global proteome as well as the phosphoproteome profiles were significantly altered in IAV-infected mouse hearts in a strain-independent manner. Necroptosis-deficient mice had increased survival and reduced weight loss post-IAV infection, as well as increased antioxidant and mitochondrial function, indicating partial protection to IAV infection. These findings were confirmed in vitro by pretreatment of human and rat myocytes with antioxidants or necroptosis inhibitors, which blunted oxidative stress and mitochondrial damage after IAV infection. CONCLUSIONS: This study provides the first evidence that the cardiac proteome and phosphoproteome are significantly altered post-pulmonary influenza infection. Moreover, viral particles can persist in the heart after lung clearance, altering mitochondrial function and promoting cell death without active replication and interferon responses. Finally, our findings show inhibition of necroptosis or prevention of mitochondrial damage as possible therapeutic interventions to reduce cardiac damage during influenza infections. Graphic Abstract: A graphic abstract is available for this article.
Authors: Seema Jain; Wesley H Self; Richard G Wunderink; Sherene Fakhran; Robert Balk; Anna M Bramley; Carrie Reed; Carlos G Grijalva; Evan J Anderson; D Mark Courtney; James D Chappell; Chao Qi; Eric M Hart; Frank Carroll; Christopher Trabue; Helen K Donnelly; Derek J Williams; Yuwei Zhu; Sandra R Arnold; Krow Ampofo; Grant W Waterer; Min Levine; Stephen Lindstrom; Jonas M Winchell; Jacqueline M Katz; Dean Erdman; Eileen Schneider; Lauri A Hicks; Jonathan A McCullers; Andrew T Pavia; Kathryn M Edwards; Lyn Finelli Journal: N Engl J Med Date: 2015-07-14 Impact factor: 91.245
Authors: Richard Y-C Huang; James G Laing; Evelyn M Kanter; Viviana M Berthoud; Mingwei Bao; Henry W Rohrs; R Reid Townsend; Kathryn A Yamada Journal: J Proteome Res Date: 2011-02-04 Impact factor: 4.466
Authors: Mari Numata; James R Mitchell; Jennifer L Tipper; Jeffrey D Brand; John E Trombley; Yoji Nagashima; Pitchaimani Kandasamy; Hong Wei Chu; Kevin S Harrod; Dennis R Voelker Journal: J Biol Chem Date: 2019-12-27 Impact factor: 5.157
Authors: Melissa A Rolfes; Ivo M Foppa; Shikha Garg; Brendan Flannery; Lynnette Brammer; James A Singleton; Erin Burns; Daniel Jernigan; Sonja J Olsen; Joseph Bresee; Carrie Reed Journal: Influenza Other Respir Viruses Date: 2018-02-14 Impact factor: 4.380
Authors: Helen T Groves; Jacqueline U McDonald; Pinky Langat; Ekaterina Kinnear; Paul Kellam; John McCauley; Joanna Ellis; Catherine Thompson; Ruth Elderfield; Lauren Parker; Wendy Barclay; John S Tregoning Journal: Front Immunol Date: 2018-01-31 Impact factor: 7.561