Aging‐related Atg5 defect impairs neutrophil extracellular traps formation

Formation of neutrophil extracellular traps (NETs) is an important function of the innate immune system against infections. It has been proven that aging dysregulates immunity and impairs neutrophil function. However, the influence of aging on the ability to produce NETs has yet to be fully addressed. In this study, we tested the hypothesis that a lower level of autophagy in neutrophils from aged mice was responsible for the decrease in NET formation. We demonstrated that a broad range of Toll‐like receptor 2 (TLR2) ligands could efficiently induce reactive oxygen species (ROS) ‐dependent NET release in young mice, but not in aged ones. We further explored that the difference between young and aged mice in TLR2 ligand‐induced NETosis is the result of an Atg5 defect and subsequent impaired autophagy. Furthermore, we found that lower autophagy capacity led to not only reduced NET formation, but also increased apoptosis. Our results suggest an important role of Atg5 and autophagy in maintaining the function of NETs formation in response to infection and in regulating neutrophil death. Targeting autophagy‐promoted NETs may present a therapeutic strategy to improve infection defence in an aged population.

[1]  D. Bowdish,et al.  Infection in an aging population. , 2016, Current opinion in microbiology.

[2]  M. Machado,et al.  Septic Shock in Advanced Age: Transcriptome Analysis Reveals Altered Molecular Signatures in Neutrophil Granulocytes , 2015, PloS one.

[3]  W. Krautgartner,et al.  The Role of Reactive Oxygen Species (ROS) in the Formation of Extracellular Traps (ETs) in Humans , 2015, Biomolecules.

[4]  S. Yusuf,et al.  The burden of disease in older people and implications for health policy and practice , 2015, The Lancet.

[5]  Elizabeth Sapey,et al.  Impaired neutrophil extracellular trap formation: a novel defect in the innate immune system of aged individuals , 2014, Aging cell.

[6]  Eric H. Baehrecke,et al.  Self-consumption: the interplay of autophagy and apoptosis , 2014, Nature Reviews Molecular Cell Biology.

[7]  Peter E. Czabotar,et al.  Control of apoptosis by the BCL-2 protein family: implications for physiology and therapy , 2013, Nature Reviews Molecular Cell Biology.

[8]  Yoko Tabe,et al.  Neutrophil cell death in response to infection and its relation to coagulation , 2013, Journal of Intensive Care.

[9]  Ruth R. Montgomery,et al.  Age-dependent dysregulation of innate immunity , 2013, Nature Reviews Immunology.

[10]  O. McCarty,et al.  Pivotal role for the mTOR pathway in the formation of neutrophil extracellular traps via regulation of autophagy. , 2013, American journal of physiology. Cell physiology.

[11]  Wei-dong Hu,et al.  Application and interpretation of current autophagy inhibitors and activators , 2013, Acta Pharmacologica Sinica.

[12]  M. Czaja,et al.  Inhibition of hepatocyte autophagy increases tumor necrosis factor-dependent liver injury by promoting caspase-8 activation , 2013, Cell Death and Differentiation.

[13]  P. Rabinovitch,et al.  mTOR is a key modulator of ageing and age-related disease , 2013, Nature.

[14]  M. Asaduzzaman,et al.  Dynamic NETosis is Carried Out by Live Neutrophils in Human and Mouse Bacterial Abscesses and During Severe Gram-Positive Infection , 2012, Nature medicine.

[15]  M. Asaduzzaman,et al.  Infection-induced NETosis is a dynamic process involving neutrophil multitasking in vivo , 2012, Nature Medicine.

[16]  W. Tawackoli,et al.  Innate Immune Dysfunctions in Aged Mice Facilitate the Systemic Dissemination of Methicillin-Resistant S. aureus , 2012, PloS one.

[17]  D. Gozuacik,et al.  Cleavage of Atg3 protein by caspase-8 regulates autophagy during receptor-activated cell death , 2012, Apoptosis.

[18]  P. Massari,et al.  The Role of TLR2 in Infection and Immunity , 2012, Front. Immun..

[19]  S. Amin,et al.  Autophagosomal Membrane Serves as Platform for Intracellular Death-inducing Signaling Complex (iDISC)-mediated Caspase-8 Activation and Apoptosis* , 2012, The Journal of Biological Chemistry.

[20]  D. Rubinsztein,et al.  Autophagy and Aging , 2011, Cell.

[21]  D. Ganea,et al.  Cecal ligation puncture procedure. , 2011, Journal of visualized experiments : JoVE.

[22]  P. Vandenabeele,et al.  Dying for a cause: NETosis, mechanisms behind an antimicrobial cell death modality , 2011, Cell Death and Differentiation.

[23]  A. Reunanen,et al.  Aging reduces the functionality of anti-pneumococcal antibodies and the killing of Streptococcus pneumoniae by neutrophil phagocytosis. , 2011, Vaccine.

[24]  P. Vandenabeele,et al.  Neutrophil extracellular trap cell death requires both autophagy and superoxide generation , 2011, Cell Research.

[25]  K. Ryan,et al.  Androgens modulate autophagy and cell death via regulation of the endoplasmic reticulum chaperone glucose-regulated protein 78/BiP in prostate cancer cells , 2010, Cell Death and Disease.

[26]  R. Xavier,et al.  Genome-wide analysis reveals mechanisms modulating autophagy in normal brain aging and in Alzheimer's disease , 2010, Proceedings of the National Academy of Sciences.

[27]  R. Broaddus,et al.  The novel estrogen-induced gene EIG121 regulates autophagy and promotes cell survival under stress , 2010, Cell Death and Disease.

[28]  M. Lotz,et al.  Autophagy is a protective mechanism in normal cartilage, and its aging-related loss is linked with cell death and osteoarthritis. , 2010, Arthritis and rheumatism.

[29]  M. Sperandio,et al.  Delayed but functional neutrophil extracellular trap formation in neonates. , 2009, Blood.

[30]  D. Klionsky,et al.  Regulation mechanisms and signaling pathways of autophagy. , 2009, Annual review of genetics.

[31]  D. Rubinsztein,et al.  Apoptosis blocks Beclin 1-dependent autophagosome synthesis – an effect rescued by Bcl-xL , 2009, Cell Death and Differentiation.

[32]  Min-jung Park,et al.  Synergism of toll-like receptor 2 (TLR2), TLR4, and TLR6 ligation on the production of tumor necrosis factor (TNF)-alpha in a spontaneous arthritis animal model of interleukin (IL)-1 receptor antagonist-deficient mice. , 2009, Immunology letters.

[33]  A. Cuervo.  Autophagy and aging: keeping that old broom working. , 2008, Trends in genetics : TIG.

[34]  O. Lesur,et al.  Aging and neutrophils: there is still much to do. , 2008, Rejuvenation research.

[35]  F. Loison,et al.  Constitutive neutrophil apoptosis: Mechanisms and regulation , 2008, American journal of hematology.

[36]  V. Wahn,et al.  Novel cell death program leads to neutrophil extracellular traps. , 2007, The Journal of cell biology.

[37]  Sudhir Gupta,et al.  A paradox of immunodeficiency and inflammation in human aging: lessons learned from apoptosis , 2006, Immunity & Ageing.

[38]  J. El Benna,et al.  Inhibition of Neutrophil Apoptosis by TLR Agonists in Whole Blood: Involvement of the Phosphoinositide 3-Kinase/Akt and NF-κB Signaling Pathways, Leading to Increased Levels of Mcl-1, A1, and Phosphorylated Bad , 2005, The Journal of Immunology.

[39]  F. Finkelman,et al.  Bacterial Lipoprotein Delays Apoptosis in Human Neutrophils through Inhibition of Caspase-3 Activity: Regulatory Roles for CD14 and TLR-2 , 2004, The Journal of Immunology.

[40]  A. Zychlinsky,et al.  Neutrophil Extracellular Traps Kill Bacteria , 2004, Science.

[41]  A. Aderem,et al.  The repertoire for pattern recognition of pathogens by the innate immune system is defined by cooperation between toll-like receptors. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[42]  J. Stankova,et al.  Changes in apoptosis of human polymorphonuclear granulocytes with aging , 1997, Mechanisms of Ageing and Development.

[43]  M. Glogauer,et al.  Quantification and visualization of neutrophil extracellular traps (NETs) from murine bone marrow-derived neutrophils. , 2013, Methods in molecular biology.