Interferon regulatory factor 1–Rab27a regulated extracellular vesicles promote liver ischemia/reperfusion injury

The role and regulators of extracellular vesicle (EV) secretion in hepatic ischemia/reperfusion (IR) injury have not been defined. Rab27a is a guanosine triphosphatase known to control EV release. Interferon regulatory factor 1 (IRF‐1) is a transcription factor that plays an important role in liver IR and regulates certain guanosine triphosphatases. However, the relationships among IRF‐1, Rab27a, and EV secretion are largely unknown. Here, we show induction of IRF‐1 and Rab27a both in vitro in hypoxic hepatocytes and in vivo in warm IR and orthotopic liver transplantation livers. Interferon γ stimulation, IRF‐1 transduction, or IR promoted Rab27a expression and EV secretion. Meanwhile, silencing of IRF‐1 decreased Rab27a expression and EV secretion. Rab27a silencing decreased EV secretion and liver IR injury. Ten putative IRF‐1 binding motifs in the 1,692‐bp Rab27a promoter region were identified. Chromatin immunoprecipitation and electrophoretic mobility shift assay verified five functional IRF‐1 binding motifs, which were confirmed by a Rab27a promoter luciferase assay. IR‐induced EVs contained higher oxidized phospholipids (OxPL). OxPLs on the EV surface activated neutrophils through the toll‐like receptor 4 pathway. OxPL‐neutralizing E06 antibody blocked the effect of EVs and decreased liver IR injury. Conclusion: These findings provide a novel mechanism by which IRF‐1 regulates Rab27a transcription and EV secretion, leading to OxPL activation of neutrophils and subsequent hepatic IR injury. (Hepatology 2018;67:1056–1070)

[1]  B. Naini,et al.  Early cytokine signatures of ischemia/reperfusion injury in human orthotopic liver transplantation. , 2016, JCI insight.

[2]  James R. Springstead,et al.  An endogenous caspase-11 ligand elicits interleukin-1 release from living dendritic cells , 2016, Science.

[3]  C. Théry,et al.  Proteomic comparison defines novel markers to characterize heterogeneous populations of extracellular vesicle subtypes , 2016, Proceedings of the National Academy of Sciences.

[4]  Gary K. Schwartz,et al.  Tumour exosome integrins determine organotropic metastasis , 2015, Nature.

[5]  Christian Pilarsky,et al.  Glypican-1 identifies cancer exosomes and detects early pancreatic cancer , 2015, Nature.

[6]  S. Horvat,et al.  Toll-like receptor 4 senses oxidative stress mediated by the oxidation of phospholipids in extracellular vesicles , 2015, Science Signaling.

[7]  T. Wartmann,et al.  Lipidomic analysis of molecular cardiolipin species in livers exposed to ischemia/reperfusion , 2015, Molecular and Cellular Biochemistry.

[8]  V. Gladyshev,et al.  Endogenous Hydrogen Sulfide Production Is Essential for Dietary Restriction Benefits , 2015, Cell.

[9]  George A Calin,et al.  Cancer exosomes perform cell-independent microRNA biogenesis and promote tumorigenesis. , 2014, Cancer cell.

[10]  H. Ishwaran,et al.  Exosome Transfer from Stromal to Breast Cancer Cells Regulates Therapy Resistance Pathways , 2014, Cell.

[11]  T. Ørntoft,et al.  Cellular disposal of miR23b by RAB27-dependent exosome release is linked to acquisition of metastatic properties. , 2014, Cancer research.

[12]  T. Visakorpi,et al.  Different gDNA Content in the Subpopulations of Prostate Cancer Extracellular Vesicles: Apoptotic Bodies, Microvesicles, and Exosomes , 2014, The Prostate.

[13]  J. Rao,et al.  ATF6 Mediates a Pro‐Inflammatory Synergy Between ER Stress and TLR Activation in the Pathogenesis of Liver Ischemia‐Reperfusion Injury , 2014, American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons.

[14]  A. Falus,et al.  Emerging role of extracellular vesicles in inflammatory diseases , 2014, Nature Reviews Rheumatology.

[15]  P. Robbins,et al.  Regulation of immune responses by extracellular vesicles , 2014, Nature Reviews Immunology.

[16]  Vivek T. Natarajan,et al.  IFN-γ signaling maintains skin pigmentation homeostasis through regulation of melanosome maturation , 2014, Proceedings of the National Academy of Sciences.

[17]  P. Altevogt,et al.  Body Fluid Exosomes Promote Secretion of Inflammatory Cytokines in Monocytic Cells via Toll-like Receptor Signaling* , 2013, The Journal of Biological Chemistry.

[18]  Miharu Kobayashi,et al.  Hypoxia-Induced Changes in the Bioactivity of Cytotrophoblast-Derived Exosomes , 2013, PloS one.

[19]  J. Cleveland,et al.  Toll-Like Receptor 4–Dependent Microglial Activation Mediates Spinal Cord Ischemia–Reperfusion Injury , 2013, Circulation.

[20]  D. Stolz,et al.  Cellular‐specific role of toll‐like receptor 4 in hepatic ischemia‐reperfusion injury in mice , 2013, Hepatology.

[21]  Nuoyan Zheng,et al.  Guanylate Binding Protein 4 Negatively Regulates Virus-Induced Type I IFN and Antiviral Response by Targeting IFN Regulatory Factor 7 , 2011, The Journal of Immunology.

[22]  B. Binder,et al.  Oxidized Phospholipids Are More Potent Antagonists of Lipopolysaccharide than Inducers of Inflammation , 2010, The Journal of Immunology.

[23]  A. Tsung,et al.  Critical role of interferon regulatory factor‐1 in murine liver transplant ischemia reperfusion injury , 2010, Hepatology.

[24]  T. Tolmachova,et al.  Rab27a and Rab27b Regulate Neutrophil Azurophilic Granule Exocytosis and NADPH oxidase Activity by Independent Mechanisms , 2010, Traffic.

[25]  Sandra L. Schmid,et al.  Conserved Functions of Membrane Active GTPases in Coated Vesicle Formation , 2009, Science.

[26]  D. Vance,et al.  Phosphatidylcholine and choline homeostasis. , 2008, Journal of lipid research.

[27]  Arthur S Slutsky,et al.  Identification of Oxidative Stress and Toll-like Receptor 4 Signaling as a Key Pathway of Acute Lung Injury , 2008, Cell.

[28]  F. Gao,et al.  Type I, but not type II, interferon is critical in liver injury induced after ischemia and reperfusion , 2007, Hepatology.

[29]  S. Chadban,et al.  TLR4 activation mediates kidney ischemia/reperfusion injury. , 2007, The Journal of clinical investigation.

[30]  E. Brilakis,et al.  Oxidized phospholipids, Lp(a) lipoprotein, and coronary artery disease. , 2005, The New England journal of medicine.

[31]  J. Witztum,et al.  Apoptotic Cells with Oxidation-specific Epitopes Are Immunogenic and Proinflammatory , 2004, The Journal of experimental medicine.

[32]  J. MacMicking.  IFN-inducible GTPases and immunity to intracellular pathogens. , 2004, Trends in immunology.

[33]  Joseph Juliano,et al.  Temporal increases in plasma markers of oxidized low-density lipoprotein strongly reflect the presence of acute coronary syndromes. , 2003, Journal of the American College of Cardiology.

[34]  F. Gruber,et al.  Protective role of phospholipid oxidation products in endotoxin-induced tissue damage , 2002, Nature.

[35]  N. Lloberas,et al.  Postischemic renal oxidative stress induces an inflammatory response through PAF and oxidized phospholipids: prevention by antioxidant treatment , 2002, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[36]  E. Dennis,et al.  Correlation of Antiphospholipid Antibody Recognition with the Structure of Synthetic Oxidized Phospholipids , 2002, The Journal of Biological Chemistry.

[37]  B. Binder,et al.  Oxidized Membrane Vesicles and Blebs From Apoptotic Cells Contain Biologically Active Oxidized Phospholipids That Induce Monocyte‐Endothelial Interactions , 2002, Arteriosclerosis, thrombosis, and vascular biology.

[38]  K. Pfeffer,et al.  Two families of GTPases dominate the complex cellular response to IFN-gamma. , 1998, Journal of immunology.

[39]  R. Schuster,et al.  Hepatocyte exosomes mediate liver repair and regeneration via sphingosine-1-phosphate. , 2016, Journal of hepatology.

[40]  Miguel C. Seabra,et al.  Rab27a and Rab27b control different steps of the exosome secretion pathway , 2010, Nature Cell Biology.

[41]  T. Taniguchi,et al.  IRF family of transcription factors as regulators of host defense. , 2001, Annual review of immunology.