Role of matrix metalloproteinases in the pathogenesis of idiopathic pulmonary fibrosis

Annie Pardo, Sandra Cabrera, Mariel Maldonado, Moisés Selman, Annie Pardo, Sandra Cabrera, Mariel Maldonado, Moisés Selman

Abstract

Idiopathic pulmonary fibrosis (IPF) is a progressive and devastating lung disorder of unknown origin, with very poor prognosis and no effective treatment. The disease is characterized by abnormal activation of alveolar epithelial cells, which secrete numerous mediators involved in the expansion of the fibroblast population, its differentiation to myofibroblasts, and in the exaggerated accumulation of extracellular matrix provoking the loss of lung architecture. Among the excessively produced mediators are several matrix metalloproteases (MMPs) which may contribute to modify the lung microenvironment by various mechanisms. Thus, these enzymes can not only degrade all the components of the extracellular matrix, but they are also able to release, cleave and activate a wide range of growth factors, cytokines, chemokines and cell surface receptors affecting numerous cell functions including adhesion, proliferation, differentiation, recruiting and transmigration, and apoptosis. Therefore, dysregulated expression of MMPs may have profound impact on the biopathological mechanisms implicated in the development of IPF. This review focuses on the current and emerging evidence regarding the role of MMPs on the fibrotic processes in IPF as well as in mouse models of lung fibrosis.

Figures

Fig. 1
Fig. 1
Putative roles of some MMPs in pulmonary fibrosis. In activated alveolar epithelial cells, MMP-7 cleaves osteopontin and potentiates its function which in turn upregulates and activates MMP-7. In this profibrotic cross-talk, osteopontin upregulates the expression of ECM proteins. MMP-19 co-localizes and co-regulates with COX2 which end product, prostaglandin E2, is a potent suppressor of fibroblast proliferation and collagen production. Deficiency of MMP-19 in fibroblasts results in the upregulation of several profibrotic genes and pathways that provoke an increase of fibroblast migration and proliferation and a decrease of these processes in epithelial cells. Upregulation of MMP-1 in alveolar epithelial cells represses mitochondrial respiration and oxidative stress, while promotes cell proliferation, migration, and HIF-1α expression, and induces an anti-apoptotic phenotype. MMP3 induces EMT, and MMP9 is upregulated by TGF β in Thy-1 negative fibroblasts which in turn activates this growth factor. OPN = osteopontin; COX2 = cyclooxygenase 2; ECM = extracellular matrix; EMT = epithelial to mesenchymal transition

References

    1. King TE, Jr, Pardo A, Selman M. Idiopathic pulmonary fibrosis. Lancet. 2011;378:1949–61. doi: 10.1016/S0140-6736(11)60052-4.
    1. Raghu G, Collard HR, Egan JJ, Martinez FJ, Behr J, Brown KK, et al. An official ATS/ERS/JRS/ALAT statement: idiopathic pulmonary fibrosis: evidence-based guidelines for diagnosis and management. Am J Respir Crit Care Med. 2011;183:788–824. doi: 10.1164/rccm.2009-040GL.
    1. Thannickal VJ, Murthy M, Balch WE, Chandel NS, Meiners S, Eickelberg O, et al. Blue journal conference. Aging and susceptibility to lung disease. Am J Respir Crit Care Med. 2015;191:261–9. doi: 10.1164/rccm.201410-1876PP.
    1. Selman M, Pardo A. Revealing the pathogenic and aging-related mechanisms of the enigmatic idiopathic pulmonary fibrosis. An integral model. Am J Respir Crit Care Med. 2014;189:1161–72. doi: 10.1164/rccm.201312-2221PP.
    1. Yang IV, Pedersen BS, Rabinovich E, Hennessy CE, Davidson EJ, Murphy E, et al. Relationship of DNA methylation and gene expression in idiopathic pulmonary fibrosis. Am J Respir Crit Care Med. 2014;190:1263–72. doi: 10.1164/rccm.201408-1452OC.
    1. Selman M, Pardo A. Role of epithelial cells in idiopathic pulmonary fibrosis: from innocent targets to serial killers. Proc Am Thorac Soc. 2006;3:364–72. doi: 10.1513/pats.200601-003TK.
    1. Pardo A, Selman M, Kaminski N. Approaching the degradome in idiopathic pulmonary fibrosis. Int J Biochem Cell Biol. 2008;40:1141–55. doi: 10.1016/j.biocel.2007.11.020.
    1. Bonnans C, Chou J, Werb Z. Remodelling the extracellular matrix in development and disease. Nat Rev Mol Cell Biol. 2014;15:786–801. doi: 10.1038/nrm3904.
    1. Hynes RO, Naba A. Overview of the matrisome--an inventory of extracellular matrix constituents and functions. Cold Spring Harb Perspect Biol. 2012;4:a004903. doi: 10.1101/cshperspect.a004903.
    1. Puente XS, Sanchez LM, Overall CM, Lopez-Otin C. Human and mouse proteases: a comparative genomic approach. Nat Rev Genet. 2003;4:544–58. doi: 10.1038/nrg1111.
    1. Gaffney J, Solomonov I, Zehorai E, Sagi I. Multilevel regulation of matrix metalloproteinases in tissue homeostasis indicates their molecular specificity in vivo. Matrix Biol. 2015;44–46:191–9. doi: 10.1016/j.matbio.2015.01.012.
    1. Chou J, Shahi P, Werb Z. microRNA-mediated regulation of the tumor microenvironment. Cell Cycle. 2013;12:3262–71. doi: 10.4161/cc.26087.
    1. Limb GA, Matter K, Murphy G, Cambrey AD, Bishop PN, Morris GE, Khaw PT. Matrix metalloproteinase-1 associates with intracellular organelles and confers resistance to lamin A/C degradation during apoptosis. Am J Pathol. 2005;166:1555–63. doi: 10.1016/S0002-9440(10)62371-1.
    1. Wang W, Schulze CJ, Suarez-Pinzon WL, Sawicka J, Sawicki G, Schulz R. Intracellular action of matrix metalloproteinase-2 accounts for acute myocardial ischemia and reperfusion injury. Circulation. 2002;106:1543–9. doi: 10.1161/01.CIR.0000028818.33488.7B.
    1. Eguchi T, Kubota S, Kawata K, Mukudai Y, Uehara J, Ohgawara T, et al. Novel transcription-factor-like function of human matrix metalloproteinase 3 regulating the CTGF⁄CCN2 gene. Mol Cell Biol. 2008;28:2391–413. doi: 10.1128/MCB.01288-07.
    1. Cauwe B, Martens E, Proost P, Opdenakker G. Multidimensional degradomics identifies systemic autoantigens and intracellular matrix proteins as novel gelatinase B/MMP-9 substrates. Integr Biol (Camb) 2009;1:404–26. doi: 10.1039/b904701h.
    1. Shimizu-Hirota R, Xiong W, Baxter BT, Kunkel SL, Maillard I, Chen XW, et al. MT1-MMP regulates the PI3Kδ•Mi-2/NuRD-dependent control of macrophage immune function. Genes Dev. 2012;26:395–413. doi: 10.1101/gad.178749.111.
    1. Marchant DJ, Bellac CL, Moraes TJ, Wadsworth SJ, Dufour A, Butler GS, et al. A new transcriptional role for matrix metalloproteinase-12 in antiviral immunity. Nature Med. 2014;20:493–502. doi: 10.1038/nm.3508.
    1. Zuo F, Kaminski N, Eugui E, Allard J, Yakhini Z, Ben-Dor A, et al. Gene expression analysis reveals matrilysin as a key regulator of pulmonary fibrosis in mice and humans. Proc Natl Acad Sci U S A. 2002;99:6292–7. doi: 10.1073/pnas.092134099.
    1. Selman M, Pardo A, Barrera L, Estrada A, Watson SR, Wilson K, et al. Gene expression profiles distinguish idiopathic pulmonary fibrosis from hypersensitivity pneumonitis. Am J Respir Crit Care Med. 2006;173:188–98. doi: 10.1164/rccm.200504-644OC.
    1. Morrison CJ, Butler GS, Rodríguez D, Overall CM. Matrix metalloproteinase proteomics: substrates, targets, and therapy. Curr Opin Cell Biol. 2009;21:645–53. doi: 10.1016/j.ceb.2009.06.006.
    1. Pardo A, Gibson K, Cisneros J, Richards TJ, Yang Y, Becerril C, et al. Up-regulation and profibrotic role of osteopontin in human idiopathic pulmonary fibrosis. PLoS Med. 2005;2 doi: 10.1371/journal.pmed.0020251.
    1. Agnihotri R, Crawford HC, Haro H, Matrisian LM, Havrda MC, Liaw L. Osteopontin, a novel substrate for matrix metalloproteinase-3[stromelysin-1] and matrix metalloproteinase-7 [matrilysin] J Biol Chem. 2001;276:28261–7. doi: 10.1074/jbc.M103608200.
    1. Yu G, Kovkarova-Naumovski E, Jara P, Parwani A, Kass D, Ruiz V, et al. Matrix metalloproteinase-19 is a key regulator of lung fibrosis in mice and humans. Am J Respir Crit Care Med. 2012;186:752–62. doi: 10.1164/rccm.201202-0302OC.
    1. Hodges RJ, Jenkins RG, Wheeler-Jones CP, Copeman DM, Bottoms SE, Bellingan GJ, et al. Severity of lung injury in cyclooxygenase-2-deficient mice is dependent on reduced prostaglandin E(2) production. Am J Pathol. 2004;165:1663–76. doi: 10.1016/S0002-9440(10)63423-2.
    1. Jara P, Calyeca J, Romero Y, Plácido L, Yu G, Kaminski N, et al. Matrix metalloproteinase (MMP)-19-deficient fibroblasts display a profibrotic phenotype. Am J Physiol Lung Cell Mol Physiol. 2015;308:L511–22. doi: 10.1152/ajplung.00043.2014.
    1. Selman M, Ruiz V, Cabrera S, Segura L, Ramírez R, Barrios R, Pardo A. TIMP-1, −2, −3 and −4 in idiopathic pulmonary fibrosis. A prevailing non degradative lung microenvironment? Am J Physiol. 2000;279:L562–74.
    1. Segura-Valdez L, Pardo A, Gaxiola M, Uhal BD, Becerril C, Selman M. Upregulation of gelatinases A and B, collagenases 1 and 2, and increased parenchymal cell death in COPD. Chest. 2000;117:684–94. doi: 10.1378/chest.117.3.684.
    1. Vincenti MP, Brinckerhoff CE. Transcriptional regulation of collagenase [MMP-1, MMP-13] genes in arthritis: integration of complex signaling pathways for the recruitment of gene-specific transcription factors. Arthritis Res. 2002;4:157–64. doi: 10.1186/ar401.
    1. Imai K, Dalal SS, Chen ES, Downey R, Schulman LL, Ginsburg M, D’Armiento J. Human collagenase (matrix metalloproteinase-1) expression in the lungs of patients with emphysema. Am J Respir Crit Care Med. 2001;163:786–91. doi: 10.1164/ajrccm.163.3.2001073.
    1. Mercer BA, Kolesnikova N, Sonett J, D’Armiento J. Extracellular regulated kinase/mitogen-activated protein kinase is up-regulated in pulmonary emphysema and mediates matrix metalloproteinase-1 induction by cigarette smoke. J Biol Chem. 2004;279:17690–6. doi: 10.1074/jbc.M313842200.
    1. Herrera I, Cisneros J, Maldonado M, Ramírez R, Ortiz-Quintero B, Anso E, et al. Matrix metalloproteinase (MMP)-1 induces lung alveolar epithelial cell migration and proliferation, protects from apoptosis, and represses mitochondrial oxygen consumption. J Biol Chem. 2013;288:25964–75. doi: 10.1074/jbc.M113.459784.
    1. Balbín M, Fueyo A, Knäuper V, López JM, Alvarez J, Sánchez LM, et al. Identification and enzymatic characterization of two diverging murine counterparts of human interstitial collagenase (MMP-1) expressed at sites of embryo implantation. J Biol Chem. 2001;276:10253–62. doi: 10.1074/jbc.M009586200.
    1. Nuttall RK, Sampieri CL, Pennington CJ, Gill SE, Schultz GA, Edwards DR. Expression analysis of the entire MMP and TIMP gene families during mouse tissue development. FEBS Lett. 2004;563:129–34. doi: 10.1016/S0014-5793(04)00281-9.
    1. Yamashita CM, Dolgonos L, Zemans RL, Young SK, Robertson J, Briones N, et al. Matrix metalloproteinase 3 is a mediator of pulmonary fibrosis. Am J Pathol. 2011;179:1733–45. doi: 10.1016/j.ajpath.2011.06.041.
    1. Kaminski N, Allard JD, Pittet JF, Zuo F, Griffiths MJ, Morris D, et al. Global analysis of gene expression in pulmonary fibrosis reveals distinct programs regulating lung inflammation and fibrosis. Proc Natl Acad Sci U S A. 2000;97:1778–83. doi: 10.1073/pnas.97.4.1778.
    1. Cabrera S, Selman M, Lozano-Bolaños A, Konishi K, Richards TJ, Kaminski N, Pardo A. Gene expression profiles reveal molecular mechanisms involved in the progression and resolution of bleomycin-induced lung fibrosis. Am J Physiol Lung Cell Mol Physiol. 2013;304:L593–601. doi: 10.1152/ajplung.00320.2012.
    1. Hagood JS, Prabhakaran P, Kumbla P, Salazar L, MacEwen MW, Barker TH, et al. Loss of fibroblast Thy-1 expression correlates with lung fibrogenesis. Am J Pathol. 2005;167:365–79. doi: 10.1016/S0002-9440(10)62982-3.
    1. Ramírez G, Hagood JS, Sanders Y, Ramírez R, Becerril C, Segura L, et al. Absence of Thy-1 results in TGF-β induced MMP-9 expression and confers a profibrotic phenotype to human lung fibroblasts. Lab Invest. 2011;91:1206–18. doi: 10.1038/labinvest.2011.80.
    1. Yu Q, Stamenkovic I. Cell surface-localized matrix metalloproteinase-9 proteolytically activates TGF-β and promotes tumor invasion and angiogenesis. Genes Dev. 2000;14:163–76.
    1. Betsuyaku T, Fukuda Y, Parks WC, Shipley JM, Senior RM. Gelatinase B is required for alveolar bronchiolization after intratracheal bleomycin. Am J Pathol. 2000;157:525–35. doi: 10.1016/S0002-9440(10)64563-4.
    1. Cabrera S, Gaxiola M, Arreola JL, Ramírez R, Jara P, D’Armiento J, et al. Overexpression of MMP9 in macrophages attenuates pulmonary fibrosis induced by bleomycin. Int J Biochem Cell Biol. 2007;39:2324–38. doi: 10.1016/j.biocel.2007.06.022.
    1. Bucala R. Review Series--Inflammatio & Fibrosis. Fibrocytes and fibrosis QJM. 2012;105:505–8.
    1. Moeller A, Gilpin SE, Ask K, Cox G, Cook D, Gauldie J, et al. Circulating fibrocytes are an indicator of poor prognosis in idiopathic pulmonary fibrosis. Am J Respir Crit Care Med. 2009;179:588–94. doi: 10.1164/rccm.200810-1534OC.
    1. Andersson-Sjöland A, de Alba CG, Nihlberg K, Becerril C, Ramírez R, Pardo A, et al. Fibrocytes are a potential source of lung fibroblasts in idiopathic pulmonary fibrosis. Int J Biochem Cell Biol. 2008;40:2129–40. doi: 10.1016/j.biocel.2008.02.012.
    1. García-de-Alba C, Becerril C, Ruiz V, González Y, Reyes S, García-Alvarez J, Selman M, Pardo A. Expression of matrix metalloproteases by fibrocytes: possible role in migration and homing. Am J Respir Crit Care Med. 2010;182:1144–52. doi: 10.1164/rccm.201001-0028OC.
    1. Rosas IO, Richards TJ, Konishi K, Zhang Y, Gibson K, Lokshin AE, et al. MMP1 and MMP7 as potential peripheral blood biomarkers in idiopathic pulmonary fibrosis. PLoS Med. 2008;5 doi: 10.1371/journal.pmed.0050093.
    1. McKeown S, Richter AG, O’Kane C, McAuley DF, Thickett DR. MMP expression and abnormal lung permeability are important determinants of outcome in IPF. Eur Respir J. 2009;33:77–84. doi: 10.1183/09031936.00060708.
    1. Willems S, Verleden SE, Vanaudenaerde BM, Wynants M, Dooms C, Yserbyt J, et al. Multiplex protein profiling of bronchoalveolar lavage in idiopathic pulmonary fibrosis and hypersensitivity pneumonitis. Ann Thorac Med. 2013;8:38–45. doi: 10.4103/1817-1737.105718.
    1. Craig VJ, Polverino F, Laucho-Contreras ME, Shi Y, Liu Y, Osorio JC, et al. Mononuclear phagocytes and airway epithelial cells: novel sources of matrix metalloproteinase-8 (MMP-8) in patients with idiopathic pulmonary fibrosis. PLoS ONE. 2014;9 doi: 10.1371/journal.pone.0097485.
    1. Craig VJ, Quintero PA, Fyfe SE, Patel AS, Knolle MD, Kobzik L, Owen CA. Profibrotic activities for matrix metalloproteinase-8 during bleomycin-mediated lung injury. J Immunol. 2013;190:4283–96. doi: 10.4049/jimmunol.1201043.
    1. García-Prieto E, González-López A, Cabrera S, Astudillo A, Gutiérrez-Fernández A, Fanjul-Fernandez M, et al. Resistance to bleomycin-induced lung fibrosis in MMP-8 deficient mice is mediated by interleukin-10. PLoS ONE. 2010;5 doi: 10.1371/journal.pone.0013242.
    1. Pardo A, Barrios R, Gaxiola M, Segura-Valdez L, Carrillo G, Estrada A, et al. Increase of lung neutrophils in hypersensitivity pneumonitis is associated with lung fibrosis. Am J Respir Crit Care Med. 2000;161:1698–704. doi: 10.1164/ajrccm.161.5.9907065.
    1. Nkyimbeng T, Ruppert C, Shiomi T, Dahal B, Lang G, Seeger W, et al. Pivotal role of matrix metalloproteinase 13 in extracellular matrix turnover in idiopathic pulmonary fibrosis. PLoS ONE. 2013;8(9) doi: 10.1371/journal.pone.0073279.
    1. Flechsig P, Hartenstein B, Teurich S, Dadrich M, Hauser K, Abdollahi A, et al. Loss of matrix metalloproteinase-13 attenuates murine radiation-induced pulmonary fibrosis. Int J Radiat Oncol Biol Phys. 2010;77:582–90. doi: 10.1016/j.ijrobp.2009.12.043.
    1. Sen AI, Shiomi T, Okada Y, D’Armiento JM. Deficiency of matrix metalloproteinase-13 increases inflammation after acute lung injury. Exp Lung Res. 2010;36:615–24. doi: 10.3109/01902148.2010.497201.
    1. Manoury B, Nenan S, Guenon I, Boichot E, Planquois JM, Bertrand CP, Lagente V. Macrophage metalloelastase (MMP-12) deficiency does not alter bleomycin-induced pulmonary fibrosis in mice. J Inflamm (Lond) 2006;3:2. doi: 10.1186/1476-9255-3-2.
    1. Kang HR, Cho SJ, Lee CG, Homer RJ, Elias JA. Transforming growth factor (TGF)-beta1 stimulates pulmonary fibrosis and inflammation via a Bax-dependent, bid-activated pathway that involves matrix metalloproteinase-12. J Biol Chem. 2007;282:7723–32. doi: 10.1074/jbc.M610764200.
    1. Matute-Bello G, Wurfel MM, Lee JS, Park DR, Frevert CW, Madtes DK, et al. Essential role of MMP-12 in Fas-induced lung fibrosis. Am J Respir Cell Mol Biol. 2007;37:210–21. doi: 10.1165/rcmb.2006-0471OC.
    1. Peng R, Sridhar S, Tyagi G, Phillips JE, Garrido R, Harris P, et al. Bleomycin induces molecular changes directly relevant to idiopathic pulmonary fibrosis: a model for “active” disease. PLoS ONE. 2013;8 doi: 10.1371/journal.pone.0059348.
    1. Sabeh F, Li XY, Saunders TL, Rowe RG, Weiss SJ. Secreted versus membrane-anchored collagenases: relative roles in fibroblast-dependent collagenolysis and invasion. J Biol Chem. 2009;284:23001–11. doi: 10.1074/jbc.M109.002808.
    1. Rowe RG, Keena D, Sabeh F, Willis AL, Weiss SJ. Pulmonary fibroblasts mobilize the membrane-tethered matrix metalloprotease, MT1-MMP, to destructively remodel and invade interstitial type I collagen barriers. Am J Physiol Lung Cell Mol Physiol. 2011;301:L683–92. doi: 10.1152/ajplung.00187.2011.
    1. García-Alvarez J, Ramirez R, Sampieri CL, Nuttall RK, Edwards DR, Selman M, Pardo A. Membrane type-matrix metalloproteinases in idiopathic pulmonary fibrosis. Sarcoidosis Vasc Diffuse Lung Dis. 2006;23:13–21.
    1. Gharib SA, Johnston LK, Huizar I, Birkland TP, Hanson J, Wang Y, et al. MMP28 promotes macrophage polarization toward M2 cells and augments pulmonary fibrosis. J Leukoc Biol. 2014;95:9–18. doi: 10.1189/jlb.1112587.
    1. Ulivi P, Casoni GL, Foschi G, Scarpi E, Tomassetti S, Romagnoli M, et al. MMP-7 and fcDNA serum levels in early NSCLC and idiopathic interstitial pneumonia: preliminary study. Int J Mol Sci. 2013;14:24097–112. doi: 10.3390/ijms141224097.
    1. Morais A, Beltrão M, Sokhatska O, Costa D, Melo N, Mota P, et al. Serum metalloproteinases 1 and 7 in the diagnosis of idiopathic pulmonary fibrosis and other interstitial pneumonias. Respir Med. 2015;109:1063–8. doi: 10.1016/j.rmed.2015.06.003.
    1. Sanders YY, Ambalavanan N, Halloran B, Zhang X, Liu H, Crossman DK, et al. Altered DNA methylation profile in idiopathic pulmonary fibrosis. Am J Respir Crit Care Med. 2012;186:525–35. doi: 10.1164/rccm.201201-0077OC.
    1. Richards TJ, Kaminski N, Baribaud F, Flavin S, Brodmerkel C, Horowitz D, et al. Peripheral blood proteins predict mortality in idiopathic pulmonary fibrosis. Am J Respir Crit Care Med. 2012;185:67–76. doi: 10.1164/rccm.201101-0058OC.
    1. Checa M, Ruiz V, Montaño M, Velázquez-Cruz R, Selman M, Pardo A. MMP-1 polymorphisms and the risk of idiopathic pulmonary fibrosis. Hum Genet. 2008;124:465–72. doi: 10.1007/s00439-008-0571-z.
    1. DePianto DJ, Chandriani S, Abbas AR, Jia G, N’Diaye EN, Caplazi P, et al. Heterogeneous gene expression signatures correspond to distinct lung pathologies and biomarkers of disease severity in idiopathic pulmonary fibrosis. Thorax. 2015;70:48–56. doi: 10.1136/thoraxjnl-2013-204596.
    1. Sokai A, Handa T, Tanizawa K, Oga T, Uno K, Tsuruyama T, et al. Matrix metalloproteinase-10: a novel biomarker for idiopathic pulmonary fibrosis. Respir Res. 2015;16:120. doi: 10.1186/s12931-015-0280-9.
    1. Jenkins RG, Simpson JK, Saini G, Bentley JH, Russell AM, Braybrooke R, et al. Longitudinal change in collagen degradation biomarkers in idiopathic pulmonary fibrosis: an analysis from the prospective, multicentre PROFILE study. Lancet Respir Med. 2015;3:462–72. doi: 10.1016/S2213-2600(15)00048-X.
    1. Cathcart J, Pulkoski-Gross A, Cao J. Targeting Matrix Metalloproteinases in Cancer: Bringing New Life to Old Ideas. Genes Dis. 2015;2:26–34. doi: 10.1016/j.gendis.2014.12.002.
    1. Holmbeck K, Bianco P, Caterina J, Yamada S, Kromer M, Kuznetsov SA, et al. MT1-MMP-deficient mice develop dwarfism, osteopenia, arthritis, and connective tissue disease due to inadequate collagen turnover. Cell. 1999;99:81–92. doi: 10.1016/S0092-8674(00)80064-1.
    1. Manicone AM, Harju-Baker S, Johnston LK, Chen AJ, Parks WC. Epilysin (Matrix Metalloproteinase-28) Contributes to Airway Epithelial Cell Survival. Respir Res. 2011;12:144. doi: 10.1186/1465-9921-12-144.

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