[en] The term angiogenesis refers to sprouting of new blood vessels from pre-existing ones. The angiogenic process involves cell migration and tubulogenesis requiring interaction between endothelial cells and the extracellular ma- trix. Human peroxidasin 1 (hsPxd01) is a multidomain heme peroxidase found embedded in the basement mem- branes. As it promotes the stabilization of extracellular matrix, we investigated its possible role in angiogenesis
both in vitro and in vivo.
We analysed the effects of peroxidasin 1 gene silencing and supplementation by recombinant hsPxd01 in
TeloHAEC endothelial cells on cell migration, tubulogenesis in matrigel, and intracellular signal transduction as
assessed by kinase phosphorylation and expression of pro-angiogenic genes as measured by qRT–PCR. We further
evaluated the angiogenic potential of recombinant peroxidasin in a chicken chorioallantoic membrane model. RNA
silencing of endogenous hsPxd01 significantly reduced tube formation and cell migration, whereas supplementation
by the recombinant peroxidase promoted tube formation in vitro and stimulated vascularization in vivo through its
catalytic activity. Moreover, recombinant hsPxd01 promoted phosphorylation of Extracellular signal-Regulated
Kinases (ERK1/2), Protein kinase B (Akt), and Focal Adhesion Kinase (FAK), and induced the expression of pro-
angiogenic downstream genes: Platelet Derived Growth Factor Subunit B (PDGFB), endothelial-derived Heparin
Binding EGF-like growth factor (HB-EGF), CXCL-1, Hairy-Related Transcription Factor 1 (HEY-1), DNA-binding
protein inhibitor (ID-2), Snail Family Zinc Finger 1 (SNAI-1), as well as endogenous hsPxd01. However, peroxidasin
silencing significantly reduced Akt and FAK phosphorylation but induced ERK1/2 activation after supplementation
by recombinant hsPxd01. While hsPxd01 silencing significantly reduced expression of HEY-1, ID-2, and PDGFB, it
did not affect expression of SNAI-1, HB-EGF, and CXCL-1 after supplementation by recombinant hsPxd01.
Our findings suggest a role of enzymatically active peroxidasin 1 as a pro-angiogenic peroxidase and a modulator of ERK1/2, Akt and FAK signalling.
Disciplines :
Oncology
Author, co-author :
Medfai, Hayfa
Khalil, Alia
Rousseau, Alexandre
Nuyens, Vincent
Paumann-Page, Martina
Sevcnikar, Benjamin
Furtmüller, Paul
Obinger, Christian
Moguilevsky, Nicole
Peulen, Olivier ; Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Département des sciences biomédicales et précliniques
Herfs, Michael ; Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Anatomie et cytologie pathologiques
Castronovo, Vincenzo ; Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Biologie générale et cellulaire
Ucuzian AA, Gassman AA, East AT, Greisler HP. Molecular mediators of angiogenesis. J Burn Care Res 2010; 31: 158-175.
Kumar R, Yoneda J, Bucana CD, Fidler IJ. Regulation of distinct steps of angiogenesis by different angiogenic molecules. Int J Oncol 1998; 12: 749-757.
De Cicco-Skinner KL, Henry GH, Cataisson C, Tabib T, Gwilliam JC, Watson NJ, Bullwinkle EM, Falkenburg L, O'Neill RC, Morin A, Wiest JS. Endothelial cell tube formation assay for the in vitro study of angiogenesis. J Vis Exp 2014; 91: e51312.
Grant DS, Kibbey MC, Kinsella JL, Cid MC, Kleinman HK. The role of basement membrane in angiogenesis and tumor growth. Pathol Res Pract 1994; 190: 854-863.
Kim WY, Lee HY. Brain angiogenesis: mechanism and therapeutic intervention in brain tumors. FEBS J 2009; 276: 4653-4664.
Liu Y, Senger DR. Matrix-specific activation of Src and Rho initiates capillary morphogenesis of endothelial cells. FASEB J 2004; 18: 457-468.
Senger DR, Claffey KP, Benes JE, Perruzzi CA, Sergiou AP, Detmar M. Angiogenesis promoted by vascular endothelial growth factor: regulation through alpha1beta1 and alpha2beta1 integrins. Proc Natl Acad Sci USA 1997; 94: 13612-13617.
Lo C-M, Wang H-B, Dembo M, Wang Y-L. Cell movement is guided by the rigidity of the substrate. Biophys J 2000; 79: 144-152.
Hoeben A, Landuyt B, Highley MS, Wildiers H, Van Oosterom AT, De Bruijn EA. Vascular endothelial growth factor and angiogenesis. Pharmacol Rev 2004; 56: 549-580.
Carmeliet P. Angiogenesis in life, disease and medicine. Nature 2005; 438: 932-936.
Cheng G, Li H, Cao Z, Qiu X, McCormick S, Thannickal VJ, Nauseef WM. Vascular Peroxidase-1 is rapidly secreted circulates in plasma, and supports dityrosine crosslinking reactions. Free Radic Biol Med 2011; 51: 1445-1453.
Zhang YS, He L, Liu B, Li NS, Luo XJ, Hu CP, Ma QL, Zhang GG, Li YJ, Peng J. A novel pathway of NADPH oxidase/vascular peroxidase 1 in mediating oxidative injury following ischemia-reperfusion. Basic Res Cardiol 2012; 107: 266.
Shi R, Hu C, Yuan Q, Yang T, Peng J, Li Y, Bai Y, Cao Z, Cheng G, Zhang G. Involvement of vascular peroxidase 1 in angiotensin II-induced vascular smooth muscle cell proliferation. Cardiovasc Res 2011; 91: 27-36.
Péterfi Z, Donkó A, Orient A, Sum A, Pró kai A, Molnár B, Veréb Z, Rajnavölgyi E, Kovács KJ, Müller V, Szabó AJ, Geiszt M. Peroxidasin is secreted and incorporated into the extracellular matrix of myofibroblasts and fibrotic kidney. Am J Pathol 2009; 175: 725-735.
Lázár E, Péterfi Z, Sirokmány G, Kovács HA, Klement E, Medzihradszky KF, Geiszt M. Structure-function analysis of peroxidasin provides insight into the mechanism of collagen IV crosslinking. Free Radic Biol Med 2015; 83: 273-282.
Soudi M, Zámocky M, Jakopitsch C, Furtmüller PG, Obinger C. Molecular evolution, structure, and function of peroxidasins. Chem Biodivers 2012; 9: 1776-1793.
Soudi M, Paumann-Page M, Delporte C, Pirker KF, Bellei M, Edenhofer E, Stadlmayr G, Battistuzzi G, Boudjeltia KZ, Furtmüller PG, Van Antwerpen P, Obinger C. Multidomain human peroxidasin 1 is a highly glycosylated and stable homotrimeric high spin ferric peroxidase. J Biol Chem 2015; 290: 10876-10890.
Cheng G, Salerno JC, Cao Z, Pagano PJ, Lambeth JD. Identification and characterization of VPO1, a new animal heme containing Peroxidase. Free Radic Biol Med 2008; 45: 1682-1694.
Nelson RE, Fessler LI, Takagi Y, Blumberg B, Keene DR, Olson PF, Parker CG, Fessler JH. Peroxidasin: a novel enzyme-matrix protein of Drosophila development. EMBO J 1994; 13: 3438-3447.
Bhave G, Cummings CF, Vanacore RM, Kumagai-Cresse C, Ero-Tolliver IA, Rafi M, Kang JS, Pedchenko V, Fessler LI, Fessler JH, Hudson BG. Peroxidasin forms sulfilimine chemical bonds using hypohalous acids in tissue genesis. Nat Chem Biol 2012; 8: 784-790.
Ero-Tolliver IA, Hudson BG, Bhave G. The ancient immunoglobulin domains of peroxidasin are required to form sulfilimine cross-links in collagen IV. J Biol Chem 2015; 290: 21741-21748.
Khan K, Rudkin A, Parry DA, Burdon KP, McKibbin M, Logan CV, Abdelhamed ZI, Muecke JS, Fernandez-Fuentes N, Laurie KJ, Shires M, Fogarty R, Carr IM, Poulter JA, Morgan JE, Mohamed MD, Jafri H, Raashid Y, Meng N, Piseth H, Toomes C, Casson RJ, Taylor GR, Hammerton M, Sheridan E, Johnson CA, Inglehearn CF, Craig JE, Ali M. Homozygous mutations in PXDN cause congenital cataract, corneal opacity, and developmental glaucoma. Am J Hum Genet 2011; 89: 464-473.
Liu Y, Carson-Walter EB, Cooper A, Winans BN, Johnson MD, Walter KA. Vascular gene expression patterns are conserved in primary and metastatic brain tumors. J Neurooncol 2010; 99: 13-24.
Castronovo V, Waltregny D, Kischel P, Roesli C, Elia G, Rybak JN, Neri D. A chemical proteomics approach for the identification of accessible antigens expressed in human kidney cancer. Mol Cell Proteomics 2006; 5: 2083-2091.
Van Hinsbergh VW, Koolwijk P. Endothelial sprouting and angiogenesis: matrix metalloproteinases in the lead. Cardiovasc Res 2008; 78: 203-212.
Neve A, Cantatore FP, Maruotti N, Corrado A, Ribatti D. Extracellular matrix modulates angiogenesis in physiological and pathological conditions. BioMed Res Int 2014; 2014: 1.
Herbert SP, Stainier DY. Molecular control of endothelial cell behaviour during blood vessel morphogenesis. Nat Rev Mol Cell Biol 2011; 12: 551-564.
Davis GE, Koh W, Stratman AN. Mechanisms controlling human endothelial lumen formation and tube assembly in three-dimensional extracellular matrices. Birth Defects Res C Embryo Today 2007; 81: 270-285.
Panagopoulos V, Zinonos I, Leach DA, Hay SJ, Liapis V, Zysk A, Ingman WV, DeNichilo MO, Evdokiou A. Uncovering a new role for peroxidase enzymes as drivers of angiogenesis. Int J Biochem Cell Biol 2015; 68: 128-138.
Zámocky M, Jakopitsch C, Furtmüller PG, Dunand C, Obinger C. The peroxidasecyclooxygenase superfamily: reconstructed evolution of critical enzymes of the innate immune system. Proteins 2008; 72: 589-605.
Zámocky M, Hofbauer S, Schaffner I, Gasselhuber B, Nicolussi A, Soudi M, Pirker KF, Furtmüller PG, Obinger C. Independent evolution of four heme peroxidase superfamilies. Arch Biochem Biophys 2015; 574: 108-119.
Longo PA, Kavran JM, Kim M-S, Leahy DJ. Transient mammalian cell transfection with polyethylenimine (PEI). Methods Enzymol 2013; 529: 227-240.
Zudaire E, Gambardella L, Kurcz C, Vermeren S. A computational tool for quantitative analysis of vascular networks. PLoS One 2011; 6: e27385.
Paumann-Page M, Katz RS, Bellei M, Schwartz I, Edenhofer E, Sevcnikar B, Soudi M, Hofbauer S, Battistuzzi G, Furtmüller PG, Obinger C. Pre-steady-state kinetics reveal the substrate specificity and mechanism of halide oxidation of truncated human peroxidasin 1. J Biol Chem 2017; 292: 4583-4592.
Colon S, Bhave G. Proprotein convertase processing enhances peroxidasin activity to reinforce collagen IV. J Biol Chem 2016; 291: 24009-24016.
Jayachandran A, Prithviraj P, Lo P-H, Walkiewicz M, Anaka M, Woods BL, Tan B, Behren A, Cebon J, McKeown SJ. Identifying and targeting determinants of melanoma cellular invasion. Oncotarget 2016; 7: 41186-41202.
Tauber S, Jais A, Jeitler M, Haider S, Husa J, Lindroos J, Knöfler M, Mayerhofer M, Pehamberger H, Wagner O, Bilban M. Transcriptome analysis of human cancer reveals a functional role of heme oxygenase-1 in tumor cell adhesion. Mol Cancer 2010; 9: 200.
Srinivasan R, Zabuawala T, Huang H, Zhang J, Gulati P, Fernandez S, Karlo JC, Landreth GE, Leone G, Ostrowski MC. Erk1 and Erk2 regulate endothelial cell proliferation and migration during mouse embryonic angiogenesis. PLoS One 2009; 4: e8283.
Tavora B, Batista S, Reynolds LE, Jadeja S, Robinson S, Kostourou V, Hart I, Fruttiger M, Parsons M, Hodivala-Dilke KM. Endothelial FAK is required for tumour angiogenesis. EMBO Mol Med 2010; 2: 516-528.
Jiang BH, Liu LZ. AKT signaling in regulating angiogenesis. Curr Cancer Drug Targets 2008; 8: 19-26.
Tang Y, Xu Q, Peng H, Liu Z, Yang T, Yu Z, Cheng G, Li X, Zhang G, Shi R. The role of vascular peroxidase 1 in ox-LDL-induced vascular smooth muscle cell calcification. Atherosclerosis 2015; 243: 357-363.
Li DJ, Zhao T, Xin RJ, Wang YY, Fei YB, Shen FM. Activation of a7 nicotinic acetylcholine receptor protects against oxidant stress damage through reducing vascular peroxidase-1 in a JNK signaling-dependent manner in endothelial cells. Cell Physiol Biochem 2014; 33: 468-478.
Bai YP, Hu CP, Yuan Q, Peng J, Shi RZ, Yang TL, Cao ZH, Li YJ, Cheng G, Zhang GG. Role of VPO1, a newly identified heme-containing peroxidase, in ox-LDL induced endothelial cell apoptosis. Free Radic Biol Med 2011; 51: 1492-1500.
Lau CK, Yang ZF, Ho DW, Ng MN, Yeoh GC, Poon RT, Fan ST. An Akt/hypoxiainducible factor-1 alpha/platelet-derived growth factor-BB autocrine loop mediates hypoxia-induced chemoresistance in liver cancer cells and tumorigenic hepatic progenitor cells. Clin Cancer Res 2009; 15: 3462-3471.
Bedogni B, Warneke JA, Nickoloff BJ, Giaccia AJ, Powell MB. Notch1 is an effector of Akt and hypoxia in melanoma development. J Clin Invest 2008; 118: 3660-3670.
Afrikanova I, Yebra M, Simpkinson M, Xu Y, Hayek A, Montgomery A. Inhibitors of Src and focal adhesion kinase promote endocrine specification: impact on the derivation of b-cells from human pluripotent stem cells. J Biol Chem 2011; 286: 36042-36052.
Baston-Büst DM, Schanz A, Böddeker SJ, Altergot-Ahmad O, Krüssel JS, Rein D, Hess AP. CXCL1 expression in human decidua in vitro is mediated via the MAPK signalling cascade. Cytokine 2013; 64: 79-85.
Kumasawa F, Hashimoto S, Mizumura K, Takeshita I, Onose A, Jibiki I, Maruoka S, Gon Y, Kobayashi T, Takahashi N. Mitogen-activated protein kinase (MAPK) regulates leukotriene D4-induced HB-EGF and ADAM12 expression in human airway smooth muscle cells. Asian Pac J Allergy Immunol 2013; 31: 58-66.
Wang S, Cheng Y, Zheng Y, He Z, Chen W, Zhou W, Duan C, Zhang C. PRKAR1A is a functional tumor suppressor inhibiting ERK/Snail/E-cadherin pathway in lung adenocarcinoma. Sci Rep 2016; 6: 39630.
Zhou Y, Yan H, Guo M, Zhu J, Xiao Q, Zhang L. Reactive oxygen species in vascular formation and development. Oxid Med Cell Longev 2013; 2013: 374963.
