Advertisement

Endothelial cells of different organs exhibit heterogeneity in von Willebrand factor expression in response to hypoxia

      Highlights

      • von Willebrand factor (VWF) upregulation in response to hypoxia exhibits organ specificity.
      • Hypoxia-induced upregulation of VWF correlates with platelets aggregate formation.
      • Distinct molecular mechanisms regulate hypoxia-induced expression of VWF in heart compared to lung endothelial cells.

      Abstract

      Background and aims

      We have previously demonstrated that in response to hypoxia, von Willebrand factor (VWF) expression is upregulated in lung and heart endothelial cells both in vitro and in vivo, but not in kidney endothelial cells. The aim of our current study was to determine whether endothelial cells of different organs employ distinct molecular mechanisms to mediate VWF response to hypoxia.

      Methods

      We used cultured human primary lung, heart and kidney endothelial cells to determine the activation of endogenous VWF as well as exogenously expressed VWF promoter in response to hypoxia. Chromatin immunoprecipitation and siRNA knockdown analyses were used to determine the roles of VWF promoter associated transacting factors in mediating its hypoxia response. Platelet aggregates formations in vascular beds of mice were used as a marker for potential functional consequences of hypoxia-induced VWF upregulation in vivo.

      Results

      Our analyses demonstrated that while Yin Yang 1 (YY1) and specificity protein 1 (Sp1) participate in the hypoxia-induced upregulation of VWF specifically in lung endothelial cells, GATA6 mediates this process specifically in heart endothelial cells. In both cell types, the response to hypoxia involves the decreased association of the NFIB repressor with the VWF promoter, and the increased acetylation of the promoter-associated histone H4. In mice exposed to hypoxia, the upregulation of VWF expression was concomitant with the presence of thrombi in heart and lung, but not kidney vascular beds.

      Conclusions

      Heart and lung endothelial cells demonstrated VWF upregulation in response to hypoxia, using distinct mechanisms, while this response was lacking in kidney endothelial cells.

      Graphical abstract

      Keywords

      Abbreviations:

      ChIP (chromatin immunoprecipitation), HFF1 (human foreskin fibroblasts cell line), HPRT (hypoxanthine phosphoribosyltransferase), HIF-1α (hypoxia inducible factor 1α), IHC (immunohistochemistry), I51HSS (intron 51 hypersensitive sequence), MVEC (microvascular endothelial cells), MOI (multiplicity of infection), NFI (nuclear factor I), OCT (optimal cutting temperature compound), Sp1 (specificity protein 1), VWF (von Willebrand factor), YY1 (Yin Yang 1)
      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'

      Subscribe:

      Subscribe to Atherosclerosis
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect

      References

        • Springer T.A.
        • von
        Willebrand factor, Jedi knight of the bloodstream.
        Blood. 2014; 124: 1412-1425
        • Pusztaszeri M.P.
        • Seelentag W.
        • Bosman F.T.
        Immunohistochemical expression of endothelial markers CD31, CD34, von Willebrand factor, and Fli-1 in normal human tissues.
        J. Histochem. Cytochem. 2006; 54: 385-395
        • Yamamoto K.
        • de Waard V.
        • Fearns C.
        • Loskutoff D.J.
        Tissue distribution and regulation of murine von Willebrand factor gene expression in vivo.
        Blood. 1998; 92: 2791-2801
        • Aird W.C.
        Endothelial cell heterogeneity.
        Cold Spring Harbor Perspect. Med. 2012; 2: a006429
        • Mojiri A.
        • Nakhaii-Nejad M.
        • Phan W.L.
        • Kulak S.
        • Radziwon-Balicka A.
        • Jurasz P.
        • Michelakis E.
        • Jahroudi N.
        Hypoxia results in upregulation and de novo activation of von Willebrand factor expression in lung endothelial cells.
        Arterioscler. Thromb. Vasc. Biol. 2013; 33: 1329-1338
        • Aird W.C.
        • Edelberg J.M.
        • Weiler-Guettler H.
        • Simmons W.W.
        • Smith T.W.
        • Rosenberg R.D.
        Vascular bed-specific expression of an endothelial cell gene is programmed by the tissue microenvironment.
        J. Cell Biol. 1997; 138: 1117-1124
        • Aird W.C.
        • Jahroudi N.
        • Weiler-Guettler H.
        • Rayburn H.B.
        • Rosenberg R.D.
        Human von Willebrand factor gene sequences target expression to a subpopulation of endothelial cells in transgenic mice.
        Proc. Natl. Acad. Sci. U. S. A. 1995; 92: 4567-4571
        • Kleinschmidt A.M.
        • Nassiri M.
        • Stitt M.S.
        • Wasserloos K.
        • Watkins S.C.
        • Pitt B.R.
        • Jahroudi N.
        Sequences in intron 51 of the von Willebrand factor gene target promoter activation to a subset of lung endothelial cells in transgenic mice.
        J. Biol. Chem. 2008; 283: 2741-2750
        • Nassiri M.
        • Liu J.
        • Kulak S.
        • Uwiera R.R.
        • Aird W.C.
        • Ballermann B.J.
        • Jahroudi N.
        Repressors NFI and NFY participate in organ-specific regulation of von Willebrand factor promoter activity in transgenic mice.
        Arterioscler. Thromb. Vasc. Biol. 2010; 30: 1423-1429
        • Guan J.
        • Guillot P.V.
        • Aird W.C.
        Characterization of the mouse von Willebrand factor promoter.
        Blood. 1999; 94: 3405-3412
        • Liu J.
        • Yuan L.
        • Molema G.
        • Regan E.
        • Janes L.
        • Beeler D.
        • Spokes K.C.
        • Okada Y.
        • Minami T.
        • Oettgen P.
        • Aird W.C.
        Vascular bed-specific regulation of the von Willebrand factor promoter in the heart and skeletal muscle.
        Blood. 2011; 117: 342-351
        • Jahroudi N.
        • Schmaier A.
        • Srikanth S.
        • Mahdi F.
        • Lutka F.A.
        • Bowser R.
        Von Willebrand factor promoter targets the expression of amyloid beta protein precursor to brain vascular endothelial cells of transgenic mice.
        J. Alzheimers Dis. 2003; 5: 149-158
        • Jahroudi N.
        • Ardekani A.M.
        • Greenberger J.S.
        An NF1-like protein functions as a repressor of the von Willebrand factor promoter.
        J. Biol. Chem. 1996; 271: 21413-21421
        • Semenza G.L.
        • Agani F.
        • Iyer N.
        • Kotch L.
        • Laughner E.
        • Leung S.
        • Yu A.
        Regulation of cardiovascular development and physiology by hypoxia-inducible factor 1.
        Ann. N. Y. Acad. Sci. 1999; 874: 262-268
        • Jahroudi N.
        • Lynch D.C.
        Endothelial-cell-specific regulation of von Willebrand factor gene expression.
        Mol. Cell Biol. 1994; 14: 999-1008
        • Liu J.
        • Kanki Y.
        • Okada Y.
        • Jin E.
        • Yano K.
        • Shih S.C.
        • Minami T.
        • Aird W.C.
        A +220 GATA motif mediates basal but not endotoxin-repressible expression of the von Willebrand factor promoter in Hprt-targeted transgenic mice.
        J. Thromb. Haemostasis. 2009; 7: 1384-1392
        • Peng Y.
        • Jahroudi N.
        The NFY transcription factor inhibits von Willebrand factor promoter activation in non-endothelial cells through recruitment of histone deacetylases.
        J. Biol. Chem. 2003; 278: 8385-8394
        • Akman H.O.
        • Zhang H.
        • Siddiqui M.A.
        • Solomon W.
        • Smith E.L.
        • Batuman O.A.
        Response to hypoxia involves transforming growth factor-beta2 and Smad proteins in human endothelial cells.
        Blood. 2001; 98: 3324-3331
        • Froese N.
        • Kattih B.
        • Breitbart A.
        • Grund A.
        • Geffers R.
        • Molkentin J.D.
        • Kispert A.
        • Wollert K.C.
        • Drexler H.
        • Heineke J.
        GATA6 promotes angiogenic function and survival in endothelial cells by suppression of autocrine transforming growth factor beta/activin receptor-like kinase 5 signaling.
        J. Biol. Chem. 2011; 286: 5680-5690
        • Fish J.E.
        • Matouk C.C.
        • Yeboah E.
        • Bevan S.C.
        • Khan M.
        • Patil K.
        • Ohh M.
        • Marsden P.A.
        Hypoxia-inducible expression of a natural cis-antisense transcript inhibits endothelial nitric-oxide synthase.
        J. Biol. Chem. 2007; 282: 15652-15666
        • Tang J.L.
        • Zembowicz A.
        • Xu X.M.
        • Wu K.K.
        Role of Sp1 in transcriptional activation of human nitric oxide synthase type III gene.
        Biochem. Biophys. Res. Commun. 1995; 213: 673-680
        • Peng Y.
        • Stewart D.
        • Li W.
        • Hawkins M.
        • Kulak S.
        • Ballermann B.
        • Jahroudi N.
        Irradiation modulates association of NF-Y with histone-modifying cofactors PCAF and HDAC.
        Oncogene. 2007; 26: 7576-7583
        • Vaissiere T.
        • Sawan C.
        • Herceg Z.
        Epigenetic interplay between histone modifications and DNA methylation in gene silencing.
        Mutat. Res. 2008; 659: 40-48
        • Yuan L.
        • Chan G.C.
        • Beeler D.
        • Janes L.
        • Spokes K.C.
        • Dharaneeswaran H.
        • Mojiri A.
        • Adams W.J.
        • Sciuto T.
        • Garcia-Cardena G.
        • Molema G.
        • Kang P.M.
        • Jahroudi N.
        • Marsden P.A.
        • Dvorak A.
        • Regan E.R.
        • Aird W.C.
        A role of stochastic phenotype switching in generating mosaic endothelial cell heterogeneity.
        Nat. Commun. 2016; 7: 10160
        • Shirodkar A.V.
        • St Bernard R.
        • Gavryushova A.
        • Kop A.
        • Knight B.J.
        • Yan M.S.
        • Man H.S.
        • Sud M.
        • Hebbel R.P.
        • Oettgen P.
        • Aird W.C.
        • Marsden P.A.
        A mechanistic role for DNA methylation in endothelial cell (EC)-enriched gene expression: relationship with DNA replication timing.
        Blood. 2013; 121: 3531-3540
        • Kerkela R.
        • Karsikas S.
        • Szabo Z.
        • Serpi R.
        • Magga J.
        • Gao E.
        • Alitalo K.
        • Anisimov A.
        • Sormunen R.
        • Pietila I.
        • Vainio L.
        • Koch W.J.
        • Kivirikko K.I.
        • Myllyharju J.
        • Koivunen P.
        Activation of hypoxia response in endothelial cells contributes to ischemic cardioprotection.
        Mol. Cell Biol. 2013; 33: 3321-3329
        • Brill A.
        • Suidan G.L.
        • Wagner D.D.
        Hypoxia, such as encountered at high altitude, promotes deep vein thrombosis in mice.
        J. Thromb. Haemostasis. 2013; 11: 1773-1775
        • Chapin J.C.
        • Hajjar K.A.
        Fibrinolysis and the control of blood coagulation.
        Blood Rev. 2015; 29: 17-24
        • Hnisz D.
        • Abraham B.J.
        • Lee T.I.
        • Lau A.
        • Saint-Andre V.
        • Sigova A.A.
        • Hoke H.A.
        • Young R.A.
        Super-enhancers in the control of cell identity and disease.
        Cell. 2013; 155: 934-947
        • Singh B.
        • Biswas I.
        • Bhagat S.
        • Surya Kumari S.
        • Khan G.A.
        HMGB1 facilitates hypoxia-induced vWF upregulation through TLR2-MYD88-SP1 pathway.
        Eur. J. Immunol. 2016; 46: 2388-2400
        • Augustin H.G.
        • Koh G.Y.
        Organotypic vasculature: from descriptive heterogeneity to functional pathophysiology.
        Science. 2017; 357
        • Aird W.C.
        Phenotypic heterogeneity of the endothelium: II. Representative vascular beds.
        Circ. Res. 2007; 100: 174-190
        • Aird W.C.
        Phenotypic heterogeneity of the endothelium: I. Structure, function, and mechanisms.
        Circ. Res. 2007; 100: 158-173
        • Nilsson I.
        • Shibuya M.
        • Wennstrom S.
        Differential activation of vascular genes by hypoxia in primary endothelial cells.
        Exp. Cell Res. 2004; 299: 476-485
        • Dmitrieva N.I.
        • Burg M.B.
        Secretion of von Willebrand factor by endothelial cells links sodium to hypercoagulability and thrombosis.
        Proc. Natl. Acad. Sci. U. S. A. 2014; 111: 6485-6490
        • Farrell A.J.
        • Williams R.B.
        • Stevens C.R.
        • Lawrie A.S.
        • Cox N.L.
        • Blake D.R.
        Exercise induced release of von Willebrand factor: evidence for hypoxic reperfusion microvascular injury in rheumatoid arthritis.
        Ann. Rheum. Dis. 1992; 51: 1117-1122
        • Kiouptsi K.
        • Gambaryan S.
        • Walter E.
        • Walter U.
        • Jurk K.
        • Reinhardt C.
        Hypoxia impairs agonist-induced integrin alphaIIbbeta3 activation and platelet aggregation.
        Sci. Rep. 2017; 7: 7621
        • Pinsky D.J.
        • Naka Y.
        • Liao H.
        • Oz M.C.
        • Wagner D.D.
        • Mayadas T.N.
        • Johnson R.C.
        • Hynes R.O.
        • Heath M.
        • Lawson C.A.
        • Stern D.M.
        Hypoxia-induced exocytosis of endothelial cell Weibel-Palade bodies. A mechanism for rapid neutrophil recruitment after cardiac preservation.
        J. Clin. Invest. 1996; 97: 493-500
        • Suidan G.L.
        • Brill A.
        • De Meyer S.F.
        • Voorhees J.R.
        • Cifuni S.M.
        • Cabral J.E.
        • Wagner D.D.
        Endothelial Von Willebrand factor promotes blood-brain barrier flexibility and provides protection from hypoxia and seizures in mice.
        Arterioscler. Thromb. Vasc. Biol. 2013; 33: 2112-2120
        • Rocke A.S.
        • Paterson G.G.
        • Barber M.T.
        • Jackson A.I.R.
        • Main S.
        • Stannett C.
        • Schnopp M.F.
        • Baillie J.K.
        • Horne E.H.
        • Moores C.
        • Harrison P.
        • Nimmo A.F.
        • Thompson A.A.R.
        Thromboelastometry and platelet function during acclimatization to high altitude.
        Thromb. Haemostasis. 2018; 118: 63-71
        • Mackman N.
        New insights into the mechanisms of venous thrombosis.
        J. Clin. Invest. 2012; 122: 2331-2336
        • Ogawa S.
        • Shreeniwas R.
        • Brett J.
        • Clauss M.
        • Furie M.
        • Stern D.M.
        The effect of hypoxia on capillary endothelial-cell function - modulation of barrier and coagulant function.
        Br. J. Haematol. 1990; 75: 517-524
        • Minami T.
        • Donovan D.J.
        • Tsai J.C.
        • Rosenberg R.D.
        • Aird W.C.
        Differential regulation of the von Willebrand factor and Flt-1 promoters in the endothelium of hypoxanthine phosphoribosyltransferase-targeted mice.
        Blood. 2002; 100: 4019-4025