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Thromb Haemost 2014; 112(02): 390-401
DOI: 10.1160/TH13-09-0773
Cardiovascular Biology and Cell Signalling
Schattauer GmbH

Titration of the gap junction protein Connexin43 reduces atherogenesis

Sandrine Morel
1   Department of Pathology and Immunology, University of Geneva, Geneva, Switzerland
2   Department of Internal Medicine – Division of Cardiology, University of Geneva, Geneva, Switzerland
,
Marc Chanson
3   Laboratory of Clinical Investigation III, Department of Pediatrics, Geneva University Hospitals and University of Geneva, Geneva, Switzerland
,
Thien D. Nguyen
4   Department of Microbiology, SUNY Upstate Medical University, Syracuse, New York, USA
,
Aaron M. Glass
4   Department of Microbiology, SUNY Upstate Medical University, Syracuse, New York, USA
,
Maya Z. Richani Sarieddine
3   Laboratory of Clinical Investigation III, Department of Pediatrics, Geneva University Hospitals and University of Geneva, Geneva, Switzerland
,
Merlijn J. Meens
1   Department of Pathology and Immunology, University of Geneva, Geneva, Switzerland
2   Department of Internal Medicine – Division of Cardiology, University of Geneva, Geneva, Switzerland
,
Laurent Burnier
1   Department of Pathology and Immunology, University of Geneva, Geneva, Switzerland
2   Department of Internal Medicine – Division of Cardiology, University of Geneva, Geneva, Switzerland
,
Brenda R. Kwak
1   Department of Pathology and Immunology, University of Geneva, Geneva, Switzerland
2   Department of Internal Medicine – Division of Cardiology, University of Geneva, Geneva, Switzerland
,
Steven M. Taffet
4   Department of Microbiology, SUNY Upstate Medical University, Syracuse, New York, USA
› Author Affiliations Financial support: This work was supported by grants from the Swiss National Science Foundation (310030–127551 and 310030–143343 to B.R. Kwak and 310030–119739 to M. Chanson), the Fondation Novartis (to B.R. Kwak) and the National Institutes of Health (#HL100111 to S.M. Taffet).
Further Information

Publication History

Received: 19 September 2013

Accepted after major revision: 21 March 2014

Publication Date:
21 November 2017 (online)

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Summary

Ubiquitous reduction of the gap junction protein Connexin43 (Cx43) in mice provides beneficial effects on progression and composition of atherosclerotic lesions. Cx43 is expressed in multiple atheroma-associated cells but its function in each cell type is not known. To examine specifically the role of Cx43 in immune cells, we have lethally irradiated low-density lipoprotein receptor-deficient mice and reconstituted with Cx43+/+, Cx43+/− or Cx43−/− haematopoietic fetal liver cells. Progression of atherosclerosis was significantly lower in aortic roots of Cx43+/− chimeras compared with Cx43+/+ and Cx43−/− chimeras, and their plaques contained significantly less neutrophils. The relative proportion of circulating leukocytes was similar between the three groups. Interestingly, the chemoattraction of neutrophils, which did not express Cx43, was reduced in response to supernatant secreted by Cx43+/− macrophages in comparison with the ones of Cx43+/+ and Cx43−/− macrophages. Cx43+/− macrophages did not differ from Cx43+/+ and Cx43−/− macrophages in terms of M1/M2 polarisation but show modified gene expression for a variety chemokines and complement components. In conclusion, titration of Cx43 expression in bone marrow-derived macrophages reduces atherosclerotic plaque formation and chemoattraction of neutrophils to the lesions.

Keywords

Atherosclerosis - Connexin43 - neutrophils - macrophages
 

  • References

  • 1 Libby P. Mechanisms of acute coronary syndromes and their implications for therapy. N Engl J Med 2013; 368: 2004-2013.
    CrossrefPubMedSearch in Google Scholar
  • 2 Weber C, Noels H. Atherosclerosis: current pathogenesis and therapeutic options. Nat Med 2011; 17: 1410-1422.
    CrossrefPubMedSearch in Google Scholar
  • 3 Hansson GK, Hermansson A. The immune system in atherosclerosis. Nat Immunol 2011; 12: 204-212.
    CrossrefPubMedSearch in Google Scholar
  • 4 Lahoute C, Herbin O, Mallat Z. et al. Adaptive immunity in atherosclerosis: mechanisms and future therapeutic targets. Nat Rev Cardiol 2011; 08: 348-358.
    CrossrefPubMedSearch in Google Scholar
  • 5 van Leeuwen M, Gijbels MJ, Duijvestijn A. et al. Accumulation of myeloperoxidase-positive neutrophils in atherosclerotic lesions in LDLR−/− mice. Arterioscler Thromb Vasc Biol 2008; 28: 84-89.
    CrossrefPubMedSearch in Google Scholar
  • 6 Drechsler M, Megens RT, van Zandvoort M. et al. Hyperlipidemia-triggered neutrophilia promotes early atherosclerosis. Circulation 2010; 122: 1837-1845.
    CrossrefPubMedSearch in Google Scholar
  • 7 Zernecke A, Bot I, Djalali-Talab Y. et al. Protective role of CXC receptor 4/CXC ligand 12 unveils the importance of neutrophils in atherosclerosis. Circ Res 2008; 102: 209-217.
    CrossrefPubMedSearch in Google Scholar
  • 8 Rotzius P, Thams S, Soehnlein O. et al. Distinct infiltration of neutrophils in lesion shoulders in ApoE−/− mice. Am J Pathol 2010; 177: 493-500.
    CrossrefPubMedSearch in Google Scholar
  • 9 Zernecke A, Weber C. Chemokines in Atherosclerosis: Proceedings Resumed. Arterioscler Thromb Vasc Biol. 2014 Epub ahead of print
    PubMedSearch in Google Scholar
  • 10 de Jager SC, Bot I, Kraaijeveld AO. et al. Leukocyte-specific CCL3 deficiency inhibits atherosclerotic lesion development by affecting neutrophil accumulation. Arterioscler Thromb Vasc Biol 2013; 33: e75-83.
    CrossrefPubMedSearch in Google Scholar
  • 11 Lenglet S, Thomas A, Soehnlein O. et al. Fatty acid amide hydrolase deficiency enhances intraplaque neutrophil recruitment in atherosclerotic mice. Arterioscler Thromb Vasc Biol 2013; 33: 215-223.
    CrossrefPubMedSearch in Google Scholar
  • 12 Doring Y, Soehnlein O, Drechsler M. et al. Haematopoietic interferon regulatory factor 8-deficiency accelerates atherosclerosis in mice. Arterioscler Thromb Vasc Biol 2012; 32: 1613-1623.
    CrossrefPubMedSearch in Google Scholar
  • 13 Pfenniger A, Chanson M, Kwak BR. Connexins in atherosclerosis. Biochim Biophys Acta 2013; 1828: 157-166.
    CrossrefPubMedSearch in Google Scholar
  • 14 Chadjichristos CE, Scheckenbach KE, van Veen TA. et al. Endothelial-specific deletion of connexin40 promotes atherosclerosis by increasing CD73-depend-ent leukocyte adhesion. Circulation 2010; 121: 123-131.
    CrossrefPubMedSearch in Google Scholar
  • 15 Wong CW, Christen T, Roth I. et al. Connexin37 protects against atherosclerosis by regulating monocyte adhesion. Nat Med 2006; 12: 950-954.
    CrossrefPubMedSearch in Google Scholar
  • 16 Kwak BR, Veillard N, Pelli G. et al. Reduced connexin43 expression inhibits atherosclerotic lesion formation in low-density lipoprotein receptor-deficient mice. Circulation 2003; 107: 1033-1039.
    CrossrefPubMedSearch in Google Scholar
  • 17 Blackburn JP, Peters NS, Yeh HI. et al. Up-regulation of connexin43 gap junctions during early stages of human coronary atherosclerosis. Arterioscler Thromb Vasc Biol 1995; 15: 1219-1228.
    CrossrefPubMedSearch in Google Scholar
  • 18 Polacek D, Bech F, McKinsey JF. et al. Connexin43 gene expression in the rabbit arterial wall: effects of hypercholesterolemia, balloon injury and their combination. J Vasc Res 1997; 34: 19-30.
    CrossrefPubMedSearch in Google Scholar
  • 19 Kwak BR, Mulhaupt F, Veillard N. et al. Altered pattern of vascular connexin expression in atherosclerotic plaques. Arterioscler Thromb Vasc Biol 2002; 22: 225-230.
    CrossrefPubMedSearch in Google Scholar
  • 20 Ishibashi S, Goldstein JL, Brown MS. et al. Massive xanthomatosis and atherosclerosis in cholesterol-fed low density lipoprotein receptor-negative mice. J Clin Invest 1994; 93: 1885-1893.
    CrossrefPubMedSearch in Google Scholar
  • 21 Nguyen TD, Taffet SM. A model system to study Connexin 43 in the immune system. Mol Immunol 2009; 46: 2938-2946.
    CrossrefPubMedSearch in Google Scholar
  • 22 Glass AM, Wolf BJ, Schneider KM. et al. Connexin43 is dispensable for phagocytosis. J Immunol 2013; 190: 4830-4835.
    CrossrefPubMedSearch in Google Scholar
  • 23 Sarieddine MZ, Scheckenbach KE, Foglia B. et al. Connexin43 modulates neutrophil recruitment to the lung. J Cell Mol Med 2009; 13: 4560-4570.
    CrossrefPubMedSearch in Google Scholar
  • 24 Gonzalez-Nieto D, Li L, Kohler A. et al. Connexin-43 in the osteogenic BM niche regulates its cellular composition and the bidirectional traffic of haematopoietic stem cells and progenitors. Blood 2012; 119: 5144-5154.
    CrossrefPubMedSearch in Google Scholar
  • 25 Moore KJ, Tabas I. Macrophages in the pathogenesis of atherosclerosis. Cell 2011; 145: 341-355.
    CrossrefPubMedSearch in Google Scholar
  • 26 Zahler S, Hoffmann A, Gloe T. et al. Gap-junctional coupling between neutrophils and endothelial cells: a novel modulator of transendothelial migration. J Leukoc Biol 2003; 73: 118-126.
    CrossrefPubMedSearch in Google Scholar
  • 27 Branes MC, Contreras JE, Saez JC. Activation of human polymorphonuclear cells induces formation of functional gap junctions and expression of connexins. Med Sci Monit 2002; 08: BR313-323.
    PubMedSearch in Google Scholar
  • 28 Eltzschig HK, Eckle T, Mager A. et al. ATP release from activated neutrophils occurs via connexin 43 and modulates adenosine-dependent endothelial cell function. Circ Res 2006; 99: 1100-1108.
    CrossrefPubMedSearch in Google Scholar
  • 29 Eltzschig HK, Thompson LF, Karhausen J. et al. Endogenous adenosine produced during hypoxia attenuates neutrophil accumulation: coordination by extracellular nucleotide metabolism. Blood 2004; 104: 3986-3992.
    CrossrefPubMedSearch in Google Scholar
  • 30 Scerri I, Tabary O, Dudez T. et al. Gap junctional communication does not contribute to the interaction between neutrophils and airway epithelial cells. Cell Commun Adhes 2006; 13: 1-12.
    CrossrefPubMedSearch in Google Scholar
  • 31 Cherian PP, Siller-Jackson AJ, Gu S. et al. Mechanical strain opens connexin 43 hemichannels in osteocytes: a novel mechanism for the release of prostaglandin. Mol Biol Cell 2005; 16: 3100-3106.
    CrossrefPubMedSearch in Google Scholar
  • 32 Burra S, Nicolella DP, Francis WL. et al. Dendritic processes of osteocytes are mechanotransducers that induce the opening of hemichannels. Proc Natl Acad Sci USA 2010; 107: 13648-13653.
    CrossrefPubMedSearch in Google Scholar
  • 33 Batra N, Burra S, Siller-Jackson AJ. et al. Mechanical stress-activated integrin alpha5beta1 induces opening of connexin 43 hemichannels. Proc Natl Acad Sci USA 2012; 109: 3359-3364.
    CrossrefPubMedSearch in Google Scholar
  • 34 Wynn TA, Chawla A, Pollard JW. Macrophage biology in development, homeostasis and disease. Nature 2013; 496: 445-455.
    CrossrefPubMedSearch in Google Scholar
  • 35 Kameritsch P, Pogoda K, Pohl U. Channel-independent influence of connexin 43 on cell migration. Biochim Biophys Acta 2012; 1818: 1993-2001.
    CrossrefPubMedSearch in Google Scholar
  • 36 Reaume AG, de Sousa PA, Kulkarni S. et al. Cardiac malformation in neonatal mice lacking connexin43. Science 1995; 267: 1831-1834.
    CrossrefPubMedSearch in Google Scholar
  • 37 Lo CW, Waldo KL, Kirby ML. Gap junction communication and the modulation of cardiac neural crest cells. Trends Cardiovasc Med 1999; 09: 63-69.
    CrossrefPubMedSearch in Google Scholar
  • 38 Sullivan R, Huang GY, Meyer RA. et al. Heart malformations in transgenic mice exhibiting dominant negative inhibition of gap junctional communication in neural crest cells. Dev Biol 1998; 204: 224-234.
    CrossrefPubMedSearch in Google Scholar
  • 39 Huang GY, Cooper ES, Waldo K. et al. Gap junction-mediated cell-cell communication modulates mouse neural crest migration. J Cell Biol 1998; 143: 1725-1734.
    CrossrefPubMedSearch in Google Scholar
  • 40 Chadjichristos CE, Matter CM, Roth I. et al. Reduced connexin43 expression limits neointima formation after balloon distension injury in hypercholesterolemic mice. Circulation 2006; 113: 2835-2843.
    CrossrefPubMedSearch in Google Scholar
  • 41 Chadjichristos CE, Morel S, Derouette JP. et al. Targeting connexin 43 prevents platelet-derived growth factor-BB-induced phenotypic change in porcine coronary artery smooth muscle cells. Circ Res 2008; 102: 653-660.
    CrossrefPubMedSearch in Google Scholar
  • 42 Liao Y, Regan CP, Manabe I. et al. Smooth muscle-targeted knockout of connexin43 enhances neointimal formation in response to vascular injury. Arterioscler Thromb Vasc Biol 2007; 27: 1037-1042.
    CrossrefPubMedSearch in Google Scholar
  • 43 Theis M, de Wit C, Schlaeger TM. et al. Endothelium-specific replacement of the connexin43 coding region by a lacZ reporter gene. Genesis 2001; 29: 1-13.
    CrossrefPubMedSearch in Google Scholar