Download PDF
Thromb Haemost 2012; 108(04): 742-749
DOI: 10.1160/TH12-05-0285
Cardiovascular Biology and Cell Signalling
Schattauer GmbH

Atrial fibrillation in pigs induces left atrial endocardial transcriptional remodelling

Jorge Cerveró
1   Division of Cardiovascular Sciences, Laboratory of Thrombosis and Haemostasis, Centre for Applied Medical Research, University of Navarra, Pamplona, Spain
,
Víctor Segura
2   Proteomics, Genomics & Bioinformatics Unit, Centre for Applied Medical Research, University of Navarra, Pamplona, Spain
,
Alfonso Macías
3   Department of Cardiology and Cardiovascular Surgery, Clínica Universidad de Navarra, Pamplona, Spain
,
Juan José Gavira
3   Department of Cardiology and Cardiovascular Surgery, Clínica Universidad de Navarra, Pamplona, Spain
,
Ramón Montes
1   Division of Cardiovascular Sciences, Laboratory of Thrombosis and Haemostasis, Centre for Applied Medical Research, University of Navarra, Pamplona, Spain
,
José Hermida
1   Division of Cardiovascular Sciences, Laboratory of Thrombosis and Haemostasis, Centre for Applied Medical Research, University of Navarra, Pamplona, Spain
› Author Affiliations Financial support: This work was supported through the Unión Temporal de Empresas project CIMA and by grants from Instituto de Salud Carlos III (PI08/1349, PI11/01458 and Red Temática de Investigación RECAVA RD06/0014/0008) and Health Department, Gobierno de Navarra (15/09). Jorge Cerveró was supported by a grant from the Sociedad Española de Trombosis y Hemostasia.
Further Information

Publication History

Received: 03 May 2012

Accepted after major revision: 21 July 2012

Publication Date:
29 November 2017 (online)

    Permissions and Reprints

Summary

The leading cause of cardioembolic stroke is atrial fibrillation (AF), which predisposes to atrial thrombus formation. Although rheological alterations promote a hypercoagulable environment, as yet undefined factors contribute to thrombogenesis. The role of the endocardium has barely been explored. To approach this topic, rapid atrial pacing (RAP) was applied in four pigs to mimic A F. Left and right endocardial cells were isolated separately and their gene expression pattern was compared with that of four control pigs. The AF-characteristic rhythm disorders and endothelial nitric oxide synthase down-regulation were successfully reproduced, and validated RAP to mimic A F. A change was observed in the transcriptomic endocardial profile after RAP: the expression of 364 genes was significantly altered (p<0.01), 29 of them having passed the B>0 criteria. The left atrial endocardium [325 genes (7 genes, B>0)] was largely responsible for such alterations. Blood coagulation, blood vessel morphogenesis and inflammatory response are among the most significant altered functions, and help to explain the activation of coagulation observed after RAP: D-dimer, 0.49 (1.63) vs. 0.23 (0.24) mg/l [median (interquartile range)] in controls, p=0.02. Furthermore, three genes directly related to thrombotic processes were differentially expressed after RAP: FGL2 [fold change (FC)=0.85; p=0.007], APLP2 (FC=-0.47; p=0.005) and ADAMTS-18 (FC=-0.69; p=0.004). We demonstrate for the first time that AF induces a global expression change in the left atrial endocardium associated with an activation of blood coagulation. The nature of some of the altered functions and genes provides clues to identify new therapeutic targets.

Keywords

Animals models - endothelial cells - vascular remodelling - heart - gene expression

 

  • References

  • 1 Hart RG, Halperin JL. Atrial fibrillation and stroke: Concepts and controversies. Stroke 2001; 32: 803-808.
    CrossrefPubMedGoogle Scholar
  • 2 Wolf PA, Abbott RD, Kannel WB. Atrial fibrillation as an independent risk factor for stroke: The framingham study. Stroke 1991; 22: 983-988.
    CrossrefPubMedGoogle Scholar
  • 3 Steinberg JS. Atrial fibrillation: An emerging epidemic?. Heart 2004; 90: 239-240.
    CrossrefPubMedGoogle Scholar
  • 4 Miyasaka Y, Barnes ME, Gersh BJ. et al. Secular trends in incidence of atrial fibrillation in olmsted county, minnesota, 1980 to 2000, and implications on the projections for future prevalence. Circulation 2006; 114: 119-125.
    CrossrefPubMedGoogle Scholar
  • 5 Freestone B, Lip GY. The endothelium and atrial fibrillation the prothrombotic state revisited. Hamostaseologie 2008; 28: 207-212.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 6 Feng D, D'Agostino RB, Silbershatz H. et al. Hemostatic state and atrial fibrillation (the framingham offspring study). Am J Cardiol 2001; 87: 168-171.
    CrossrefPubMedGoogle Scholar
  • 7 Mitusch R, Siemens HJ, Garbe M. et al. Detection of a hypercoagulable state in nonvalvular atrial fibrillation and the effect of anticoagulant therapy. Thromb Haemost 1996; 75: 219-223.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 8 Aird WC. Phenotypic heterogeneity of the endothelium: II representative vascular beds. Circ Res 2007; 100: 174-190.
    CrossrefPubMedGoogle Scholar
  • 9 Ryan US, Mortara M, Whitaker C. Methods for microcarrier culture of bovine pulmonary artery endothelial cells avoiding the use of enzymes. Tissue Cell 1980; 12: 619-635.
    CrossrefPubMedGoogle Scholar
  • 10 Irizarry RA, Bolstad BM, Collin F. et al. Summaries of affymetrix GeneChip probe level data. Nucleic Acids Res 2003; 31: e15.
    CrossrefPubMedGoogle Scholar
  • 11 Smyth GK. Linear models and empirical bayes methods for assessing differential expression in microarray experiments. Stat Appl Genet Mol Biol. 2004 03. Article 3.
    PubMedGoogle Scholar
  • 12 Gentleman R, Carey V, Huber W. et al. Bioinformatics and computational biology solutions using R and bioconductor. New York: Springer; 2005
    Google Scholar
  • 13 Draghici S. Data analysis tools for DNA microarrays. London: Chapman Hall/ CRC; 2003
    Google Scholar
  • 14 Cai H, Li Z, Goette A. et al. Downregulation of endocardial nitric oxide synthase expression and nitric oxide production in atrial fibrillation: Potential mechanisms for atrial thrombosis and stroke. Circulation 2002; 106: 2854-2858.
    CrossrefPubMedGoogle Scholar
  • 15 Akar JG, Jeske W, Wilber DJ. Acute onset human atrial fibrillation is associated with local cardiac platelet activation and endothelial dysfunction. J Am Coll Cardiol 2008; 51: 1790-1793.
    CrossrefPubMedGoogle Scholar
  • 16 Sadanaga T, Sadanaga M, Ogawa S. Evidence that D-dimer levels predict subsequent thromboembolic and cardiovascular events in patients with atrial fibrillation during oral anticoagulant therapy. J Am Coll Cardiol 2010; 55: 2225-2231.
    CrossrefPubMedGoogle Scholar
  • 17 Wang J, Klysik E, Sood S. et al. Pitx2 prevents susceptibility to atrial arrhythmias by inhibiting left-sided pacemaker specification. Proc Natl Acad Sci USA 2010; 107: 9753-9758.
    CrossrefPubMedGoogle Scholar
  • 18 Lin CS, Lai L P, Lin JL. et al. Increased expression of extracellular matrix proteins in rapid atrial pacing-induced atrial fibrillation. Heart Rhythm 2007; 04: 938-949.
    CrossrefPubMedGoogle Scholar
  • 19 Lai L P, Lin JL, Lin CS. et al. Functional genomic study on atrial fibrillation using cDNA microarray and two-dimensional protein electrophoresis techniques and identification of the myosin regulatory light chain isoform reprogramming in atrial fibrillation. J Cardiovasc Electrophysiol 2004; 15: 214-223.
    CrossrefPubMedGoogle Scholar
  • 20 Kim NH, Ahn Y, Oh SK. et al. Altered patterns of gene expression in response to chronic atrial fibrillation. Int Heart J 2005; 46: 383-395.
    CrossrefPubMedGoogle Scholar
  • 21 Hua G, Zhang Q, Fan Z. Heat shock protein 75 (TRAP1) antagonizes reactive oxygen species generation and protects cells from granzyme M-mediated apoptosis. J Biol Chem 2007; 282: 20553-20560.
    CrossrefPubMedGoogle Scholar
  • 22 Mattiussi S, Turrini P, Testolin L. et al. p21(Waf1/Cip1/Sdi1) mediates shear stress-dependent antiapoptotic function. Cardiovasc Res 2004; 61: 693-704.
    CrossrefPubMedGoogle Scholar
  • 23 Conway DS, Buggins P, Hughes E. et al. Prognostic significance of raised plasma levels of interleukin-6 and C-reactive protein in atrial fibrillation. Am Heart J 2004; 148: 462-466.
    CrossrefPubMedGoogle Scholar
  • 24 Hermida J, Lopez FL, Montes R. et al. Usefulness of high-sensitivity C-reactive protein to predict mortality in patients with atrial fibrillation (from the atherosclerosis risk in communities [ARIC] study). Am J Cardiol 2012; 109: 95-99.
    CrossrefPubMedGoogle Scholar
  • 25 Patel P, Dokainish H, Tsai P. et al. Update on the association of inflammation and atrial fibrillation. J Cardiovasc Electrophysiol 2010; 21: 1064-1070.
    CrossrefPubMedGoogle Scholar
  • 26 Caposio P, Gugliesi F, Zannetti C. et al. A novel role of the interferon-inducible protein IFI16 as inducer of proinflammatory molecules in endothelial cells. J Biol Chem 2007; 282: 33515-33529.
    CrossrefPubMedGoogle Scholar
  • 27 Okada M, Suzuki K, Takada K. et al. Detection o up-regulated genes in thrombin stimulated human umbilical vein endothelial cells. Thromb Res 2006; 118: 715-721.
    CrossrefPubMedGoogle Scholar
  • 28 Pullamsetti SS, Savai R, Schaefer MB. et al. cAMP phosphodiesterase inhibitors increases nitric oxide production by modulating dimethylarginine dimethylami-nohydrolases. Circulation 2011; 123: 1194-1204.
    CrossrefPubMedGoogle Scholar
  • 29 Gretarsdottir S, Thorleifsson G, Reynisdottir ST. et al. The gene encoding phosphodiesterase 4D confers risk of ischemic stroke. Nat Genet 2003; 35: 131-138.
    CrossrefPubMedGoogle Scholar
  • 30 Pan H, Richards AA, Zhu X. et al. A novel mutation in LAMIN A/C is associated with isolated early-onset atrial fibrillation and progressive atrioventricular block followed by cardiomyopathy and sudden cardiac death. Heart Rhythm 2009; 06: 707-710.
    CrossrefPubMedGoogle Scholar
  • 31 Davis ME, Cai H, Drummond GR. et al. Shear stress regulates endothelial nitric oxide synthase expression through c-src by divergent signaling pathways. Circ Res 2001; 89: 1073-1080.
    CrossrefPubMedGoogle Scholar
  • 32 Gu M, Brecher P. Nitric oxide-induced increase in p21(Sdi1/Cip1/Waf1) expression during the cell cycle in aortic adventitial fibroblasts. Arterioscler Thromb Vasc Biol 2000; 20: 27-34.
    CrossrefPubMedGoogle Scholar
  • 33 Cervero J, Montes R, Espana F. et al. Limited ability to activate protein C confers left atrial endocardium a thrombogenic phenotype: A role in cardioembolic stroke?. Stroke 2011; 42: 2622-2624.
    CrossrefPubMedGoogle Scholar
  • 34 Ghanekar A, Mendicino M, Liu H. et al. Endothelial induction of fgl2 contributes to thrombosis during acute vascular xenograft rejection. J Immunol 2004; 172: 5693-5701.
    CrossrefPubMedGoogle Scholar
  • 35 Xu F, Previti ML, Nieman MT. et al. AbetaPP/APLP2 family of kunitz serine proteinase inhibitors regulate cerebral thrombosis. J Neurosci 2009; 29: 5666-5670.
    CrossrefPubMedGoogle Scholar
  • 36 Li Z, Nardi MA, Li YS. et al. C-terminal ADAMTS-18 fragment induces oxidative platelet fragmentation, dissolves platelet aggregates, and protects against carotid artery occlusion and cerebral stroke. Blood 2009; 113: 6051-6060.
    CrossrefPubMedGoogle Scholar
  • 37 Goette A, Bukowska A, Lendeckel U. et al. Angiotensin II receptor blockade reduces tachycardia-induced atrial adhesion molecule expression. Circulation 2008; 117: 732-742.
    CrossrefPubMedGoogle Scholar