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Thromb Haemost 2015; 113(03): 655-663
DOI: 10.1160/TH14-05-0466
Atherosclerosis and Ischaemic Disease
Schattauer GmbH

An integrated genomic-transcriptomic approach supports a role for the proto-oncogene BCL3 in atherosclerosis

Giovanna Marchetti
1   Department of Biomedical and Specialty Surgical Sciences, University of Ferrara, Ferrara, Italy
,
Domenico Girelli
2   Department of Medicine, University of Verona, Verona, Italy
,
Carlotta Zerbinati
1   Department of Biomedical and Specialty Surgical Sciences, University of Ferrara, Ferrara, Italy
,
Barbara Lunghi
3   Department of Life Sciences and Biotechnology, University of Ferrara, Ferrara, Italy
,
Simonetta Friso
2   Department of Medicine, University of Verona, Verona, Italy
,
Silvia Meneghetti
1   Department of Biomedical and Specialty Surgical Sciences, University of Ferrara, Ferrara, Italy
,
Matteo Coen
4   Department of Pathology and Immunology, University of Geneva, Geneva, Switzerland
,
Teresa Gagliano
5   Department of Medical Sciences, University of Ferrara, Ferrara, Italy
,
Giuseppe Guastella
1   Department of Biomedical and Specialty Surgical Sciences, University of Ferrara, Ferrara, Italy
,
Marie-Luce Bochaton-Piallat
4   Department of Pathology and Immunology, University of Geneva, Geneva, Switzerland
,
Francesca Pizzolo
2   Department of Medicine, University of Verona, Verona, Italy
,
Francesco Mascoli
6   Unit of Vascular and Endovascular Surgery, S. Anna University-Hospital, Ferrara, Italy
,
Giovanni Malerba
7   Department of Life and Reproduction Sciences, Section of Biology and Genetics, University of Verona, Verona, Italy
,
Matteo Bovolenta
3   Department of Life Sciences and Biotechnology, University of Ferrara, Ferrara, Italy
,
Manuela Ferracin
8   Department of Morphology, Surgery and Experimental Medicine, University of Ferrara, Ferrara, Italy
9   Laboratory for Technologies of Advanced Therapies (LTTA), University of Ferrara, Ferrara, Italy
,
Oliviero Olivieri
2   Department of Medicine, University of Verona, Verona, Italy
,
Francesco Bernardi
3   Department of Life Sciences and Biotechnology, University of Ferrara, Ferrara, Italy
,
Nicola Martinelli
2   Department of Medicine, University of Verona, Verona, Italy
› Author Affiliations Financial support: This study was supported by grants from the Italian Ministry of University and Research; the Veneto Region; the Cariverona Foundation, Verona; the Carife Foundation, Ferrara; the Italian Ministry of Health (Finalized Research “Emilia Romagna Region”).
Further Information

Publication History

Received: 26 May 2014

Accepted after major revision: 08 October 2014

Publication Date:
29 November 2017 (online)

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Summary

association studies of coronary artery disease (CAD), could include functionally relevant associations. We propose an integrated genomic and transcriptomic approach for unravelling new potential genetic signatures of atherosclerosis. Fifteen among 91 single nucleotide polymorphisms (SNPs) were first selected for association in a sex- and age-adjusted model by examining 510 patients with CAD and myocardial infarction and 388 subjects with normal coronary arteries (CAD-free) in the replication stages of a genome-wide association study. We investigated the expression of 71 genes proximal to the 15 tag-SNPs by two subsequent steps of microarray-based Mrna profiling, the former in vascular smooth muscle cell populations, isolated from non-atherosclerotic and atherosclerotic human carotid portions, and the latter in whole carotid specimens. BCL3 and PVRL2, contiguously located on chromosome 19, and ABCA1, extensively investigated before, were found to be differentially expressed. BCL3 and PVRL2 SNPs were genotyped within a second population of CAD patients (n=442) and compared with CAD-free subjects (n=393). The carriership of the BCL3 rs2965169 G allele was more represented among CAD patients and remained independently associated with CAD after adjustment for all the traditional cardiovascular risk factors (odds ratio=1.70 with 95% confidence interval 1.07–2.71), while the BCL3 rs8100239 A allele correlated with metabolic abnormalities. The upregulation of BCL3 mRNA levels in atherosclerotic tissue samples was consistent with BCL3 protein expression, which was detected by immunostaining in the intima-media of atherosclerotic specimens, but not within non-atherosclerotic ones. Our integrated approach suggests a role for BCL3 in cardiovascular diseases.

Keywords

BCL3 - genomics - transcriptomics - coronary artery disease - metabolic disorders

 

  • References

  • 1 Kathiresan S, Srivastava D. Genetics of human cardiovascular disease. Cell 2012; 148: 1242-1257.
    CrossrefPubMedGoogle Scholar
  • 2 Roger VL. et al. American Heart Association Statistics Committee and Stroke Statistics Subcommittee. Executive summary: heart disease and stroke statistics-2012 update: a report from the American Heart Association. Circulation 2012; 125: 188-197.
    CrossrefPubMedGoogle Scholar
  • 3 Ding K, Kullo IJ. Genome-wide association studies for atherosclerotic vascular disease and its risk factors. Circ Cardiovasc Genet 2009; 2: 63-72.
    CrossrefPubMedGoogle Scholar
  • 4 O’Donnell CJ, Nabel EG. Genomics of cardiovascular disease. N Engl J Med 2011; 365: 2098-2109.
    CrossrefPubMedGoogle Scholar
  • 5 Bijnens APJJ. et al. Genome-wide expression studies of atherosclerosis-Critical issue in methodology, analysis, interpretation of transcriptomics data. Arte-rioscler Thromb Vasc Biol 2006; 26: 1226-1235.
    CrossrefPubMedGoogle Scholar
  • 6 Miller DT. et al. Atherosclerosis: the path from genomics to therapeutics. J Am Coll Cardiol 2007; 49: 1589-1599.
    CrossrefPubMedGoogle Scholar
  • 7 Cappola TP, Margulies KB. Functional genomics applied to cardiovascular medicine. Circulation 2011; 124: 87-94.
    CrossrefPubMedGoogle Scholar
  • 8 Park SW. et al. Post-transcriptional regulation of low density lipoprotein receptor protein by proprotein convertase subtilisin/kexin type 9a in mouse liver. J Biol Chem 2004; 279: 50630-50638.
    CrossrefPubMedGoogle Scholar
  • 9 Lambert G. et al. Molecular basis of PCSK9 function. Atherosclerosis 2009; 203: 1-7.
    CrossrefPubMedGoogle Scholar
  • 10 Musunuru K. et al. From noncoding variant to phenotype via SORT1 at the 1p13 cholesterol locus. Nature 2010; 466: 714-719.
    CrossrefPubMedGoogle Scholar
  • 11 Yang X. Use of functional genomics to identify candidate genes underlying human genetic association studies of vascular diseases. Arterioscler Thromb Vasc Biol 2012; 32: 216-222.
    CrossrefPubMedGoogle Scholar
  • 12 Marian AJ, Belmont J. Strategic approaches to unravelling genetic causes of cardiovascular diseases. Circ Res 2011; 108: 1252-1269.
    CrossrefPubMedGoogle Scholar
  • 13 Stahl EA. et al. Bayesian inference analyses of the polygenic architecture of rheumatoid arthritis. Nat Genet 2012; 44: 483-489.
    CrossrefPubMedGoogle Scholar
  • 14 Park JH. et al. Estimation of effect size distribution from genome-wide association studies and implications for future discoveries. Nat Genet 2010; 42: 570-575.
    CrossrefPubMedGoogle Scholar
  • 15 Ross R. Mechanisms of disease: Atherosclerosis-An inflammatory disease. N Engl J Med 1999; 340: 115-126.
    CrossrefPubMedGoogle Scholar
  • 16 Lusis AJ. Atherosclerosis. Nature 2000; 407: 233-241.
    CrossrefPubMedGoogle Scholar
  • 17 Campbell JH, Campbell GR. Smooth Muscle Phenotypic Modulation—A Personal Experience. Arterioscler Thromb Vasc Biol 2012; 32: 1784-1789.
    CrossrefPubMedGoogle Scholar
  • 18 Gomez D, Owens GK. Smooth muscle cell phenotypic switching in atherosclerosis. Cardiovasc Res 2012; 95: 156-164.
    CrossrefPubMedGoogle Scholar
  • 19 Hao H. et al. Arterial smooth muscle cell heterogeneity: implications for atherosclerosis and restenosis development. Arterioscler Thromb Vasc Biol 2003; 23: 1510-1520.
    CrossrefPubMedGoogle Scholar
  • 20 Doran AC. et al. Role of smooth muscle cells in the initiation and early progression of atherosclerosis. Arterioscler Thromb Vasc Biol 2008; 28: 812-819.
    CrossrefPubMedGoogle Scholar
  • 21 Lacolley P. et al. The vascular smooth muscle cell in arterial pathology: a cell that can take on multiple roles. Cardiovasc Res 2012; 95: 194-204.
    CrossrefPubMedGoogle Scholar
  • 22 Johnson JL. Emerging regulators of vascular smooth muscle cell function in the development and progression of atherosclerosis. Cardiovasc Res. 2014 Epub ahead of print.
    PubMedGoogle Scholar
  • 23 Motterle A. et al. Functional analyses of coronary artery disease associated variation on chromosome 9p21 in vascular smooth muscle cells. Hum Mol Genet 2012; 21: 4021-4029.
    CrossrefPubMedGoogle Scholar
  • 24 Perisic L. et al. Profiling of atherosclerotic lesions by gene and tissue microar-rays reveals PCSK6 as a novel protease in unstable carotid atherosclerosis. Arte-rioscler Thromb Vasc Biol 2013; 33: 2432-2443.
    CrossrefPubMedGoogle Scholar
  • 25 Bozzini C. et al. Influence of polymorphisms in the factor VII gene promoter on activated factor VII levels and on the risk of myocardial infarction in advanced coronary atherosclerosis. Thromb Haemost 2004; 92: 541-549.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 26 Martinelli N. et al. Polymorphisms at LDLR locus may be associated with coronary artery disease through modulation of coagulation factor VIII activity and independently from lipid profile. Blood 2010; 116: 5688-5697.
    CrossrefPubMedGoogle Scholar
  • 27 Kathiresan S. Myocardial Infarction Genetics Consortium. Genome-wide association of early-onset myocardial infarction with single nucleotide polymorphisms and copy number variants. Nat Genet 2009; 41: 334-341.
    CrossrefPubMedGoogle Scholar
  • 28 Matthews DR. et al. Homeostasis model assessment: insulin resistance and beta-cell function from fasting plasma glucose and insulin concentrations in man. Diabetologia 1985; 28: 412-419.
    CrossrefPubMedGoogle Scholar
  • 29 North American Symptomatic Carotid EndarterectomyTrial Collaborators. Beneficial effect of carotid endarterectomy in symptomatic patients with highgrade carotid stenosis. N Engl J Med 1991; 325: 445-453.
    CrossrefPubMedGoogle Scholar
  • 30 Coen M. et al. Calmodulin expression distinguishes the smooth muscle cell population of human carotid plaque. Am J Pathol 2013; 183: 996-1009.
    CrossrefPubMedGoogle Scholar
  • 31 Skalli O. et al. A monoclonal antibody against alpha-smooth muscle actin: a new probe for smooth muscle differentiation. J Cell Biol 1986; 103: 2787-2796.
    CrossrefPubMedGoogle Scholar
  • 32 Cui X, Churchill GA. Statistical tests for differential expression in cDNA micro-array experiments. Genome Biol 2003; 4: 210-219.
    CrossrefPubMedGoogle Scholar
  • 33 Altman DG. Statistics and ethics in medical research. How large a sample? Br Med J 1980; 281: 1336-1338.
    PubMedGoogle Scholar
  • 34 Lake SL. et al. Estimation and tests of haplotype-environment interaction when linkage phase is ambiguous. Hum Hered 2003; 55: 56-65.
    CrossrefPubMedGoogle Scholar
  • 35 Park BY. et al. Differential parental transmission of markers in BCL3 among Korean cleft case-parent trios. J Prev Med Public Health 2009; 42: 1-4.
    CrossrefPubMedGoogle Scholar
  • 36 Reilly MP. et al. Myocardial Infarction Genetics Consortium; Wellcome Trust Case Control Consortium. Identification of ADAMTS7 as a novel locus for coronary atherosclerosis and association of ABO with myocardial infarction in the presence of coronary atherosclerosis: two genome-wide association studies. Lancet 2011; 377: 383-392.
    CrossrefPubMedGoogle Scholar
  • 37 Folkersen L. et al. BiKE and ASAP study groups. Association of genetic risk variants with expression of proximal genes identifies novel susceptibility genes for cardiovascular disease. Circ Cardiovasc Genet 2010; 3: 365-373.
    CrossrefPubMedGoogle Scholar
  • 38 Ramsey SA. et al. A systems biology approach to understanding atherosclerosis. EMBO Mol Med 2010; 2: 79-89.
    CrossrefPubMedGoogle Scholar
  • 39 Maouche S, Schunkert H. Strategies beyond genome-wide association studies for atherosclerosis. Arterioscler Thromb Vasc Biol 2012; 32: 170-181.
    CrossrefPubMedGoogle Scholar
  • 40 Oram JF, Vaughan AM. ATP-Binding cassette cholesterol transporters and cardiovascular disease. Circ Res 2006; 99: 1031-1043.
    CrossrefPubMedGoogle Scholar
  • 41 Fitzgerald ML. et al. ABC transporters, atherosclerosis and inflammation. Atherosclerosis 2010; 211: 361-370.
    CrossrefPubMedGoogle Scholar
  • 42 Talmud PJ. et al. Gene-centric association signals for lipids and apolipoproteins identified via the Human CVD BeadChip. Am J Hum Genet 2009; 85: 628-642.
    CrossrefPubMedGoogle Scholar
  • 43 Fransen K. et al. Analysis of SNPs with an effect on gene expression identifies UBE2L3 and BCL3 as potential new risk genes for Crohn’s disease. Hum Mol Genet 2010; 19: 3482-3488.
    CrossrefPubMedGoogle Scholar
  • 44 Miller AM, McInnes IB. Cytokines as therapeutic targets to reduce cardiovascular risk in chronic inflammation. Curr Pharm Des 2011; 17: 1-8.
    CrossrefPubMedGoogle Scholar
  • 45 Palmer S, Chen YH. Bcl-3, a multifaced modulator of NF-kB-mediated gene transcription. Immunol Res 2008; 42: 210-218.
    CrossrefPubMedGoogle Scholar
  • 46 Hinz M. et al. It takes two to tango: IkBs, the multifunctional partners of NF-kB. Immunol Rev 2012; 246: 59-76.
    CrossrefPubMedGoogle Scholar
  • 47 Yang J. et al. BCL3 interacts cooperatively with peroxisome proliferator-activated receptor gamma (PPARgamma) coactivator 1alpha to coactivate nuclear receptors estrogen-related receptor alpha and PPARalpha. Mol Cell Biol 2009; 29: 4091-4102.
    CrossrefPubMedGoogle Scholar
  • 48 Alberti KG. et al. Harmonizing the metabolic syndrome: a joint interim statement of the International Diabetes Federation Task Force on Epidemiology and Prevention; National Heart, Lung, and Blood Institute; American Heart Association; World Heart Federation; International Atherosclerosis Society; and International Association for the Study of Obesity. Circulation 2009; 120: 1640-1645.
    CrossrefPubMedGoogle Scholar
  • 49 Grundy SM. Metabolic syndrome: a multiplex cardiovascular risk factor. J Clin Endocrinol Metab 2007; 92: 399-404.
    CrossrefPubMedGoogle Scholar
  • 50 Executive Summary of The Third Report of The National Cholesterol Education Program (NCEP) Expert Panel on Detection, Evaluation, And Treatment of High Blood Cholesterol In Adults (Adult Treatment Panel III). J Am Med Assoc 2001; 285: 2486-2497.
    CrossrefPubMedGoogle Scholar