Tag Variants of LGALS-3 Containing Haplotype Block in Advanced Carotid Atherosclerosis

  • Ana Djordjevic
    Corresponding author.
    Department of Radiobiology and Molecular Genetics, “Vinca” Institute of Nuclear Sciences-National Institute of the Republic of Serbia, Mike Petrovica Alasa 12-14, P.O. Box 522, University of Belgrade, Belgrade 11001, Serbia
    Search for articles by this author
  • Maja Zivkovic
    Department of Radiobiology and Molecular Genetics, “Vinca” Institute of Nuclear Sciences-National Institute of the Republic of Serbia, Mike Petrovica Alasa 12-14, P.O. Box 522, University of Belgrade, Belgrade 11001, Serbia
    Search for articles by this author
  • Igor Koncar
    Clinic for Vascular and Endovascular Surgery, Clinical Center of Serbia, Belgrade, Serbia

    Medical Faculty, University of Belgrade, Belgrade, Serbia
    Search for articles by this author
  • Aleksandra Stankovic
    Department of Radiobiology and Molecular Genetics, “Vinca” Institute of Nuclear Sciences-National Institute of the Republic of Serbia, Mike Petrovica Alasa 12-14, P.O. Box 522, University of Belgrade, Belgrade 11001, Serbia
    Search for articles by this author
  • Jovana Kuveljic
    Department of Radiobiology and Molecular Genetics, “Vinca” Institute of Nuclear Sciences-National Institute of the Republic of Serbia, Mike Petrovica Alasa 12-14, P.O. Box 522, University of Belgrade, Belgrade 11001, Serbia
    Search for articles by this author
  • Tamara Djuric
    Department of Radiobiology and Molecular Genetics, “Vinca” Institute of Nuclear Sciences-National Institute of the Republic of Serbia, Mike Petrovica Alasa 12-14, P.O. Box 522, University of Belgrade, Belgrade 11001, Serbia
    Search for articles by this author



      Galectin-3 affects a variety of biological processes. It is encoded by LGALS-3, located in unique haplotype block in Caucasians. Most of the studies regarding the gal-3 role in atherosclerosis are focused exclusively on protein/mRNA levels. Genetic analyses of LGALS-3 are scarce. We sought to thoroughly examine the genetic background of gal-3 and to analyze tag variants that cover more than 80% variability of the LGALS-3 containing hap-block in association with carotid plaque presence (CPP). According to Tagger server, rs4040064 G/T, rs11628437 G/A and rs7159490 C/T cover 82% (r2 > 0.8) of the genetic variance of this hap-block. Our aims were to investigate possible association of rs4040064, rs11628437 and rs7159490 haplotypes with CPP in patients with advanced carotid atherosclerosis (CA) and to analyze their possible effect on LGALS-3 mRNA expression in carotid plaques.

      Materials and methods

      Study group consisted of 468 patients and 296 controls. Rs4040064, rs11628437, rs7159490 and LGALS-3 mRNA expression were detected by TaqMan® technology.


      We have found that haplotype TAC was associated with the cerebrovascular insult (CVI) occurrence (OR = 1.68, 95% CI = 1.09-2.58, p = 0.02), compared to the referent haplotype. OR was adjusted for hypertension, age and BMI. TAC also showed higher, but not statistically significant, LGALS-3 expression in carotid plaques.


      Our results suggest that rs4040064, rs11628437 and rs7159490 bear no association with CPP, neither they affect LGALS-3 mRNA in carotid plaques. However, we showed a significant association of haplotype TAC with the CVI occurrence in CA patients from Serbia. Replication and validation of our results are required.

      Key Words

      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 to Journal of Stroke and Cerebrovascular Diseases
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect


        • Falk E.
        Pathogenesis of atherosclerosis.
        J Am Coll Cardiol. 2006; 47: C7-12
        • Hansson G.K.
        • Hermansson A.
        The immune system in atherosclerosis.
        Nat Immunol. 2011; 12: 204-212
        • Ross R.
        Atherosclerosis–an inflammatory disease.
        N Engl J Med. 1999; 340: 115-126
        • Leffler H.
        • Carlsson S.
        • Hedlund M.
        • et al.
        Introduction to galectins.
        Glycoconj J. 2002; 19: 433-440
        • Seetharaman J.
        • Kanigsberg A.
        • Slaaby R.
        • et al.
        X-ray crystal structure of the human galectin-3 carbohydrate recognition domain at 2.1-A resolution.
        J Biol Chem. 1998; 273: 13047-13052
        • Di Lella S.
        • Sundblad V.
        • Cerliani J.P.
        • et al.
        When galectins recognize glycans: from biochemistry to physiology and back again.
        Biochemistry. 2011; 50: 7842-7857
        • Hsu D.K.
        • Zuberi R.I.
        • Liu FT.
        Biochemical and biophysical characterization of human recombinant IgE-binding protein, an S-type animal lectin.
        J Biol Chem. 1992; 267: 14167-14174
        • Liu F.T.
        • Rabinovich GA.
        Galectins as modulators of tumour progression.
        Nat Rev Cancer. 2005; 5: 29-41
        • Lok D.J.
        • Lok S.I.
        • Bruggink-André de la Porte P.W.
        • et al.
        Galectin-3 is an independent marker for ventricular remodeling and mortality in patients with chronic heart failure.
        Clin Res Cardiol. 2013; 102: 103-110
        • Ohshima S.
        • Kuchen S.
        • Seemayer C.A.
        • et al.
        Galectin 3 and its binding protein in rheumatoid arthritis.
        Arthritis Rheum. 2003; 48: 2788-2795
        • Ezzat M.H.
        • El-Gammasy T.M.
        • Shaheen K.Y.
        • et al.
        Elevated production of galectin-3 is correlated with juvenile idiopathic arthritis disease activity, severity, and progression.
        Int J Rheum Dis. 2011; 14: 345-352
        • Iacovazzi P.A.
        • Notarnicola M.
        • Caruso M.G.
        • et al.
        Serum levels of galectin-3 and its ligand 90k/mac-2bp in colorectal cancer patients.
        Immunopharmacol Immunotoxicol. 2010; 32: 160-164
        • Kadoglou N.P.
        • Sfyroeras G.S.
        • Spathis A.
        • et al.
        Galectin-3, carotid plaque vulnerability, and potential effects of statin therapy.
        Eur J Vasc Endovasc Surg. 2015; 49: 4-9
        • Iacobini C.
        • Menini S.
        • Ricci C.
        • et al.
        Accelerated lipid-induced atherogenesis in galectin-3-deficient mice: role of lipoxidation via receptor-mediated mechanisms.
        Arterioscler Thromb Vasc Biol. 2009; 29: 831-836
        • Menini S.
        • Iacobini C.
        • Ricci C.
        • et al.
        The galectin-3/RAGE dyad modulates vascular osteogenesis in atherosclerosis.
        Cardiovasc Res. 2013; 100: 472-480
        • Papaspyridonos M.
        • McNeill E.
        • de Bono J.P.
        • et al.
        Galectin-3 is an amplifier of inflammation in atherosclerotic plaque progression through macrophage activation and monocyte chemoattraction.
        Arterioscler Thromb Vasc Biol. 2008; 28: 433-440
        • Djordjevic A.
        • Zivkovic M.
        • Stankovic A.
        • et al.
        Genetic variants in the vicinity of LGALS-3 gene and LGALS-3 mRNA expression in advanced carotid atherosclerosis: an exploratory study.
        J Clin Lab Anal. 2016; 30: 1150-1157
        • de Boer R.A.
        • Verweij N.
        • van Veldhuisen D.J.
        • et al.
        A genome-wide association study of circulating galectin-3.
        PLoS One. 2012; 7: e47385
        • Boyle A.P.
        • Hong E.L.
        • Hariharan M.
        • et al.
        Annotation of functional variation in personal genomes using RegulomeDB.
        Genome Res. 2012; 22: 1790-1797
        • de Bakker P.I.
        • Yelensky R.
        • Pe'er I.
        • et al.
        Efficiency and power in genetic association studies.
        Nat Genet. 2005; 37: 1217-1223
        • Barnett H.J.M.
        • Taylor D.W.
        • Haynes R.B.
        • et al.
        Beneficial effect of carotid endarterectomy in symptomatic patients with high-grade carotid stenosis.
        N Engl J Med. 1991; 325: 445-453
        • Gray-Weale A.C.
        • Graham J.C.
        • Burnett J.R.
        • et al.
        Carotid artery atheroma: comparison of preoperative B-mode ultrasound appearance with carotid endarterectomy specimen pathology.
        J Cardiovasc Surg (Torino). 1988; 29: 676-681
        • Kunkel L.M.
        • Smith K.D.
        • Boyer S.H.
        • et al.
        Analysis of human Y-chromosome-specific reiterated DNA in chromosome variants.
        Proc Natl Acad Sci USA. 1977; 74: 1245-1249
        • Tregouet D.A.
        • Escolano S.
        • Tiret L.
        • et al.
        A new algorithm for haplotype-based association analysis: the stochastic-EM algorithm.
        Ann Hum Genet. 2004; 68: 165-177
        • DA T.
        • Garelle V.
        A new JAVA interface implementation of THESIAS: testing haplotype effects in association studies.
        Bioinformatics. 2007; 23: 1038-1039
        • Dupont W.D.
        • Jr P.W.D.
        Power and sample size calculations. A review and computer program.
        Control Clin Trials. 1990; 11: 116-128
        • Pfaffl M.W.
        • Horgan G.W.
        • Dempfle L.
        Relative expression software tool (REST) for group-wise comparison and statistical analysis of relative expression results in real-time PCR.
        Nucleic Acids Res. 2002; 30: e36
        • Cruz Gda S.
        • Angelo A.L.
        • Larocca T.F.
        • et al.
        Assessment of galectin-3 polymorphism in subjects with chronic Chagas disease.
        Arq Bras Cardiol. 2015; 105: 472-478
        • Wu F.
        • Hu N.
        • Li Y.
        • et al.
        Galectin-3 genetic variants are associated with platinum-based chemotherapy response and prognosis in patients with NSCLC.
        Cell Oncol. 2012; 35 (Dordr): 175-180
        • Chen H.J.
        • Zheng Z.C.
        • Yuan B.Q.
        • et al.
        The effect of galectin-3 genetic variants on the susceptibility and prognosis of gliomas in a Chinese population.
        Neurosci Lett. 2012; 518: 1-4
        • Trompet S.
        • Jukema W.
        • Mooijaart S.P.
        • et al.
        Genetic variation in galectin-3 gene associates with cognitive function at old age.
        Neurobiol Aging. 2012; 33: 2232.e1-2232.e9
        • Hu C.Y.
        • Chang S.K.
        • Wu C.S.
        • et al.
        Galectin-3 gene (LGALS3) +292C allele is a genetic predisposition factor for rheumatoid arthritis in Taiwan.
        Clin Rheumatol. 2011; 30: 1227-1233
        • Vuckovic D.
        • Bao E.L.
        • Akbari P.
        • et al.
        The polygenic and monogenic basis of blood traits and diseases.
        Cell. 2020; 182 (e11): 1214-1231
        • Lam M.
        • Hill W.D.
        • Trampush J.W.
        • et al.
        Pleiotropic meta-analysis of cognition, education, and schizophrenia differentiates roles of early neurodevelopmental and adult synaptic pathways.
        Am J Hum Genet. 2019; 105: 334-350
        • Okamura K.
        • Abe Y.
        • Naka I.
        • et al.
        Genome-wide association study identifies CDH13 as a susceptibility gene for rhododendrol-induced leukoderma.
        Pigment Cell Melanoma Res. 2020; 33: 826-833
        • Folkersen L.
        • Gustafsson S.
        • Wang Q.
        • et al.
        Genomic and drug target evaluation of 90 cardiovascular proteins in 30,931 individuals.
        Nat Metab. 2020; 2: 1135-1148
        • Mitchell B.D.
        • Fornage M.
        • McArdle P.F.
        • et al.
        Using previously genotyped controls in genome-wide association studies (GWAS): application to the stroke genetics network (SiGN).
        Front Genet. 2014; 5: 95
        • Carty C.L.
        • Keene K.L.
        • Cheng Y.C.
        • et al.
        Meta-analysis of genome-wide association studies identifies genetic risk factors for stroke in African Americans.
        Stroke. 2015; 46: 2063-2068
        • Rasmussen-Torvik L.J.
        • Pacheco J.A.
        • Wilke R.A.
        • et al.
        High density GWAS for LDL cholesterol in African Americans using electronic medical records reveals a strong protective variant in APOE.
        Clin Transl Sci. 2012; 5: 394-399
        • Frau F.
        • Zaninello R.
        • Salvi E.
        • et al.
        Genome-wide association study identifies CAMKID variants involved in blood pressure response to losartan: the SOPHIA study.
        Pharmacogenomics. 2014; 15: 1643-1652
        • Karczewski K.J.
        • Francioli L.C.
        • Tiao G.
        • et al.
        The mutational constraint spectrum quantified from variation in 141,456 humans.
        Nature. 2020; 581: 434-443
        • Böhm B.
        • Hartmann K.
        • Buck M.
        • et al.
        Sex differences of carotid intima-media thickness in healthy children and adolescents.
        Atherosclerosis. 2009; 206: 458-463
        • Juonala M.
        • Kähönen M.
        • Laitinen T.
        • et al.
        Effect of age and sex on carotid intima-media thickness, elasticity and brachial endothelial function in healthy adults: the cardiovascular risk in Young Finns Study.
        Eur Heart J. 2008; 29: 1198-1206
        • Halvorsen D.S.
        • Johnsen S.H.
        • Mathiesen E.B.
        • et al.
        The association between inflammatory markers and carotid atherosclerosis is sex dependent: the Tromsø Study.
        Cerebrovasc Dis. 2009; 27: 392-397
        • Sinning C.
        • Wild P.S.
        • Echevarria F.M.
        • et al.
        Sex differences in early carotid atherosclerosis (from the community-based Gutenberg-heart study).
        Am J Cardiol. 2011; 107: 1841-1847
        • Kaur T.
        • Thakur K.
        • Singh J.
        • et al.
        Genotypic-phenotypic screening of galectin-3 in relation to risk towards rheumatoid arthritis.
        Arch Med Res. 2019; 50: 214-224
        • Ward L.D.
        • Kellis M.
        HaploReg: a resource for exploring chromatin states, conservation, and regulatory motif alterations within sets of genetically linked variants.
        Nucleic Acids Res. 2012; 40: D930-D934
        • Zhang Y.
        • Wang Y.
        • Zhai M.
        • et al.
        Influence of LGALS3 gene polymorphisms on susceptibility and prognosis of dilated cardiomyopathy in a Northern Han Chinese population.
        Gene. 2018; 642: 293-298
        • Djordjevic A.
        • Dekleva M.
        • Zivkovic M.
        • et al.
        Left ventricular remodeling after the first myocardial infarction in association with LGALS-3 neighbouring variants rs2274273 and rs17128183 and its relative mRNA expression: a prospective study.
        Mol Biol Rep. 2018; 45: 2227-2236
        • Zaborska B.
        • Sygitowicz G.
        • Smarż K.
        • et al.
        Galectin-3 is related to right ventricular dysfunction in heart failure patients with reduced ejection fraction and may affect exercise capacity.
        Sci Rep. 2020; 10: 16682
        • Ho J.E.
        • Liu C.
        • Lyass A.
        • Courchesne P.
        • et al.
        Galectin-3, a marker of cardiac fibrosis, predicts incident heart failure in the community.
        J Am Coll Cardiol. 2012; 60: 1249-1256
        • Felker G.M.
        • Fiuzat M.
        • Shaw L.K.
        • et al.
        Galectin-3 in ambulatory patients with heart failure: results from the HF-ACTION study.
        Circ Heart Fail. 2012; 5: 72-78
        • Chen A.
        • Hou W.
        • Zhang Y.
        • et al.
        Prognostic value of serum galectin-3 in patients with heart failure: a meta-analysis.
        Int J Cardiol. 2015; 182: 168-170
        • Madrigal-Matute J.
        • Lindholt J.S.
        • Fernandez-Garcia C.E.
        • et al.
        Galectin-3, a biomarker linking oxidative stress and inflammation with the clinical outcomes of patients with atherothrombosis.
        J Am Heart Assoc. 2014; 3e000785
        • de Boer R.A.
        • van Veldhuisen D.J.
        • Gansevoort R.T.
        • et al.
        The fibrosis marker galectin-3 and outcome in the general population.
        J Intern Med. 2012; 272: 55-64
        • Edsfeldt A.
        • Bengtsson E.
        • Asciutto G.
        • et al.
        High plasma levels of galectin-3 are associated with increased risk for stroke after carotid endarterectomy.
        Cerebrovasc Dis. 2016; 41: 199-203
        • He X.W.
        • Li W.L.
        • Li C.
        • et al.
        Serum levels of galectin-1, galectin-3, and galectin-9 are associated with large artery atherosclerotic stroke.
        Sci Rep. 2017; 7: 40994
        • O'Donnell C.J.
        • Kavousi M.
        • Smith A.V.
        • et al.
        Genome-wide association study for coronary artery calcification with follow-up in myocardial infarction.
        Circulation. 2011; 124: 2855-2864
        • Emilsson V.
        • Ilkov M.
        • Lamb J.R.
        • et al.
        Co-regulatory networks of human serum proteins link genetics to disease.
        Science. 2018; 361: 769-773