Antiphospholipid antibodies in a large population-based cohort: genome-wide associations and effects on monocyte gene expression

Nadine Müller-Calleja; Heidi Rossmann; Christian Müller; Philipp Wild; Stefan Blankenberg; Norbert Pfeiffer; Harald Binder; Manfred E. Beutel; Davit Manukyan; Tanja Zeller; Karl J. Lackner

doi:10.1160/TH15-12-0947

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2016; 116(01): 115-123
DOI: 10.1160/TH15-12-0947

Blood Cells, Inflammation and Infection

Schattauer GmbH

Antiphospholipid antibodies in a large population-based cohort: genome-wide associations and effects on monocyte gene expression

Authors

Nadine Müller-Calleja^*

¹Institute of Clinical Chemistry and Laboratory Medicine, University Medical Center Mainz, Mainz, Germany
Heidi Rossmann^*

¹Institute of Clinical Chemistry and Laboratory Medicine, University Medical Center Mainz, Mainz, Germany
Christian Müller^*

²Clinic of General and Interventional Cardiology, University Heart Center Hamburg-Eppendorf, Hamburg, Germany

³Deutsches Zentrum für Herzkreislaufforschung (DZHK), Partner Site Hamburg Lübeck Kiel, Hamburg, Germany
Philipp Wild

⁴Department of Medicine 2, University Medical Center Mainz, Mainz, Germany

⁵Deutsches Zentrum für Herzkreislaufforschung (DZHK), partner site RhineMain, Mainz, Germany

⁶Center for Thrombosis and Hemostasis, University Medical Center Mainz, Mainz, Germany
Stefan Blankenberg

²Clinic of General and Interventional Cardiology, University Heart Center Hamburg-Eppendorf, Hamburg, Germany

³Deutsches Zentrum für Herzkreislaufforschung (DZHK), Partner Site Hamburg Lübeck Kiel, Hamburg, Germany
Norbert Pfeiffer

⁷Department of Ophthalmology, University Medical Center Mainz, Mainz, Germany
Harald Binder

⁸Institute of Medical Biostatistics, Epidemiology and Informatics, University Medical Center Mainz, Mainz Germany
Manfred E. Beutel

⁹Department of Psychosomatic Medicine and Psychotherapy, University Medical Center Mainz, Mainz, Germany
Davit Manukyan

¹Institute of Clinical Chemistry and Laboratory Medicine, University Medical Center Mainz, Mainz, Germany
Tanja Zeller

²Clinic of General and Interventional Cardiology, University Heart Center Hamburg-Eppendorf, Hamburg, Germany

³Deutsches Zentrum für Herzkreislaufforschung (DZHK), Partner Site Hamburg Lübeck Kiel, Hamburg, Germany
Karl J. Lackner

¹Institute of Clinical Chemistry and Laboratory Medicine, University Medical Center Mainz, Mainz, Germany

Abstract

Summary

The antiphospholipid syndrome (APS) is characterised by venous and/ or arterial thrombosis and pregnancy morbidity in women combined with the persistent presence of antiphospholipid antibodies (aPL). We aimed to identify genetic factors associated with the presence of aPL in a population based cohort. Furthermore, we wanted to clarify if the presence of aPL affects gene expression in circulating monocytes. Titres of IgG and IgM against cardiolipin, D2glycoprotein 1 (antiD2GPI), and IgG against domain 1 of D2GPI (anti-domain 1) were determined in approx. 5,000 individuals from the Gutenberg Health Study (GHS) a population based cohort of German descent. Genotyping was conducted on Affymetrix Genome-Wide Human SNP 6.0 arrays. Monocyte gene expression was determined in a subgroup of 1,279 individuals by using the Illumina HT-12 v3 BeadChip. Gene expression data were confirmed in vitro and ex vivo by qRT-PCR. Genome wide analysis revealed significant associations of anti-D2GPI IgG and APOH on chromosome 17, which had been previously identified by candidate gene approaches, and of anti-domain1 and MACROD2 on chromosome 20 which has been listed in a previous GWAS as a suggestive locus associated with the occurrence of anti-D2GPI antibodies. Expression analysis confirmed increased expression of TNFD in monocytes and identified and confirmed neuron navigator 3 (NAV3) as a novel gene induced by aPL. In conclusion, MACROD2 represents a novel genetic locus associated with aPL. Furthermore, we show that aPL induce the expression of NAV3 in monocytes and endothelial cells. This will stimulate further research into the role of these genes in the APS.

Keywords

Antiphospholipid antibodies - genome-wide association study - gene expression - monocytes - cohort study

Full Text

References

References
1 Wilson WA, Gharavi AE, Koike T. et al. International consensus statement on preliminary classification criteria for definite antiphospholipid syndrome: report of an international workshop. Arthritis Rheum 1999; 42: 1309-1311.
2 Miyakis S, Lockshin MD, Atsumi T. et al. International consensus statement on an update of the classification criteria for definite antiphospholipid syndrome (APS). J Thromb Haemost 2006; 04: 295-306.
3 Lackner KJ, Peetz D, von Landenberg P. Revision of the Sapporo criteria for the antiphospho-lipid syndrome - Coming to grips with evidence and Thomas Bayes?. Thromb Haemost 2006; 95: 917-919.
4 Galli M, Luciani D, Bertolini G. et al. Lupus anticoagulants are stronger risk factors for thrombosis than anticardiolipin antibodies in the antiphospholipid syndrome: a systematic review of the literature. Blood 2003; 101: 1827-1832.
5 Galli M, Luciani D, Bertolini G. et al. Anti-beta 2-glycoprotein I, antiprothrombin antibodies, and the risk of thrombosis in the antiphospholipid syndrome. Blood 2003; 102: 2717-2723.
6 Reynaud Q, Lega J-C, Mismetti P. et al. Risk of venous and arterial thrombosis according to type of antiphospholipid antibodies in adults without systemic lupus erythematosus: A systematic review and meta-analysis. Autoimmun Rev 2014; 13: 595-608.
7 Meroni PL, Borghi MO, Raschi E. et al. Pathogenesis of the antiphospholipid syndrome: understanding the antibodies. Nat Rev Rheumatol 2011; 07: 330-339.
8 Willis R, Harris EN, Pierangeli SS. Pathogenesis of the antiphospholipid syndrome. Semin Thromb Hemost 2012; 38: 305-321.
9 Giannakopoulos B, Krilis SA. The pathogenesis of the antiphospholipid syndrome. N Engl J Med 2013; 368: 1033-1044.
10 de Groot PG, Urbanus RT, Derksen RH. Pathophysiology of thrombotic APS: where do we stand?. Lupus 2012; 21: 704-707.
11 Kamboh MI, Wang X, Kao AH. et al. Genome-wide association study of antiphospholipid antibodies. Autoimmune Dis 2013; 2013: 761046.
12 Prinz N, Clemens N, Canisius A. et al. Endosomal NADPH-oxidase is critical for induction of the tissue factor gene in monocytes and endothelial cells. Lessons from the antiphospholipid syndrome. Thromb Haemost 2013; 109: 525-531.
13 Müller-Calleja N, Köhler A, Siebald B. et al. Cofactor-independent antiphospholipid antibodies activate the NLRP3-inflammasome via endosomal NADPHoxidase: implications for the antiphospholipid syndrome. Thromb Haemost 2015; 113: 1071-1083.
14 Wild PS, Zeller T, Beutel M. et al. The Gutenberg Health Study. Bundesgesundheitsblatt Gesundheitsforschung Gesundheitsschutz 2012; 55: 824-829.
15 Rotival M, Zeller T, Wild PS. et al. Integrating genome-wide genetic variations and monocyte expression data reveals trans-regulated gene modules in humans. PLoS Genet 2011; 07: e1002367.
16 Zeller T, Wild P, Szymczak S. et al. Genetics and beyond--the transcriptome of human monocytes and disease susceptibility. PLoS One 2010; 05: e10693.
17 Howie B, Fuchsberger C, Stephens M. et al. Fast and accurate genotype imputation in genome-wide association studies through pre-phasing. Nat Genet 2012; 44: 955-959.
18 Schurmann C, Heim K, Schillert A. et al. Analyzing illumina gene expression microarray data from different tissues: methodological aspects of data analysis in the metaxpress consortium. PLoS One 2012; 07: e50938.
19 Aulchenko YS, Struchalin MV, van Duijn CM. ProbABEL package for genomewide association analysis of imputed data. BMC Bioinformatics 2010; 11: 134.
20 Willer CJ, Li Y, Abecasis GR. METAL: fast and efficient meta-analysis of genomewide association scans. Bioinformatics 2010; 26: 2190-2191.
21 von Landenberg C, Lackner KJ, von Landenberg P. et al. Isolation and characterisation of two human monoclonal anti-phospholipid IgG from patients with autoimmune disease. J Autoimmun 1999; 13: 215-223.
22 Döring Y, Hurst J, Lorenz M, Prinz N. et al. Human antiphospholipid antibodies induce TNFalpha in monocytes via Toll-like receptor 8. Immunobiology 2010; 215: 230-241.
23 Chamorro AJ, Marcos M, Mirón-Canelo JA. et al. Val247Leu β2-glycoprotein-I allelic variant is associated with antiphospholipid syndrome: systematic review and meta-analysis. Autoimmun Rev 2012; 11: 705-712.
24 Yasuda S, Atsumi T, Matsuura E. et al. Significance of valine/leucine247 polymorphism of beta2-glycoprotein I in antiphospholipid syndrome: increased reactivity of anti-beta2-glycoprotein I autoantibodies to the valine247 beta2-glycoprotein I variant. Arthritis Rheum 2005; 52: 212-218.
25 Hirose N, Williams R, Alberts AR. et al. A role for the polymorphism at position 247 of the beta2-glycoprotein I gene in the generation of anti-beta2-glycoprotein I antibodies in the antiphospholipid syndrome. Arthritis Rheum 1999; 42: 1655-1661.
26 Han W, Li X, Fu X. The macro domain protein family: structure, function, and their potential therapeutic implications. Mut Res 2011; 727: 86-103.
27 Jones RM, Cadby G, Blangero J. et al. MACROD2 gene associated with autisticlike traits in a general population sample. Psychiatr Genet 2014; 24: 241-248.
28 Wang YT, Sung PY, Lin PL. et al. A multi-SNP association test for complex dieseases incorporating an optimal P-value threshold algorithm in nuclear families. BMC Genomics 2015; 16: 381.
29 Careaga M, Hansen RL, Hertz-Piccotto I. et al. Increased anti-phospholipid antibodies in autism spectrum disorders. Mediators Inflamm 2013; 2013: 935608.
30 Abisror N, Mekinian A, Lachassinne E. et al. Autism spectrum disorders in babies born to mothers with antiphospholipid syndrome. Semin Arthritis Rheum 2013; 43: 348-351.
31 Holdt LM, Hoffmann S, Sass K. et al. Alu elements in ANRIL non-coding RNA at chromosome 9p21 modulate atherogenic cell functions through trans-regulation of gene networks. PLoS Genet 2013; 09: e1003588.
32 Prinz N, Clemens N, Strand D. et al. Antiphospholipid-antibodies induce translocation of TLR7 and TLR8 to the endosome in human monocytes and plasmacytoid dendritic cells. Blood 2011; 118: 2322-2332.
33 Coy JF, Wiemann S, Bechmann I. et al. Pore membrane and/or filament interacting like protein 1 (POMFIL1) is predominantly expressed in the nervous system and encodes different protein isoforms. Gene 2002; 290: 73-94.
34 Maes T, Barceló A, Buesa C. Neuron navigator: a human gene family with homology to unc-53, a cell guidance gene from Caenorhabditis elegans. Genomics 2002; 80: 21-30.
35 Cohen-Dvashi H, Ben-Chetrit N, Russell R. et al. Navigator-3, a modulator of cell migration, may act as a suppressor of breast cancer progression. EMBO Mol Med 2015; 07: 299-314.
36 Karenko L, Hahtola S, Päivinen S. et al. Primary cutaneous T-cell lymphomas show a deletion or translocation affecting NAV3, the human UNC-53 homologue. Cancer Res 2005; 65: 8101-8110.
37 Hahtola S, Burghart E, Puputti M. et al. Cutaneous T-cell lymphoma-associated lung cancers show chromosomal aberrations differing from primary lung cancer. Genes Chromosomes Cancer 2008; 47: 107-117.
38 Carlsson E, Ranki A, Sipilä L. et al. Potential role of a navigator gene NAV3 in colorectal cancer. Br J Cancer 2012; 106: 517-524.

Supplementary Material

Online Supplementary Material (PDF) (PDF)