Preferential association of a functional variant in complement receptor 2 with antibodies to double-stranded DNA

BIOLUPUS and GENLES Networks

Research output: Contribution to journalArticle

Abstract

Objectives Systemic lupus erythematosus (SLE; OMIM 152700) is characterised by the production of antibodies to nuclear antigens. We previously identified variants in complement receptor 2 (CR2/CD21) that were associated with decreased risk of SLE. This study aimed to identify the causal variant for this association. Methods Genotyped and imputed genetic variants spanning CR2 were assessed for association with SLE in 15 750 case-control subjects from four ancestral groups. Allele-specific functional effects of associated variants were determined using quantitative real-time PCR, quantitative flow cytometry, electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation (ChIP)-PCR. Results The strongest association signal was detected at rs1876453 in intron 1 of CR2 (pmeta=4.2×10-4, OR 0.85), specifically when subjects were stratified based on the presence of dsDNA autoantibodies (case-control pmeta=7.6×10-7, OR 0.71; case-only pmeta=1.9×10-4, OR 0.75). Although allele-specific effects on B cell CR2 mRNA or protein levels were not identified, levels of complement receptor 1 (CR1/CD35) mRNA and protein were significantly higher on B cells of subjects harbouring the minor allele (p=0.0248 and p=0.0006, respectively). The minor allele altered the formation of several DNA protein complexes by EMSA, including one containing CCCTC-binding factor (CTCF), an effect that was confirmed by ChIP-PCR. Conclusions These data suggest that rs1876453 in CR2 has long-range effects on gene regulation that decrease susceptibility to lupus. Since the minor allele at rs1876453 is preferentially associated with reduced risk of the highly specific dsDNA autoantibodies that are present in preclinical, active and severe lupus, understanding its mechanisms will have important therapeutic implications.

Original languageEnglish (US)
Pages (from-to)242-252
Number of pages11
JournalAnnals of the Rheumatic Diseases
Volume75
Issue number1
DOIs
StatePublished - Jan 1 2016

Fingerprint

Complement 3d Receptors
Electrophoretic mobility
Alleles
Association reactions
Autoantibodies
Chromatin
Antibodies
Assays
DNA
Cells
Chromatin Immunoprecipitation
Complement Receptors
Electrophoretic Mobility Shift Assay
Nuclear Antigens
Messenger RNA
Proteins
Flow cytometry
Gene expression
B-Lymphocytes
Introns

ASJC Scopus subject areas

  • Rheumatology
  • Immunology
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Allergy

Cite this

Preferential association of a functional variant in complement receptor 2 with antibodies to double-stranded DNA. / BIOLUPUS and GENLES Networks.

In: Annals of the Rheumatic Diseases, Vol. 75, No. 1, 01.01.2016, p. 242-252.

Research output: Contribution to journalArticle

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title = "Preferential association of a functional variant in complement receptor 2 with antibodies to double-stranded DNA",
abstract = "Objectives Systemic lupus erythematosus (SLE; OMIM 152700) is characterised by the production of antibodies to nuclear antigens. We previously identified variants in complement receptor 2 (CR2/CD21) that were associated with decreased risk of SLE. This study aimed to identify the causal variant for this association. Methods Genotyped and imputed genetic variants spanning CR2 were assessed for association with SLE in 15 750 case-control subjects from four ancestral groups. Allele-specific functional effects of associated variants were determined using quantitative real-time PCR, quantitative flow cytometry, electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation (ChIP)-PCR. Results The strongest association signal was detected at rs1876453 in intron 1 of CR2 (pmeta=4.2×10-4, OR 0.85), specifically when subjects were stratified based on the presence of dsDNA autoantibodies (case-control pmeta=7.6×10-7, OR 0.71; case-only pmeta=1.9×10-4, OR 0.75). Although allele-specific effects on B cell CR2 mRNA or protein levels were not identified, levels of complement receptor 1 (CR1/CD35) mRNA and protein were significantly higher on B cells of subjects harbouring the minor allele (p=0.0248 and p=0.0006, respectively). The minor allele altered the formation of several DNA protein complexes by EMSA, including one containing CCCTC-binding factor (CTCF), an effect that was confirmed by ChIP-PCR. Conclusions These data suggest that rs1876453 in CR2 has long-range effects on gene regulation that decrease susceptibility to lupus. Since the minor allele at rs1876453 is preferentially associated with reduced risk of the highly specific dsDNA autoantibodies that are present in preclinical, active and severe lupus, understanding its mechanisms will have important therapeutic implications.",
author = "{BIOLUPUS and GENLES Networks} and Jian Zhao and Giles, {Brendan M.} and Taylor, {Rhonda L.} and Yette, {Gabriel A.} and Lough, {Kara M.} and Ng, {Han Leng} and Abraham, {Lawrence J.} and Hui Wu and Kelly, {Jennifer A.} and Glenn, {Stuart B.} and Adler, {Adam J.} and Williams, {Adrienne H.} and Comeau, {Mary E.} and Ziegler, {Julie T.} and Miranda Marion and Alarc{\'o}n-Riquelme, {Marta E.} and Alarc{\'o}n, {Graciela S.} and Anaya, {Juan Manuel} and Bae, {Sang Cheol} and Dam Kim and Lee, {Hye Soon} and Criswell, {Lindsey A.} and Freedman, {Barry I.} and Gilkeson, {Gary S.} and Guthridge, {Joel M.} and Jacob, {Chaim O.} and James, {Judith A.} and Kamen, {Diane L.} and Merrill, {Joan T.} and Sivils, {Kathy Moser} and Niewold, {Timothy B.} and Michelle Petri and Rosalind Ramsey-Goldman and Reveille, {John D.} and Scofield, {R. Hal} and Stevens, {Anne M.} and Vil{\'a}, {Luis M.} and Vyse, {Timothy J.} and Kaufman, {Kenneth M.} and Harley, {John B.} and Langefeld, {Carl D.} and Gaffney, {Patrick M.} and Brown, {Elizabeth E.} and Edberg, {Jeffrey C.} and Kimberly, {Robert P.} and Daniela Ulgiati and Tsao, {Betty P.} and Boackle, {Susan A.} and Johan Frosteg{\aa}rd and Lennart Truedsson",
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T1 - Preferential association of a functional variant in complement receptor 2 with antibodies to double-stranded DNA

AU - BIOLUPUS and GENLES Networks

AU - Zhao, Jian

AU - Giles, Brendan M.

AU - Taylor, Rhonda L.

AU - Yette, Gabriel A.

AU - Lough, Kara M.

AU - Ng, Han Leng

AU - Abraham, Lawrence J.

AU - Wu, Hui

AU - Kelly, Jennifer A.

AU - Glenn, Stuart B.

AU - Adler, Adam J.

AU - Williams, Adrienne H.

AU - Comeau, Mary E.

AU - Ziegler, Julie T.

AU - Marion, Miranda

AU - Alarcón-Riquelme, Marta E.

AU - Alarcón, Graciela S.

AU - Anaya, Juan Manuel

AU - Bae, Sang Cheol

AU - Kim, Dam

AU - Lee, Hye Soon

AU - Criswell, Lindsey A.

AU - Freedman, Barry I.

AU - Gilkeson, Gary S.

AU - Guthridge, Joel M.

AU - Jacob, Chaim O.

AU - James, Judith A.

AU - Kamen, Diane L.

AU - Merrill, Joan T.

AU - Sivils, Kathy Moser

AU - Niewold, Timothy B.

AU - Petri, Michelle

AU - Ramsey-Goldman, Rosalind

AU - Reveille, John D.

AU - Scofield, R. Hal

AU - Stevens, Anne M.

AU - Vilá, Luis M.

AU - Vyse, Timothy J.

AU - Kaufman, Kenneth M.

AU - Harley, John B.

AU - Langefeld, Carl D.

AU - Gaffney, Patrick M.

AU - Brown, Elizabeth E.

AU - Edberg, Jeffrey C.

AU - Kimberly, Robert P.

AU - Ulgiati, Daniela

AU - Tsao, Betty P.

AU - Boackle, Susan A.

AU - Frostegård, Johan

AU - Truedsson, Lennart

PY - 2016/1/1

Y1 - 2016/1/1

N2 - Objectives Systemic lupus erythematosus (SLE; OMIM 152700) is characterised by the production of antibodies to nuclear antigens. We previously identified variants in complement receptor 2 (CR2/CD21) that were associated with decreased risk of SLE. This study aimed to identify the causal variant for this association. Methods Genotyped and imputed genetic variants spanning CR2 were assessed for association with SLE in 15 750 case-control subjects from four ancestral groups. Allele-specific functional effects of associated variants were determined using quantitative real-time PCR, quantitative flow cytometry, electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation (ChIP)-PCR. Results The strongest association signal was detected at rs1876453 in intron 1 of CR2 (pmeta=4.2×10-4, OR 0.85), specifically when subjects were stratified based on the presence of dsDNA autoantibodies (case-control pmeta=7.6×10-7, OR 0.71; case-only pmeta=1.9×10-4, OR 0.75). Although allele-specific effects on B cell CR2 mRNA or protein levels were not identified, levels of complement receptor 1 (CR1/CD35) mRNA and protein were significantly higher on B cells of subjects harbouring the minor allele (p=0.0248 and p=0.0006, respectively). The minor allele altered the formation of several DNA protein complexes by EMSA, including one containing CCCTC-binding factor (CTCF), an effect that was confirmed by ChIP-PCR. Conclusions These data suggest that rs1876453 in CR2 has long-range effects on gene regulation that decrease susceptibility to lupus. Since the minor allele at rs1876453 is preferentially associated with reduced risk of the highly specific dsDNA autoantibodies that are present in preclinical, active and severe lupus, understanding its mechanisms will have important therapeutic implications.

AB - Objectives Systemic lupus erythematosus (SLE; OMIM 152700) is characterised by the production of antibodies to nuclear antigens. We previously identified variants in complement receptor 2 (CR2/CD21) that were associated with decreased risk of SLE. This study aimed to identify the causal variant for this association. Methods Genotyped and imputed genetic variants spanning CR2 were assessed for association with SLE in 15 750 case-control subjects from four ancestral groups. Allele-specific functional effects of associated variants were determined using quantitative real-time PCR, quantitative flow cytometry, electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation (ChIP)-PCR. Results The strongest association signal was detected at rs1876453 in intron 1 of CR2 (pmeta=4.2×10-4, OR 0.85), specifically when subjects were stratified based on the presence of dsDNA autoantibodies (case-control pmeta=7.6×10-7, OR 0.71; case-only pmeta=1.9×10-4, OR 0.75). Although allele-specific effects on B cell CR2 mRNA or protein levels were not identified, levels of complement receptor 1 (CR1/CD35) mRNA and protein were significantly higher on B cells of subjects harbouring the minor allele (p=0.0248 and p=0.0006, respectively). The minor allele altered the formation of several DNA protein complexes by EMSA, including one containing CCCTC-binding factor (CTCF), an effect that was confirmed by ChIP-PCR. Conclusions These data suggest that rs1876453 in CR2 has long-range effects on gene regulation that decrease susceptibility to lupus. Since the minor allele at rs1876453 is preferentially associated with reduced risk of the highly specific dsDNA autoantibodies that are present in preclinical, active and severe lupus, understanding its mechanisms will have important therapeutic implications.

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