Chromatin immunoprecipitation (ChIP)assays identify a functional retinoic acid receptorbinding site and−549T>C SNP in the PTGDR‐5′region

  1. García-Sánchez A 1
  2. Marcos-Vadillo E 2
  3. Sanz C 1
  4. Estravís M 2
  5. Cornejo-García JA 3
  6. Moreno-Rodilla E 4
  7. Dávila I 4
  8. Isidoro-García M 5
  1. 1 Universidad de Salamanca
    info

    Universidad de Salamanca

    Salamanca, España

    ROR https://ror.org/02f40zc51

  2. 2 Hospital Universitario de Salamanca
    info

    Hospital Universitario de Salamanca

    Salamanca, España

    ROR https://ror.org/0131vfw26

  3. 3 Hospital Regional Universitario de Málaga
    info

    Hospital Regional Universitario de Málaga

    Málaga, España

    ROR https://ror.org/01mqsmm97

  4. 4 IBSAL, Institute ofBiomedical Research of Salamanca; Department of Biomedical andDiagnostic Sciences. University of Salamanca; Department ofImmunoallergy. Salamanca University Hospital
  5. 5 IBSAL, Institute of Biomedical Research of Salamanca; Department of Clinical Biochemistry; Department of Medicine. University of Salamanca
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Actas:
Allergy

ISSN: 0105-4538,  1398-9995

Año de publicación: 2018

Volumen: 73

Número: s103

Páginas: 263-263

Congreso: European Academy of Allergy and Clinical Immunology Congress

Tipo: Aportación congreso

GOOGLE SCHOLAR

Resumen

Background:The Prostaglandin D2 receptor (PTGDR), participatesin airways inflammation in allergic asthma. Functional studies suggestthat promoter polymorphisms have a role in the susceptibility to suf-fer asthma. Retinoic acid (ATRA) has been linked to allergic diseases.We have described thePTGDRpromoter activation mediated byATRA through response elements (RARE) at position−549T>C. Weaimed to investigate the binding of the ATRA receptor (RAR) in thepredicted RARE sequences of the PTGDR promoter.Method:Chromatin immunoprecipitation assays (ChIP) were per-formed in the KU812 line, stimulated with 1μmol/L ATRA or DMSOand in peripheral blood mononuclear cells (PBMC) of patients carry-ing the−549C or−549T variants. Immunoprecipitated DNA withanti‐RARαand anti‐RARβantibodies was analyzed by qRT‐PCR.Results:We observed an enrichment of target DNA fragmentsimmunoprecipitated with both anti‐RARαandβantibodies in KU812cells treated with 1μmol/L ATRA. The stimulation by ATRA signifi-cantly promoted the binding of RAR to the RARE sequence withinthe−549T polymorphism. No RAR binding was detected when thedistal region of thePTGDRintron was amplified (P<0.001, Kruskal‐Wallis test). When we performed ChIP on PBMC of subjects carryingthe−549T polymorphism, we also observed a higher enrichmentwith anti‐RARβof the immunoprecipitated DNA in PBMC comparedto patients carrying the−549C variant.Conclusion:The presence of the−549T>C polymorphism influ-ences the binding of ATRA to regulatory elements of thePTGDRpromoter. Genetic differences in the regulation ofPTGDRby ATRAcould contribute to the phenotypic differences observed in allergicpatients.