Abstract

Pulmonary arterial hypertension (PAH) is a rare, severe disease resulting from progressive obliteration of small-caliber pulmonary arteries by proliferating vascular cells. PAH can occur without recognized etiology (idiopathic PAH), be associated with a systemic disease or occur as a heritable form, with BMPR2 mutated in approximately 80% of familial and 15% of idiopathic PAH cases1,2,3. We conducted a genome-wide association study (GWAS) based on 2 independent case-control studies for idiopathic and familial PAH (without BMPR2 mutations), including a total of 625 cases and 1,525 healthy individuals. We detected a significant association at the CBLN2 locus mapping to 18q22.3, with the risk allele conferring an odds ratio for PAH of 1.97 (1.59–2.45; P = 7.47 × 10−10). CBLN2 is expressed in the lung, and its expression is higher in explanted lungs from individuals with PAH and in endothelial cells cultured from explanted PAH lungs.

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Acknowledgements

We thank E. Fadel and O. Mercier from the Centre Médico-Chirurgical Marie-Lannelongue for their help in lung sample collection. The Pulmonary Hypertension Allele-Associated Risk (PHAAR) project was financially supported by the Agence Nationale pour la Recherche (Project ANR-07-MRAR-021) and by PHRC AOM07-041, INSERM and UPMC. The 3C Study is conducted under a partnership agreement between INSERM, the Victor Segalen–Bordeaux II University and Sanofi-Synthelabo. The Fondation pour la Recherche Médicale funded the preparation and first phase of the study. The 3C Study is also supported by the Caisse Nationale Maladie des Travailleurs Salariés, the Direction Générale de la Santé, the Mutuelle Générale de l'Education Nationale, the Institut de la Longévité, the Agence Française de Sécurité Sanitaire des Produits de Santé, the regional governments of Aquitaine, Bourgogne and Languedoc-Roussillon and the Fondation de France, and the Ministry of Research–INSERM Programme Cohorts and Collection of Biological Material. The Lille Genopole received an unconditional grant from Eisai. The financial supporters had no role in study design, data collection and analysis, decision to publish or preparation of the manuscript. M.G. was funded by a grant from the Agence Nationale pour la Recherche (Project PHAAR, ANR-07-MRAR-021) and the Program Hospitalier de Recherche Clinique (PHRC2009 RENOVA-TV). Statistical analyses used the C2BIG computing centre funded by the Fondation pour la Recherche Médicale and Région Ile de France. Collection and management of samples from Vanderbilt University were supported by US NIH grants P01 HL072058 and K23 HL0987431 and Vanderbilt General Clinical Research Center (GCRC) RR000095. Collection of the samples from Columbia University was supported by US NIH grant R01 HL060056.

Author information

Affiliations

  1. Unité Mixte de Recherche en Santé (UMRS) 937, Université Pierre & Marie Curie (UPMC) Université Paris 6 and Institut National de la Santé et de la Recherche Médicale (INSERM), Paris, France.

    • Marine Germain
    •  & David A Trégouët
  2. Institut Hospitalo-Universitaire (IHU) Cardiométabolisme et Nutrition (ICAN), Paris, France.

    • Marine Germain
    • , Mélanie Eyries
    • , Odette Poirier
    • , Sophie Nadaud
    • , Svetlana Maugenre
    • , David A Trégouët
    •  & Florent Soubrier
  3. UMRS 956, UPMC Université Paris 6 and INSERM, Paris, France.

    • Mélanie Eyries
    • , Odette Poirier
    • , Sophie Nadaud
    • , Svetlana Maugenre
    •  & Florent Soubrier
  4. Department of Genetics, Hôpital Pitié-Salpêtrière, Assistance Publique–Hôpitaux de Paris (AP-HP), Paris, France.

    • Mélanie Eyries
    • , Florence Coulet
    •  & Florent Soubrier
  5. Université Paris–Sud, Faculté de Médecine, Le Kremlin Bicêtre, France.

    • David Montani
    • , Barbara Girerd
    • , Peter Dorfmüller
    • , Christophe Guignabert
    • , Gerald Simonneau
    •  & Marc Humbert
  6. Centre National de Référence de l'Hypertension Pulmonaire Sévère, Département Hospitalo-Universitaire (DHU) Thorax Innovation, Service de Pneumologie, Hôpital de Bicêtre, AP-HP, Le Kremlin Bicêtre, France.

    • David Montani
    • , Barbara Girerd
    • , Peter Dorfmüller
    • , Christophe Guignabert
    • , Gerald Simonneau
    •  & Marc Humbert
  7. UMRS 999, INSERM and Université Paris–Sud, Laboratoire d'Excellence (LabEx) en Recherche sur le Médicament et l'Innovation Thérapeutique (LERMIT), Centre Chirurgical Marie Lannelongue, Le Plessis Robinson, France.

    • David Montani
    • , Barbara Girerd
    • , Peter Dorfmüller
    • , Christophe Guignabert
    • , Gerald Simonneau
    •  & Marc Humbert
  8. Department of Pathology, Centre Chirurgical Marie-Lannelongue, Le Plessis-Robinson, France.

    • Peter Dorfmüller
  9. Post-Genomic Platform (P3S), UPMC Université Paris 6, Paris, France.

    • Wassila Carpentier
  10. Department of Pulmonary Diseases, VU University Medical Center, Amsterdam, The Netherlands.

    • Anton Vonk-Noordegraaf
  11. Cardiac Surgery Department, Hôpital Necker–Enfants Malades, AP-HP, Paris, France.

    • Marilyne Lévy
  12. UMRS 765, INSERM and Université Paris Descartes, Paris, France.

    • Marilyne Lévy
  13. Respiratory Disease Department, Centre Hospitalier Universitaire (CHU) Brabois, Vandoeuvre-lès-Nancy, France.

    • Ari Chaouat
  14. UMRS 744, Université de Lille Nord de France and INSERM, Institut Pasteur, Lille, France.

    • Jean-Charles Lambert
    •  & Philippe Amouyel
  15. UMRS 708, UPMC Université Paris 6 and INSERM, Paris, France.

    • Marion Bertrand
  16. UMRS 888, Université Montpellier and INSERM, Montpellier, France.

    • Anne-Marie Dupuy
  17. U897, Université Bordeaux and INSERM, Institut de Santé Publique d'Epidémiologie et de Développement, Bordeaux, France.

    • Luc Letenneur
  18. Commissariat à l'Energie Atomique, Institut de Génomique, Centre National de Génotypage, Evry, France.

    • Mark Lathrop
  19. Centre Hospitalier Régional Universitaire de Lille, Lille, France.

    • Philippe Amouyel
  20. Centre for Human Genetics, University Hospitals of Leuven, Leuven, Belgium.

    • Thomy J L de Ravel
  21. Department of Pneumology, Catholic University of Leuven (KU Leuven), Leuven, Belgium.

    • Marion Delcroix
  22. Department of Pneumology, Gasthuisberg University Hospital, Leuven, Belgium.

    • Marion Delcroix
  23. Department of Pediatrics, Division of Pulmonary, Allergy, and Immunology Medicine, Vanderbilt University Medical Center, Nashville, Tennessee, USA.

    • Eric D Austin
  24. Department of Medicine, Division of Allergy, Pulmonary, and Critical Care Medicine, Vanderbilt University Medical Center, Nashville, Tennessee, USA.

    • Ivan M Robbins
    • , Anna R Hemnes
    •  & James E Loyd
  25. Department of Pediatrics, Columbia University, New York, New York, USA.

    • Erika Berman-Rosenzweig
    • , Robyn J Barst
    •  & Wendy K Chung

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Contributions

F.S. initiated and supervised the study. F.S., D.A.T., G.S. and M.H. conceived and designed the experiments. D.M., B.G., G.S., M.H., A.V.-N., M.Lévy, A.C., T.J.L.d.R. and M.D. analyzed clinical data of collected individuals with PAH for the discovery cohort. J.-C.L., M.B., A.-M.D., L.L., M.Lathrop and P.A. provided genotyped data for control subjects. E.D.A., I.M.R., A.R.H., J.E.L., E.B.-R., R.J.B. and W.K.C. analyzed the clinical data of the collected individuals with PAH for the replication cohort. M.E. and F.C. managed DNA samples. W.C. performed genotyping. M.G. and D.A.T. performed statistical analysis. M.E., O.P., S.N. and S.M. performed functional analyses. C.G. isolated vascular cells. P.D. performed tissue imaging. D.A.T., M.G., M.E., O.P., S.N. and F.S. analyzed data. F.S., M.G., M.E. and D.A.T. wrote the manuscript. O.P., S.N., M.H., D.M., E.D.A., J.E.L. and W.K.C. reviewed the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Florent Soubrier.

Supplementary information

PDF files

  1. 1.

    Supplementary Text and Figures

    Supplementary Figures 1–6 and Supplementary Tables 1, 3, 5 and 6

Excel files

  1. 1.

    Supplementary Table 2

    Association in the replication cohort of the 319 most significant SNPs with i/fPAH detected in the discovery GWAS

  2. 2.

    Supplementary Table 4

    Association analysis of all SNPs located within a 100kb upstream or downstream distance from the CBLN2 gene with ("conditional") and without adjusting for the rs2217560 effect in the discovery GWAS sample.

About this article

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DOI

https://doi.org/10.1038/ng.2581

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