Abstract
Familial hyperkalemic hypertension (FHHt) is a Mendelian form of arterial hypertension that is partially explained by mutations in WNK1 and WNK4 that lead to increased activity of the Na+-Cl− cotransporter (NCC) in the distal nephron. Using combined linkage analysis and whole-exome sequencing in two families, we identified KLHL3 as a third gene responsible for FHHt. Direct sequencing of 43 other affected individuals revealed 11 additional missense mutations that were associated with heterogeneous phenotypes and diverse modes of inheritance. Polymorphisms at KLHL3 were not associated with blood pressure. The KLHL3 protein belongs to the BTB-BACK-kelch family of actin-binding proteins that recruit substrates for Cullin3-based ubiquitin ligase complexes. KLHL3 is coexpressed with NCC and downregulates NCC expression at the cell surface. Our study establishes a role for KLHL3 as a new member of the complex signaling pathway regulating ion homeostasis in the distal nephron and indirectly blood pressure.
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Change history
22 March 2012
In the version of this article initially published, two references were omitted, resulting in several statements being incorrectly attributed in the Online Methods. In the 'Ped01' subsection of the 'Whole-exome sequencing' section, two statements were attributed to ref. 21. The correct reference for these statements has been added as ref. 27. In the 'Ped02' subsection of the 'Whole-exome sequencing' section, Annovar was incorrectly attributed to ref. 22. The correct reference has been added as ref. 28. As a result of the addition of these two references, former refs. 27-36 have been renumbered as refs. 29-38, respectively, in the text and reference list. In addition, author affiliation 7 was incorrectly given as Genes and Disease Program, Center for Genomic Regulation (CGR), Pompeu Fabra University, Barcelona, Spain. The correct affiliation is Genes and Disease Program, Center for Genomic Regulation (CGR) and Pompeu Fabra University, Barcelona, Spain. Similarly, affiliation 32 was incorrectly given as Genomic and Epigenetic Variation in Disease Group, Center for Genomic Regulation, Universitat Pompeu Fabra, Barcelona, Spain. The correct affiliation is Genomic and Epigenetic Variation in Disease Group, CGR and Pompeu Fabra University, Barcelona, Spain. These corrections have been made in the HTML and PDF versions of the article.
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Acknowledgements
We thank C. Büsst for her critical reading of the manuscript, E. Clauser, F. Auradé and C. Auzan for helpful discussions, E. Durand, S. Lecointe and F. De Graeve for their contribution to whole-exome sequencing and M. Longépée-Dupas for assistance in family screening. This work was supported by INSERM, the Agence Nationale pour la Recherche (ANR; 05-MRAR-010-01), the European Union Framework Programme 7 through the HYPERGENE project (HEALTH-F4-2007-201550) and through the GEUVADIS project (HEALTH-261123), the Leducq Foundation (Transatlantic Network on Hypertension; 07 CVD 01), the Fondation Leducq Trans-Atlantic Network of Excellence (05 CVD 01; Preventing Sudden Death), the Groupe de Réflexion sur la Recherche Cardiovasculaire and Biotronik and the Ministry of Science and Innovation (MICINN; SAF2008-00357).
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H.L.-D.-P. designed, performed and analyzed the data of the genetic analyses and immunoblotting experiments, and wrote the corresponding parts of the manuscript. J.B. designed, performed and analyzed the data of the genetic analyses (Ped02). D.T. together with S.O. and X.E. designed, performed and analyzed the WES of the Ped02 kindred. S.M.-L. designed and performed the in vitro and immunofluorescence experiments, and wrote the corresponding parts of the manuscript. N.B.-N. designed the WES for the Ped01 kindred and participated in the interpretation of the genetic analyses (exome sequencing of Ped01 and association studies) and in the writing of the manuscript. O.P. designed, performed and analyzed KLHL3 modeling and the prediction of the consequences of KLHL3 mutations. G.B. participated in the collection of the families with FHHt and to the analysis of previous candidate genes. V.E. collected and characterized the first members of the Nantes FHHt family. A.B. contributed to the WES of Ped01. O.S. performed the bioinformatics analyses of WES in Ped01. C. Simian performed the sequencing of hypertensive and normotensive individuals. E.V.-P. designed, performed and analyzed the qRT-PCR experiments. C. Soukaseum provided technical help to H.L.-D.-P. C. Mandet performed the immunohistochemistry. F.B., O.C., M.D., B.F., P.H., J.K., M.K., P.L., C. Mourani, P.N., V.P., C.T., R.J.U., S.D.S. and C.I.B. recruited families with FHHt. G.E. and M.C. designed, performed and analyzed the association study of KLHL3 and blood pressure within the ICBP. P.B. designed and analyzed the immunohistochemistry experiments. P.F. designed and analyzed WES. J.-J.S. designed and analyzed the genetic study and wrote the corresponding parts of the manuscript. X.J. and J.H. designed the study, analyzed data and co-wrote the manuscript. X.J. organized the entire study.
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Supplementary Figures 1–6, Supplementary Tables 1–3, 5 and 6 and Supplementary Note (PDF 21357 kb)
Supplementary Table 4
Association of common variants at the KLHL3 locus with blood pressure in healthy individuals (XLS 255 kb)
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Louis-Dit-Picard, H., Barc, J., Trujillano, D. et al. KLHL3 mutations cause familial hyperkalemic hypertension by impairing ion transport in the distal nephron. Nat Genet 44, 456–460 (2012). https://doi.org/10.1038/ng.2218
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DOI: https://doi.org/10.1038/ng.2218
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