Analyses of RANK and RANKL in the Post-GWAS Context: Functional Evidence of Vitamin D Stimulation Through a RANKL Distal Region

Authors

  • Guy Yoskovitz,

    1. URFOA, IMIM (Institut Hospital del Mar d'Investigacions Mèdiques), Red Temática de Investigación Cooperativa en Envejecimiento y Fragilidad (RETICEF), Barcelona, Spain
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  • Natalia Garcia-Giralt,

    1. URFOA, IMIM (Institut Hospital del Mar d'Investigacions Mèdiques), Red Temática de Investigación Cooperativa en Envejecimiento y Fragilidad (RETICEF), Barcelona, Spain
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  • Maria Rodriguez-Sanz,

    1. URFOA, IMIM (Institut Hospital del Mar d'Investigacions Mèdiques), Red Temática de Investigación Cooperativa en Envejecimiento y Fragilidad (RETICEF), Barcelona, Spain
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  • Roser Urreizti,

    1. Departament de Genètica, Universitat de Barcelona, IBUB, Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER), ISCIII, Barcelona, Spain
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  • Robert Guerri,

    1. URFOA, IMIM (Institut Hospital del Mar d'Investigacions Mèdiques), Red Temática de Investigación Cooperativa en Envejecimiento y Fragilidad (RETICEF), Barcelona, Spain
    2. Servei de Medicina Interna, Hospital del Mar, Universitat Autònoma de Barcelona (UAB), Barcelona, Spain
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  • Sergi Ariño-Ballester,

    1. URFOA, IMIM (Institut Hospital del Mar d'Investigacions Mèdiques), Red Temática de Investigación Cooperativa en Envejecimiento y Fragilidad (RETICEF), Barcelona, Spain
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  • Daniel Prieto-Alhambra,

    1. URFOA, IMIM (Institut Hospital del Mar d'Investigacions Mèdiques), Red Temática de Investigación Cooperativa en Envejecimiento y Fragilidad (RETICEF), Barcelona, Spain
    2. University of Oxford, NIHR Biomedical Research Unit, IDIAP Jordi Gol, Institut Català de la Salut, Primary Care, Barcelona, Spain
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  • Leonardo Mellibovsky,

    1. URFOA, IMIM (Institut Hospital del Mar d'Investigacions Mèdiques), Red Temática de Investigación Cooperativa en Envejecimiento y Fragilidad (RETICEF), Barcelona, Spain
    2. Servei de Medicina Interna, Hospital del Mar, Universitat Autònoma de Barcelona (UAB), Barcelona, Spain
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  • Daniel Grinberg,

    1. Departament de Genètica, Universitat de Barcelona, IBUB, Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER), ISCIII, Barcelona, Spain
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  • Xavier Nogues,

    1. URFOA, IMIM (Institut Hospital del Mar d'Investigacions Mèdiques), Red Temática de Investigación Cooperativa en Envejecimiento y Fragilidad (RETICEF), Barcelona, Spain
    2. Servei de Medicina Interna, Hospital del Mar, Universitat Autònoma de Barcelona (UAB), Barcelona, Spain
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  • Susana Balcells,

    1. Departament de Genètica, Universitat de Barcelona, IBUB, Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER), ISCIII, Barcelona, Spain
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  • Adolfo Diez-Perez

    Corresponding author
    1. URFOA, IMIM (Institut Hospital del Mar d'Investigacions Mèdiques), Red Temática de Investigación Cooperativa en Envejecimiento y Fragilidad (RETICEF), Barcelona, Spain
    2. Servei de Medicina Interna, Hospital del Mar, Universitat Autònoma de Barcelona (UAB), Barcelona, Spain
    • Address correspondence to: Adolfo Diez-Perez, MD, PhD, URFOA, IMIM (Institut Hospital del Mar d'Investigacions Mèdiques), Doctor Aiguader,88 Suite 200.10 08003 Barcelona, Spain, Red Temática de Investigación Cooperativa en Envejecimiento y Fragilidad (RETICEF), Barcelona, Spain. E-mail: ADiez@parcdesalutmar.cat

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ABSTRACT

Over the past decade, many genome-wide association studies (GWAs) and meta-analyses have identified genes and regions involved in osteoporotic phenotypes. Nevertheless, the large majority of these results were not tested at any functional level. GWA-associated single-nucleotide polymorphisms (SNPs) near candidate genes such as RANK and RANKL suggest that these SNPs and/or other variants nearby may be involved in bone phenotype determination. This study focuses on SNPs along these two genes, which encode proteins with a well-established role in the bone remodeling equilibrium. Thirty-three SNPs, chosen for their location in evolutionary conserved regions or replicated from previous studies, were genotyped in the BARCOS cohort of 1061 postmenopausal women and tested for association with osteoporotic phenotypes. SNP rs9594738, which lies 184 kb upstream of the RANKL gene, was the only SNP found to be associated with a bone phenotype (dominant model: beta coefficient = –0.034, p = 1.5 × 10−4, for lumbar spine bone mineral density). Functional experiments exploring a distal region (DR) of 831 bp that harbors this SNP in a centered position (nt 470) demonstrated its capacity to inhibit the RANKL promoter in reporter gene assays. Remarkably, this DR inhibition was significantly reduced in the presence of vitamin D. In conclusion, the GWA-associated SNP rs9594738 lies in a region involved in transcription regulation through which vitamin D could be regulating RANKL expression and bone mineral density. © 2013 American Society for Bone and Mineral Research.

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