title: CALU polymorphism A29809G affects calumenin availability involving vascular calcification.
abstract: Calumenin inhibits gamma-carboxylation of matrix-Gla-protein preventing BMP2-dependent calcification. Our aim was to explore the clinical relevance and functionality of the CALU polymorphism rs1043550, and the relationship of calumenin time-dependent expression profile with the active calcification of human vascular smooth muscle cells (hVSMC). Coronary artery calcium score and lesion severity were assessed by cardiac computed tomography in 139 consecutive low-risk patients genotyped for rs1043550. Polymorphic (G) allele carriage was associated with lower calcium (OR: 6.19, p=0.042). Calcified arteries from CALU 'A' allele carriers undergoing cardiovascular surgery exhibited higher residual calcification, higher calumenin immunostaining and lower matrix-Gla-protein, contrary to 'G' allele carriers. In a luciferase reporter system in vascular cells, polymorphic 'G' allele reduced the mRNA stability by 30% (p < 0.05). Osteogenic high-phosphate media induced active differentiation of hVSMC onto functional osteoblast-like cells as demonstrated by extracellular matrix mineralization and osteoblast markers expression. Calumenin was early over-expressed at day 3 (p < 0.05), but decreased thereafter (mRNA and protein) with implications on gamma-carboxylation system. Calumenin was found released and co-localizing with extracellular matrix calcifications. The CALU polymorphism rs1043550 affects mRNA stability and tissue availability of calumenin thus supporting the protective clinical significance. Calumenin shows a time-dependent profile during induced calcification. These data demonstrate a novel association of vascular calcification with the VSMC phenotypic transition into osteoblast-like cells. Moreover, hyperphosphatemic stimuli render calumenin accumulation in the mineralized extracellular matrix.
authors.affiliation: Hospital Clínico Universitario Virgen de la Arrixaca, Universidad de Murcia, IMIB-Arrixaca, Murcia, Spain.
authors.forename: Eva
authors.initials: E
authors.lastname: Jover
authors.affiliation: Hospital Clínico Universitario Virgen de la Arrixaca, Universidad de Murcia, IMIB-Arrixaca, Murcia, Spain. Electronic address: fcomarino@hotmail.com.
authors.forename: Francisco
authors.initials: F
authors.lastname: Marín
authors.affiliation: Hospital Clínico Universitario Virgen de la Arrixaca, Universidad de Murcia, IMIB-Arrixaca, Murcia, Spain.
authors.forename: Míriam
authors.initials: M
authors.lastname: Quintana
authors.affiliation: Hospital Clínico Universitario Virgen de la Arrixaca, Universidad de Murcia, IMIB-Arrixaca, Murcia, Spain.
authors.forename: Joaquín
authors.initials: J
authors.lastname: Pérez-Andreu
authors.affiliation: Hospital Clínico Universitario Virgen de la Arrixaca, Universidad de Murcia, IMIB-Arrixaca, Murcia, Spain.
authors.forename: José Antonio
authors.initials: JA
authors.lastname: Hurtado
authors.affiliation: Centro de Investigación Cardiovascular, CSIC-ICCC, Instituto de Investigación Biomédica Sant Pau (IIB Sant Pau), Barcelona, Spain.
authors.forename: Cristina
authors.initials: C
authors.lastname: Rodríguez
authors.affiliation: Centro de Investigación Cardiovascular, CSIC-ICCC, Instituto de Investigación Biomédica Sant Pau (IIB Sant Pau), Barcelona, Spain.
authors.forename: José
authors.initials: J
authors.lastname: Martínez-González
authors.affiliation: Centro Regional de Hemodonación, Murcia, Spain.
authors.forename: Rocío
authors.initials: R
authors.lastname: González-Conejero
authors.affiliation: Hospital Clínico Universitario Virgen de la Arrixaca, Universidad de Murcia, IMIB-Arrixaca, Murcia, Spain.
authors.forename: Mariano
authors.initials: M
authors.lastname: Valdés
authors.affiliation: Hospital Clínico Universitario Virgen de la Arrixaca, Universidad de Murcia, IMIB-Arrixaca, Murcia, Spain.
authors.forename: Diana
authors.initials: D
authors.lastname: Hernández-Romero
mesh_terms: D000483:Alleles; D002118:Calcium / Q000378:metabolism; D002135:Calcium-Binding Proteins / Q000235:genetics* / Q000378:metabolism*; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002478:Cells, Cultured; D003331:Coronary Vessels / Q000378:metabolism / Q000473:pathology; D005109:Extracellular Matrix / Q000378:metabolism; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D006801:Humans; D009131:Muscle, Smooth, Vascular / Q000378:metabolism / Q000473:pathology; D032389:Myocytes, Smooth Muscle / Q000166:cytology / Q000378:metabolism; D010006:Osteoblasts / Q000166:cytology / Q000378:metabolism; D020641:Polymorphism, Single Nucleotide; D020871:RNA Stability / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D061205:Vascular Calcification / Q000235:genetics* / Q000378:metabolism*; D000097568:Matrix Gla Protein
chemical_list: D002118:Calcium; D002135:Calcium-Binding Proteins; D016326:Extracellular Matrix Proteins; D012333:RNA, Messenger; D000097568:Matrix Gla Protein; C510527:CALU protein, human
keywords: Calumenin; Gamma-carboxylation; Vascular calcification; Vascular smooth muscle cell; rs1043550