title: Vitamin D receptor gene (VDR) transcripts in bone, cartilage, muscles and blood and microarray analysis of vitamin D responsive genes expression in paravertebral muscles of juvenile and adolescent idiopathic scoliosis patients.
abstract: BACKGROUND
  VDR may be considered as a candidate gene potentially related to idiopathic scoliosis susceptibility and natural history. Transcriptional profile of VDR mRNA isoforms might be changed in the structural tissues of the scoliotic spine and potentially influence the expression of VDR responsive genes. The purpose of the study was to determine differences in mRNA abundance of VDR isoforms in bone, cartilage and paravertebral muscles between tissues from curve concavity and convexity, between JIS and AIS and to identify VDR responsive genes differentiating juvenile and adolescent idiopathic scoliosis in paravertebral muscles.
  
  
  METHODS
  In a group of 29 patients with JIS and AIS, specimens of bone, cartilage, paravertebral muscles were harvested at the both sides of the curve apex together with peripheral blood samples. Extracted total RNA served as a matrix for VDRs and VDRl mRNA quantification by QRT PCR. Subsequent microarray analysis of paravertebral muscular tissue samples was performed with HG U133A chips (Affymetrix). Quantitative data were compared by a nonparametric Mann Whitney U test. Microarray results were analyzed with GeneSpring 11GX application. Matrix plot of normalized log-intensities visualized the degree of differentiation between muscular tissue transcriptomes of JIS and AIS group. Fold Change Analysis with cutoff of Fold Change ≥2 identified differentially expressed VDR responsive genes in paravertebral muscles of JIS and AIS.
  
  
  RESULTS
  No significant differences in transcript abundance of VDR isoforms between tissues of the curve concavity and convexity were found. Statistically significant difference between JIS and AIS group in mRNA abundance of VDRl isoform was found in paravertebral muscles of curve concavity. Higher degree of muscular transcriptome differentiation between curve concavity and convexity was visualized in JIS group. In paravertebral muscles Tob2 and MED13 were selected as genes differentially expressed in JIS and AIS group.
  
  
  CONCLUSIONS
  In Idiopathic Scolioses transcriptional activity and alternative splicing of VDR mRNA in osseous, cartilaginous, and paravertebral muscular tissues are tissue specific and equal on both sides of the curve. The number of mRNA copies of VDRl izoform in concave paravertebral muscles might be one of the factors differentiating JIS and AIS. In paravertebral muscles Tob2 and Med13 genes differentiate Adolescent and Juvenile type of Idiopathic Scoliosis.
authors.affiliation: Orthopaedics Clinic Medical University of Silesia, Wojewódzki Szpital Specjalistyczny nr5 41-200 Sosnowiec, Pl. Medyków 1, Poland. nowrom20@gmail.com
authors.forename: Roman
authors.initials: R
authors.lastname: Nowak
authors.forename: Justyna
authors.initials: J
authors.lastname: Szota
authors.forename: Urszula
authors.initials: U
authors.lastname: Mazurek
mesh_terms: D000293:Adolescent; D017668:Age of Onset; D001842:Bone and Bones / Q000737:chemistry*; D002356:Cartilage / Q000737:chemistry*; D018797:Cell Cycle Proteins / Q000235:genetics; D002648:Child; D002675:Child, Preschool; D018450:Disease Progression; D005260:Female; D020869:Gene Expression Profiling / Q000379:methods*; D005786:Gene Expression Regulation; D005819:Genetic Markers; D020022:Genetic Predisposition to Disease; D006801:Humans; D008297:Male; D056892:Mediator Complex / Q000235:genetics; D018482:Muscle, Skeletal / Q000737:chemistry*; D020411:Oligonucleotide Array Sequence Analysis; D010641:Phenotype; D011044:Poland / Q000453:epidemiology; D012333:RNA, Messenger / Q000097:blood*; D060888:Real-Time Polymerase Chain Reaction; D018167:Receptors, Calcitriol / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012600:Scoliosis / Q000097:blood / Q000145:classification / Q000000981:diagnostic imaging / Q000453:epidemiology / Q000235:genetics*; D012720:Severity of Illness Index; D013131:Spine / Q000000981:diagnostic imaging; D014057:Tomography, X-Ray Computed
chemical_list: D018797:Cell Cycle Proteins; D005819:Genetic Markers; C488666:MED13 protein, human; D056892:Mediator Complex; D012333:RNA, Messenger; D018167:Receptors, Calcitriol; C402648:TOB2 protein, human; C579457:VDR protein, human