ABSTRACT
This study was undertaken to evaluate the possibility to obtain a molecular signature of rheumatoid arthritis (RA) comparatively osteoarthritis (OA), and to lay the bases to develop new diagnostic tools and identify new targets. Microarray technology was used for such an analysis. The gene expression profiles of synovial tissues from patients with confirmed RA, and patients with OA were established and compared. A set of 63 genes was selected, based, more specifically, on their overexpression or underexpression in RA samples compared to OA. Results for six of these genes have been verified by quantitative PCR using both samples identical to those used in the microarray experiments and entirely separate samples. Expression profile of the 48 known genes allowed the correct classification of additional RA and OA patients. Furthermore, the distinct expression of three of the selected genes was also studied by quantitative RT-PCR in cultured synovial cells. Detailed analysis of the expression profile of the selected genes provided evidence for dysregulated biological pathways, pointed out to chromosomal location and revealed novel genes potentially involved in RA. It is proposed that such an approach allows valuable diagnosis/prognostics tools in RA to be established and potential targets for combating the disease to be identified.
Subject(s)
Arthritis, Rheumatoid/genetics , Arthritis, Rheumatoid/physiopathology , Gene Expression , Oligonucleotide Array Sequence Analysis/methods , Osteoarthritis/genetics , RNA, Messenger/metabolism , Adult , Aged , Aged, 80 and over , Cathepsin L , Cathepsins/genetics , Cathepsins/metabolism , Cells, Cultured , Clusterin , Cysteine Endopeptidases , DEAD-box RNA Helicases , DNA Primers , Female , GTP-Binding Proteins/genetics , GTP-Binding Proteins/metabolism , Glycoproteins/genetics , Glycoproteins/metabolism , Humans , Lactoylglutathione Lyase/genetics , Lactoylglutathione Lyase/metabolism , Male , Membrane Proteins/genetics , Membrane Proteins/metabolism , Middle Aged , Molecular Chaperones/genetics , Molecular Chaperones/metabolism , Nucleic Acid Hybridization , Polymerase Chain Reaction , RNA Helicases/genetics , RNA Helicases/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Synovial Fluid/metabolismSubject(s)
Chromosomes, Human, Pair 13 , Connexins/genetics , Ectodermal Dysplasia/genetics , Mutation, Missense , Amino Acid Sequence , Amino Acid Substitution , Animals , Arginine , Chromosome Mapping , Connexin 30 , Female , Glycine , Humans , Male , Mice , Molecular Sequence Data , Pedigree , Sequence Alignment , Sequence Homology, Amino AcidABSTRACT
The HED (hidrotic ectodermal dysplasia) or Clouston syndrome gene (named ED2) has been mapped to the pericentromeric region of chromosome 13 (13q11) to a 2.4-cM interval flanked by markers D13S1828 and D13S1830. We have developed a BAC/PAC-based contig map of this region. This contig, comprising 23 clones and spanning 1.5 Mb, was established by mapping of 27 BAC/PAC end-derived STSs, 11 known polymorphic markers, 2 previously mapped genes, and 14 ESTs. The genomic clone overlaps were confirmed by restriction fragment fingerprint analysis. This contig provides the basis for genomic sequencing and gene identification in the ED2 critical region. Of the 14 ESTs mapped to the contig, 6 show homology to human genes and 8 appear to be novel. Expression patterns of the genes/ESTs were tested by Northern blot and RT-PCR. Full characterization of some of these genes, as well as the novel ESTs, will be useful in assessing their involvement in the HED/Clouston syndrome.
