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DNA Research 2006 13(4):169-183; doi:10.1093/dnares/dsl006
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© The Author 2006. Kazusa DNA Research Institute
The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact journals.permissions@oxfordjournals.org

Isolation and Expression Profiling of Genes Upregulated in Bone Marrow-Derived Mononuclear Cells of Rheumatoid Arthritis Patients

Nobuo Nakamura1 {dagger}, Yasunori Shimaoka2 {dagger}, Takahiro Tougan3, Hiroaki Onda3,4, Daisuke Okuzaki3, Hanjun Zhao3, Azumi Fujimori3, Norikazu Yabuta3, Ippei Nagamori3, Akie Tanigawa4, Jun Sato3, Takenori Oda5, Kenji Hayashida6, Ryuji Suzuki7, Masao Yukioka2, Hiroshi Nojima3,4,* and Takahiro Ochi7

1 Center of Arthroplasty, Kyowakai Hospital Suita, Japan
2 Yukioka Hospital Osaka, Japan
3 Department of Molecular Genetics, Research Institute for Microbial Diseases, Osaka University 3-1 Yamadaoka, Suita, Osaka 562-0031, Japan
4 Innovation Plaza Osaka Izumi, Japan
5 Department of Rheumatology, NHO Osaka-Minami Medical Center Kawachinagano, Japan
6 Hoshigaoka Kosei-Nenkin Hospital Hirakata, Japan
7 Clinical Research Center for Allergy and Rheumatology, National Sagamihara Hospital 18-1 Sakura-dai, Sagamihara, Kanagawa 228-8522, Japan

We have comprehensively identified the genes whose expressions are augmented in bone marrow-derived mononuclear cells (BMMC) from patients with Rheumatoid Arthritis (RA) as compared with BMMCs from Osteoarthritis (OA) patients, and named them AURA after augmented in RA. Both stepwise subtractive hybridization and microarray analyses were used to identify AURA genes, which were confirmed by northern blot analysis and/or reverse transcription polymerase chain reaction (RT–PCR). We also assessed their expression levels in individual patients by quantitative real-time RT–PCR. Of 103 AURA genes we have identified, the mRNA levels of the following 10 genes, which are somehow related to immune responses, were increased in many of the RA patients: AREG (=AURA9), FK506-binding protein 5 (FKBP5 = AURA45), C-type lectin superfamily member 9 (CLECSF9 = AURA24), tyrosylprotein sulfotransferase 1 (TPST1 = AURA52), lymphocyte G0/G1 switch gene (G0S2 = AURA8), chemokine receptor 4 (CXCR4 = AURA86), nuclear factor-kappa B (NF-{kappa}B = AURA25) and two genes of unknown function (FLJ11106 = AURA1, BC022398 [GenBank] = AURA2 and XM_058513 = AURA17). Since AREG was most significantly increased in many of the RA patients, we subjected it to further analysis and found that AREG-epidermal growth factor receptor signaling is highly activated in synovial cells isolated from RA patients, but not in OA synoviocytes. We propose that the expression profiling of these AURA genes may improve our understanding of the pathogenesis of RA.

Key words: stepwise subtraction; microarray; RA; OA; amphiregulin; synoviolin

*To whom correspondence should be addressed. Tel. +81-6-6875-3980, Fax. +81-6-6875-5192, E-mail: snj-0212{at}biken.osaka-u.ac.jp

Communicated by Mitsuo Oshimura

{dagger}These authors contributed equally to this work.


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