変形性関節症の新規治療法開発を目指したNotchシグナルの網羅的解析

2011 Fiscal Year Annual Research Report

• Project/Area Number: 23390358
• Research Institution: The University of Tokyo
• Principal Investigator: 筑田 博隆 東京大学, 医学部附属病院, 特任講師 (30345219)
• Co-Investigator(Kenkyū-buntansha): 川口 浩 東京大学, 医学部附属病院, 准教授 (40282660) ; 大橋 暁 東京大学, 医学部附属病院, 助教 (20466767) ; 唐司 寿一 東京大学, 医学部附属病院, 助教 (40597762)
• Keywords: 関節軟骨 / 変形性関節症 / Notchシグナル

Research Abstract

Notchシグナル関連分子として、受容体Notch1,2,3,4,リガンドDelta-like1,3,4,Jagged1,2、切断酵素ADAM10,17、下流のHes,Heyファミリーの発現を正常関節軟骨、変性関節軟骨において免疫組織染色にて調べたところ、Notch1,2の発現が強く、関節軟骨の変性に従って核内に移行していることが判明した。リガンドではJagged1の発現が関節軟骨の変性に従って増強しており、下流ではHes1が唯一強く発現が検出された。ADAM10,17の発現はともに強く検出された。次にNotchシグナルの核内共役分子Rbpjを軟骨特異的かつ薬剤誘導性にノックアウトして変形性関節症モデルを作成したところ、軟骨変性が有意に抑制された。組織切片において免疫組織染色を行ったところ、MMP13などの軟骨変性を惹起する分子がRbpjのノックアウトによって発現低下していることが分かった。次に軟骨系細胞株ATDC5にレトロウイルスを用いてNotch1細胞内ドメインを強制発現させたところ、Hes1のほか、MMP13、VEGFなどの分子が強く誘導されることが明らかとなった。またこれらの軟骨変性分子の誘導は、siRNAによるHes1の発現抑制を行うことによってキャンセルされることも明らかとなった。
またNotchシグナルの阻害剤であるDAPTを培養関節軟骨に投与して変性負荷をかけたところ、変性は有意に抑制され、Hes1のほか、MMP13、VEGFなどの発現も有意に抑制された。

Current Status of Research Progress

1: Research has progressed more than it was originally planned.

Reason

薬剤誘導性のコンディショナルノックアウトマウスを用いた解析で明らかな差が認められ、その前後で活性化している分子群を初年度に同定することができた。

Strategy for Future Research Activity

コンディショナルトランスジェニックマウスを用いた解析や、さらなるNotchシグナルの標的分子の探索を継続していく。

Research Products

• [Journal Article] C/EBPβ and RUNX2 cooperate to degrade cartilage with MMP-13 as the target and HIF-2α as the inducer in chondrocytes (2012)
  • Author(s): Hirata M, Kugimiya F, Fukai A, Saito T, Yano F, Ikeda T, Mabuchi A, Sapkota BR, Akune T, Nishida N, Yoshimura N, Nakagawa T, Tokunaga K, Nakamura K, Chung UI, Kawaguchi H
  • Journal Title: Hum Mol Genet Volume: 21 Pages: 1111-23
• [Journal Article] Lack of a chondroprotective effect of cyclooxygenase 2 inhibition in a surgically induced model of osteoarthritis in mice (2012)
  • Author(s): Fukai A, Kamekura S, Chikazu D, Nakagawa T, Hirata M, Saito T, Hosaka Y, Ikeda T, Nakamura K, Chung UI, Kawaguchi H
  • Journal Title: Arthritis Rheum Volume: 64 Pages: 198-203
• [Journal Article] Incidence and risk factors for radiographic knee osteoarthritis and knee pain in Japanese men and women : A longitudinal population-based cohort study (2012)
  • Author(s): Muraki S, Kawaguchi H, et al.
  • Journal Title: Arthritis Rheum Volume: (E-pub)
  • Peer Reviewed
• [Journal Article] Mortality and morbidity in dialysis-dependent patients undergoing spinal surgery : analysis of a national administrative database in Japan (2012)
  • Author(s): Chikuda H, et al.
  • Journal Title: J Bone Joint Surg Am Volume: 94 Pages: 433-8
  • DOI: PMID:22398737
  • Peer Reviewed
• [Journal Article] The normative score and the cut-off value of the Oswestry Disability Index (ODI) (2012)
  • Author(s): Tonosu J, Chikuda H, et al.
  • Journal Title: Eur Spine J Volume: (E-pub)
  • DOI: PMID:22298236
  • Peer Reviewed
• [Journal Article] Reference values for hand grip strength, muscle mass, walking time, and one-leg standing time as indices for locomotive syndrome and associated disability : the second survey of the ROAD study (2011)
  • Author(s): Yoshimura N, Kawaguchi H, et al.
  • Journal Title: J Orthop Sci Volume: 16 Pages: 768-77
  • Peer Reviewed
• [Journal Article] Independent association of joint space narrowing and osteophyte formation at the knee with health-related quality of life in Japan : a cross-sectional study (2011)
  • Author(s): Muraki S, Kawaguchi H, et al.
  • Journal Title: Arthritis Rheum Volume: 63 Pages: 3859-64
  • Peer Reviewed
• [Journal Article] Prevalence of falls and the association with knee osteoarthritis and lumbar spondylosis as well as knee and lower back pain in Japanese men and women (2011)
  • Author(s): Muraki S, Kawaguchi H, et al.
  • Journal Title: Arthritis Care Res Volume: 63 Pages: 1425-31
  • Peer Reviewed
• [Journal Article] Changes in serum levels of biochemical markers of bone turnover during 10 years among Japanese men and women : associated factors and birth-cohort effect. The Taiji Study (2011)
  • Author(s): Yoshimura N, Kawaguchi H, et al.
  • Journal Title: J Bone Miner Metab. Volume: 29 Pages: 699-708
  • Peer Reviewed
• [Journal Article] Health-related quality of life in subjects with low back pain and knee pain in a population-based cohort study of Japanese men : the Research on Osteoarthritis Against Disability study (2011)
  • Author(s): Muraki S, Kawaguchi H, et al.
  • Journal Title: Spine Volume: 36 Pages: 1312-9
  • Peer Reviewed
• [Journal Article] Biochemical markers of bone turnover as predictors of osteoporosis and osteoporotic fractures in men and women : 10-year follow-up of the Taiji cohort (2011)
  • Author(s): Yoshimura N, Kawaguchi H, et al.
  • Journal Title: Mod Rheumatol Volume: 21 Pages: 608-20
  • Peer Reviewed
• [Journal Article] G alpha(q) signal in osteoblasts is inhibitory to the osteoanabolic action of parathyroid hormone (2011)
  • Author(s): Ogata N, Kawaguchi H, et al.
  • Journal Title: J Biol Chem Volume: 286 Pages: 13733-40
  • Peer Reviewed
• [Journal Article] Ossification of the posterior atlantoaxial membrane associated with an os odontoideum : a case report (2011)
  • Author(s): Ohya J, Chikuda H, et al.
  • Journal Title: J Orthop Surg Volume: 19 Pages: 392-4
  • DOI: PMID:22184181
  • Peer Reviewed
• [Journal Article] Use of Doppler ultrasonography to detect an elusive communication of a spinal extradural arachnoid cyst (2011)
  • Author(s): Kanetaka M, Chikuda H, et al.
  • Journal Title: J Clin Neurosci Volume: 18 Pages: 863-4
  • DOI: PMID:21489797
  • Peer Reviewed
• [Journal Article] Acute cervical spinal cord injury complicated by preexisting ossification of the posterior longitudinal ligament : a multicenter study (2011)
  • Author(s): Chikuda H, et al.
  • Journal Title: Spine Volume: 36 Pages: 1453-8
  • DOI: PMID:21240049
  • Peer Reviewed
• [Presentation] NF-/κBファミリーRelAは、ADAMTS5の転写誘導因子として変形性関節症の進行に作用する (2012)
  • Author(s): 小林寛、川口浩, 他
  • Organizer: 日本軟骨代謝学会
  • Place of Presentation: 名古屋
  • Year and Date: 2012-09-03
• [Presentation] C/EBPb and RUNX2 Cooperatively Control Cartilage Degradation with MMP-13 as the Target and HIF-2a as the Inducer in Chondrocytes (2011)
  • Author(s): Hirata M, Kawaguchi H, et al.
  • Organizer: The American Society for Bone and Mineral Research (ASBMR)
  • Place of Presentation: San Diego, USA
  • Year and Date: 2011-09-17
• [Presentation] Notch/Rbpj/Hes1 Signal in Chondrocytes Modulates the Terminal Stage of Endochondral Ossification during Skeletal Growth and Osteoarthritis Development (2011)
  • Author(s): Hosaka Y, Kawaguchi H, et al.
  • Organizer: The American Society for Bone and Mineral Research (ASBMR)
  • Place of Presentation: San Diego, USA
  • Year and Date: 2011-09-17
• [Presentation] Functional Redundancy of GSK-3α and GSK-3β to Control Chondrocyte Differentiation through Phosphorylation of RelA/NF-κB p65 (2011)
  • Author(s): Itoh S, Kawaguchi H, et al.
  • Organizer: Osteoarthritis Research Society International (OARSI)
  • Place of Presentation: San Diego, USA
  • Year and Date: 2011-09-16
• [Presentation] Transcriptional induction of ADAMTS5 by NF-κB family member RelA/p65 in chondrocytes during osteoarthritis development (2011)
  • Author(s): Kobayashi H, Kawaguchi H, et al.
  • Organizer: International Osteoporosis Federation/Australian and New Zealand Bone & Mineral Society (IOF/ANZBMS)
  • Place of Presentation: Gold coast (AUS)
  • Year and Date: 2011-09-07
• [Presentation] NF-κBファミリーRelAは、ADAMTS5の転写誘導因子として変形性関節症の進行に作用する (2011)
  • Author(s): 小林寛、川口浩, 他
  • Organizer: 日本骨代謝学会
  • Place of Presentation: 大阪
  • Year and Date: 2011-07-30
• [Presentation] 軟骨細胞のNotch/Rbpj/Hes1シグナルは骨格形成および変形性関節症における軟骨内骨化の後期過程を制御する (2011)
  • Author(s): 保坂陽子、川口浩, 他
  • Organizer: 日本骨代謝学会
  • Place of Presentation: 大阪
  • Year and Date: 2011-07-30
• [Presentation] GSK-3αとGSK-3βは協調的にRelA/NF-κB p65のリン酸化を介して軟骨細胞初期分化を制御する (2011)
  • Author(s): 伊藤祥三、川口浩, 他
  • Organizer: 日本骨代謝学会
  • Place of Presentation: 大阪
  • Year and Date: 2011-07-29
• [Remarks]
  • URL: http://www.h.u-tokyo.ac.jp/ortho/