人工関節置換術術後の静脈血栓塞栓症発症関連遺伝子と抗凝固薬反応関連遺伝子の同定

• Project/Area Number: 20791057
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Single-year Grants
• Research Field: Orthopaedic surgery
• Research Institution: Tokyo Women's Medical University
• Principal Investigator: 塚原 聡 東京女子医科大学, 医学部, 助教 (90433995)
• Project Period (FY): 2008 – 2010
• Project Status: Completed (Fiscal Year 2010)
• Budget Amount: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000); Fiscal Year 2010: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2009: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2008: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: 人工関節 / 静脈血栓 / ゲノム / 薬剤反応性 / 疾患感受性

Research Abstract

術後VTE発症予測遺伝子およびフォンダパリヌクス反応性関連遺伝子の各候補遺伝子について遺伝子多型解析を行うため、手術患者ほぼ全例にDNA採取に対する協力を求め、同意を得られた患者からはインフォームドコンセントの下にDNA採取を行い、厳格な連結可能匿名化の上で手術データベースと連動させた。
アミノ酸非同義置換を起こすSNPは、タンパクの親和性を含めた性質変換の可能性がありGeneCards(http://www.genecards.org/index.shtml)、ExPasy(Expert Protein Analysis System)Proteomics Server(http://us.expasy.org/)データベースを利用して遺伝子のモチーフやトランスクリプトバリアント、タンパク配列と構造を検索し、目的遺伝子の機能解析を行った。エクソン領域のアミノ酸非同義置換を起こすSNPは免疫共沈降法(IP;Immunopr ecipitation assay)により標的分子の親和性を検討した。イントロン領域およびプロモーター領域、5'あるいは3'UTRに存在するSNPは、TESS(Transcription Element Search System)データベースにより転写因子の結合を確認し、TaqMan法を利用したReal-time quantitative PCRによる発現解析、目的のSNPを含有したルシフェラーゼアッセイによるプロモーター活性解析、SNPを含むゲノムDNAとタンパクとの結合を確認するElectrophoretic Mobility Shift Assay(EMSA)の実験を行った。同時に第Xa凝固因子活性の検討のために、標的分子およびATIII、フォンダパリヌクス、Xa、発色基質を同時にインキュベートし、吸光度測定して基質分解を定量化した。また転写活性に関連する領域以外のSNPの場合には遺伝子多型に依存する当施設で採取しているB cell lineのmRNA変異の解析を行った。

Research Products

• [Journal Article] Association study of TRAFI-C5 polymorphisms with susceptibility to rheumatoid arthritis and systemic lupus erythematosus in Japanese. (2009)
  • Author(s): Tsukahara, et al.
  • Journal Title: Ann Rheum Dis Epub ahead of print
  • Peer Reviewed
• [Journal Article] Decrease in orthopaedic surgeries including total joint replacements in rheumatoid arthritis patients between 2001 and 2007 : data from Japanese outpatients in a single institute-based large observational cohort (IORRA). (2009)
  • Author(s): Tsukahara, et al.
  • Journal Title: Ann Rheum Dis 69 Pages: 312-313
  • Peer Reviewed
• [Journal Article] RA患者に対するセメントレスTKAの中期成績 (2009)
  • Author(s): 塚原, ら
  • Journal Title: 日本関節病学会誌 28 Pages: 59-64
  • NAID: 130004567879
  • Peer Reviewed
• [Journal Article] Q&A関節リウマチ患者の人工関節置換術を整形外科に依頼するポイント (2009)
  • Author(s): 塚原, ら
  • Journal Title: Frontiers in Rheumatology & Clinical Immunology 3 Pages: 166-166
• [Journal Article] Declining use of synovectomy surgery for patients with rheumatoid arthritis in Japan (2009)
  • Author(s): Tsukahara, et.al.
  • Journal Title: Ann Rheum Dis 68 Pages: 291-292
  • Peer Reviewed
• [Journal Article] Return of infliximab efficacy after total knee arthroplasty in a patient with rheumatoid arthritis (2008)
  • Author(s): Tsukahara, et.al.
  • Journal Title: Clin Rheumatol 27 Pages: 549-550
  • Peer Reviewed
• [Journal Article] Iliopsoas bursitis-associated femoral neuropathy exacerbated after internal fixation of an intertrochanteric hip fracture in rheumatoid arthritis : a case report. (2008)
  • Author(s): Tsukahara, et.al.
  • Journal Title: Mod Rheumatol 18 Pages: 394-398
  • Peer Reviewed
• [Journal Article] Risk factors for wrist surgery in rheumatoid arthritis. (2008)
  • Author(s): Tsukahara, et.al.
  • Journal Title: Clin Rheumatol 27 Pages: 1387-1391
  • Peer Reviewed
• [Journal Article] Association of STAT4 with susceptibility to rheumatoid arthritis and systemic lupus ervthematosus in the Japanese population. (2008)
  • Author(s): Tsukahara, et.al.
  • Journal Title: Arthritis Rheum. 58 Pages: 1940-1946
  • Peer Reviewed
• [Journal Article] Infusion reaction to infliximab in a patient with rheumatoid arthritis after discontinuation over 1 year and readministration (2008)
  • Author(s): Tsukahara, et.al.
  • Journal Title: J Rheumatol 35 Pages: 1896-1897
  • Peer Reviewed
• [Journal Article] A polymorphism in the gene encoding the Fcgamma IIIA receptor is a possible genetic marker to predict the primary response to infliximab in Japanese patients with rheumatoid arthritis (2008)
  • Author(s): Tsukahara, et.al.
  • Journal Title: Ann Rheum Dis 67 Pages: 1791-1792
  • Peer Reviewed
• [Presentation] Effect of CYP2C9 and VKORC1 Genotypes On Early-Phase Warfarin Dosing of Rheumatoid Arthritis Patients with Elective Joint Surgery in Japanese (2009)
  • Author(s): Tsukahara, et al.
  • Organizer: ACR/ARHP Annual Scientific Meeting
  • Place of Presentation: Philadelphia, USA
  • Year and Date: 2009-10-20
• [Presentation] Perioperative Complications in Patients with RA Receiving Tumor Necrosis Factor Inhibitor (2009)
  • Author(s): Tsukahara, et al.
  • Organizer: ACR/ARHP Annual Scientific Meeting
  • Place of Presentation: Philadelphia, USA
  • Year and Date: 2009-10-20
• [Presentation] Interleukin 2 Receptor, Alpha Is Associated with Radiographic Joint Damage in a Japanese RA Population (2009)
  • Author(s): Tsukahara, et al.
  • Organizer: ACR/ARHP Annual Scientific Meeting
  • Place of Presentation: Philadelphia, USA
  • Year and Date: 2009-10-19
• [Presentation] 日本リウマチ学会 (2009)
  • Author(s): 塚原, ら
  • Organizer: RA患者に対するTKA術後のフォンダパリヌクスとエノキサパリンの効果と安全性
  • Place of Presentation: 東京
• [Presentation] 関節リウマチに対する下肢人工関節置換術時のフォンダパリヌクスの安全性の検討 (2009)
  • Author(s): 塚原, ら
  • Organizer: 日本整形外科学会
  • Place of Presentation: 福岡
• [Presentation] Replication study for the Wellcome Trust Case Control Consortium data in large-scaled Japanese rheumatoid arthritis case-control populations (2008)
  • Author(s): Tsukahara, et.al.
  • Organizer: The American Society of Human Genetics 58th Annual Meeting
  • Place of Presentation: Philadelphia
  • Year and Date: 2008-11-14
• [Presentation] Association of nine novel polymorphisms to rheumatoid arthritis susceptibility, identified by the Wellcome Trust Case Control Consortium, with Japanese rheumatoid arthritis patients (2008)
  • Author(s): Tsukahara, et.al.
  • Organizer: American College of Rheumatology
  • Place of Presentation: San Francisco
  • Year and Date: 2008-10-26
• [Presentation] 6q23領域に存在する遺伝子多型と日本人関節リウマチとの関連 (2008)
  • Author(s): 塚原聡
  • Organizer: 日本整形外科学会基礎学術集会
  • Place of Presentation: 京都
  • Year and Date: 2008-10-24
• [Presentation] ASSOCIATION OF A POLYMORPHISM ON 6Q23 WITH RHEUMATOID ARTHRITIS IN A JAPANESE POPULATION (2008)
  • Author(s): Tsukahara, et.al.
  • Organizer: The Annual European Congress of Rheumatology EULAR 2008
  • Place of Presentation: Paris
  • Year and Date: 2008-06-14
• [Presentation] WeHcome Trust Case Control Consortiumによって同定された9個の関節リウマチ新規感受性遺伝子多型と日本人関節リウマチとの関連 (2008)
  • Author(s): 塚原聡
  • Organizer: 日本リウマチ学会
  • Place of Presentation: 札幌
  • Year and Date: 2008-04-21
• [Remarks]
  • URL: http://web.me.com/twmu/JointSurgery/Home.html