妊娠期に特有な後天性・血栓性微小血管障害症の分子病態とその制御機構解析

2014 Fiscal Year Annual Research Report

• Project/Area Number: 23591425
• Research Institution: Nara Medical University
• Principal Investigator: 藤村 吉博 奈良県立医科大学, 医学部, 名誉教授 (80118033)
• Co-Investigator(Kenkyū-buntansha): 松本 雅則 奈良県立医科大学, 医学部, 教授 (60316081) ; 佐道 俊幸 奈良県立医科大学, 医学部, 講師 (50275335) [Withdrawn] ; 早川 正樹 奈良県立医科大学, 医学部, 助教 (30516729)
• Project Period (FY): 2011-04-28 – 2015-03-31
• Keywords: 妊娠関連TMA / HELLP症候群 / ADAMTS13 / VWF / 胎盤由来E-NTPase

Outline of Annual Research Achievements

2013年末までに正常妊婦129例、妊娠高血圧症候群（PIH）24例、HELLP症候群10例の連続的検体の集積を行った。集積した妊婦血漿を用いて、ADAMTS13を等電点電気泳動（IEF）で解析した結果、HELLP症候群では“VWF非結合型ADAMTS13”の著明な欠損と“VWF結合型ADAMTS13”の増加を認め、これは症状の改善とともに正常化した。またPIHにおいても一部の重症例で軽度の“非結合型ADAMTS13”の欠損を認めた。我々は以前に“VWF非結合型ADAMTS13”は高ずり応力惹起凝集を初期より阻害し、“VWF結合型”は凝集を後期に阻害することを同定したことから（Hori et al, 2013）、HELLP症候群における微小血管障害の進展にはADAMTS13による早期の血小板凝集阻害能の減弱が関与する可能性が示唆された。
一方、申請者らは胎盤由来E-NTPaseが妊娠期の血小板血栓調節機構に関与しうる可能性を想定しE-NTPaseに対するモノクローナル抗体（mAb）を9種類作成した。各々のmAbがE-NTPase活性に与える影響を解析したところ、興味深いことに1種類のみ活性を助長するような挙動を示す抗体を同定した。作成したE-NTPaseのmAbを用いて血漿中E-NTPase抗原や活性を定量するELISA系を樹立したが、正常妊婦とHELLP症例の間で有意な差を見出すことが困難であったことから、今後はフローサイトメトリー等を用いた測定系を検討する。
従来、HELLP症候群におけるADAMTS13解析は同酵素の抗原量や活性にのみ焦点が当てられ、有意な関連性は見出されていなかった。本研究はIEFという申請者独自の手法を用いたことで、HELLP症候群におけるADAMTS13の血小板凝集阻害機能低下を見出した初の報告であり、本疾患の病態解明に貢献しうると考える。

Research Products

• [Journal Article] A unique case involving a female patient with Upshaw-Schulman syndrome: low titers of antibodies against ADAMTS13 prior to pregnancy disappeared after successful deliver. (2015)
  • Author(s): Ogawa Y, Matsumoto M, Sadakata H, Isonishi A, Kato S, Nojima Y, Fujimura Y.
  • Journal Title: Transfus Med Hemother. Volume: 42 Pages: 59-63
  • DOI: 10.1159/000370225.
  • Peer Reviewed
• [Journal Article] Eculizumab in the treatment of atypical hemolytic uremic syndrome in an infant leads to cessation of peritoneal dialysis and improvement of severe hypertension. (2015)
  • Author(s): Ohta T, Urayama K, Tada Y, Furue T, Imai S, Matsubara K, Ono H, Sakano T, Jinno K, Yoshida Y, Miyata T, Fujimura Y.
  • Journal Title: Pediatr Nephrol. Volume: 30 Pages: 603-608
  • DOI: 10.1007/s00467-014-2975-4.
  • Peer Reviewed
• [Journal Article] Familial C3 glomerulonephritis associated with mutations in the gene for complement factor B. (2015)
  • Author(s): Imamura H, Konomoto T, Tanaka E, Hisano S, Yoshida Y, Fujimura Y, Miyata T, Nunoi H.
  • Journal Title: Nephrol Dial Transplant. Volume: 30 Pages: 862-864
  • DOI: 10.1093/ndt/gfv054.
  • Peer Reviewed
• [Journal Article] Autoimmune-type atypical hemolytic uremic syndrome treated with eculizumab as first-line therapy. (2015)
  • Author(s): Hisano M, Ashida A, Nakano E, Suehiro M, Yoshida Y, Matsumoto M, Miyata T, Fujimura Y, Hattori M.
  • Journal Title: Pediatrics Int. Volume: 57 Pages: 313-317
  • DOI: 10.1111/ped.12469.
  • Peer Reviewed
• [Journal Article] STEC:O111-HUS complicated by acute encephalopathy in a young girl was successfully treated with a set of hemodiafiltration, steroid pulse, and soluble thrombomodulin, under plasma exchange. (2015)
  • Author(s): Yada N, Fujioka M, Bennett CL, Inoki K, Miki T, Watanabe A, Yoshida T, Hayakawa M, Matsumoto M, Fujimura Y.
  • Journal Title: Clin Case Rep. Volume: 3 Pages: 208-212
  • DOI: 10.1002/ccr3.196.
  • Peer Reviewed
• [Journal Article] A Novel Quantitative Hemolytic Assay Coupled with Restriction Fragment Length Polymorphisms Analysis Enabled Early Diagnosis of Atypical Hemolytic Uremic Syndrome and Identified Unique Predisposing Mutations in Japan. (2015)
  • Author(s): Yoshida Y, Miyata T, Matsumoto M, Shirotani-Ikejima H, Uchida Y, Ohyama Y, Kokubo T, Fujimura Y.
  • Journal Title: PLos One. Volume: 10 Pages: e0124655
  • DOI: 10.1371/journal.pone.0124655.
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] A rapid, fully automatic and highly sensitive ADAMTS13 gold particle immunoassay using a routine biochemistry analyser. (2015)
  • Author(s): Kato S, Tanaka M, Isonishi A, Matsumoto M, Samori T, Fujimura Y.
  • Journal Title: Brit J Haematol Volume: in press Pages: in press
  • DOI: 10.1111/bjh.13425.
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Poor-responder to plasma exchange therapy in acquired TTP is associated with ADAMTS13 inhibitor boosting: Visualization of an ADAMTS13-inhibitor complex, and its proteolytic clearance from plasma. (2015)
  • Author(s): Isonishi A, Bennett C.L., Plaimauer B, Scheiflinger F, Matsumoto M, Fujimura Y.
  • Journal Title: Transfusion Volume: in press Pages: in press
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Candidate gene analysis using genomic quantitative PCR: identification of ADAMTS13 large deletions in two patients with Upshaw-Schulman syndrome. (2014)
  • Author(s): Eura Y, Kokame K, Takafuta T, Tanaka R, Kobayashi H, Ishida F, Hisanaga S, Matsumoto M, Fujimura Y, Miyata T.
  • Journal Title: olecular Genetics & Genomic Medicine. Volume: 2 Pages: 240-244
  • DOI: 10.1002/mgg3.64.
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Analysis of patients with atypical hemolytic uremic syndrome treated at the Mie University Hospital: Concentration of C3 p.I1157T mutation. (2014)
  • Author(s): Matsumoto T, Fan X, Ishikawa E, Ito M, Amano K, Toyoda H, Komada Y, Ohishi K, Katayama N, Yoshida Y, Matsumoto M, Fujimura Y, Ikejiri M, Wada H, Miyata T.
  • Journal Title: Int J Hematol. Volume: 100 Pages: 437-442
  • DOI: 10.1007/s12185-014-1655-2.
  • Peer Reviewed
• [Journal Article] Postoperative atypical hemolytic uremic syndrome associated with complement c3 mutation. (2014)
  • Author(s): Matsukuma E, Imamura A, Iwata , Takeuchi T, Yoshida Y, Fujimura Y, Fan X, Miyata T, Kuwahara T.
  • Journal Title: Case Rep Nephrol. Volume: 2014 Pages: 784943
  • DOI: 10.1155/2014/784943.
  • Peer Reviewed
• [Presentation] TMA：最近の考え方と治療法の選択 (2015)
  • Author(s): 藤村吉博
  • Organizer: Nagoya Hematology Forum
  • Place of Presentation: ヒルトン名古屋（愛知県・名古屋市）
  • Year and Date: 2015-02-06
  • Invited
• [Presentation] Introduction of a quick assay for ADAMTS13 activity improved a survival of acquired TTP patients who received platelet transfusions. (2014)
  • Author(s): Yoshii Ｙ, Matsumoto M, Kurumatani N, Isonishi A, Uemura M, Hori Y, Hayakawa M, Yagi H, Bennett C.L., Fujimura Y
  • Organizer: The 56th Annual meeting of American Society of Hematology
  • Place of Presentation: San Francisco（USA）
  • Year and Date: 2014-12-08
• [Presentation] Decreased platelet thrombus size, due to a heightened proteolysis of VWF by ADAMTS13 is quickly restored after valve replacement in aortic stenosis patients. (2014)
  • Author(s): Yagi H, Hayakawa M, Yamaguchi N, Yamashita K, Taniguchi S, Matsumoto M, Tsubaki K, Fujimura Y
  • Organizer: The 56th Annual meeting of American Society of Hematology
  • Place of Presentation: San Francisco（USA）
  • Year and Date: 2014-12-07
• [Presentation] Registry of congenital thrombotic microangiopathy in Japan. (2014)
  • Author(s): Fujimura Y
  • Organizer: The 3rd ASEAN Federation of Hematology Congress.Plenary lecture
  • Place of Presentation: Bangkok（Thailand）
  • Year and Date: 2014-10-23
  • Invited
• [Presentation] ADAMTS13と血小板輸血-過去、現在、未来 (2014)
  • Author(s): 藤村吉博
  • Organizer: 第20回 ADAMTS13研究会
  • Place of Presentation: 奈良ホテル（奈良県・奈良市）
  • Year and Date: 2014-09-18
  • Invited
• [Presentation] TTP, HUS, aHUSの鑑別診断と治療. (2014)
  • Author(s): 藤村吉博
  • Organizer: 第59回臨床病理近畿支部例会
  • Place of Presentation: 天理医療大学キャンパス（奈良県・天理市）
  • Year and Date: 2014-06-21
  • Invited
• [Book] ADAMTS13 book (2015)
  • Author(s): Fujimura Y, Kokame K, Yagi H, Isonishi A, Matsumoto M, Miyata T
  • Total Pages: in press
  • Publisher: Springer (edited by George Rodgers)