Pathophysiology and targeted treatment of autoimmune hematological diseases based on the next generation high-throughput TCR/BCR repertoire analysis

• Project/Area Number: 15K08639
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Laboratory medicine
• Research Institution: Akita University
• Principal Investigator: Hirokawa Makoto 秋田大学, 医学(系)研究科(研究院), 教授 (50241667)
• Co-Investigator(Kenkyū-buntansha): 鈴木 隆二 独立行政法人国立病院機構(相模原病院臨床研究センター), 診断・治療研究室, 室長 (70373470) ; 藤島 直仁 秋田大学, 医学部, 講師 (70422152)
• Co-Investigator(Renkei-kenkyūsha): KITAURA Kazutaka 国立相模原病院, 診断治療研究室, 研究員 (00518136) ; MATSUTANI Takaji 国立相模原病院, 診断治療研究室, 研究員 (70372290)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2017: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2015: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: 特発性血小板減少性紫斑病 / Bリンパ球 / B細胞抗原受容体 / レパトア / NGS / 次世代シーケンサー / ハイスループット・レパトア解析 / T細胞抗原受容体 / 赤芽球癆 / リンパ系腫瘍

Outline of Final Research Achievements

To identify the pathological B-cell clones in primary ITP, we have investigated the repertoires of IgG-BCRs of peripheral blood B cells by using the high-throughput DNA sequencing. Unbiased amplifications of IgG-BCRs were performed by adaptor-ligation PCR. The amplicons were sequenced by next-generation sequencing. Comparisons of the IGHV repertoires covering fifty-nine IGHV subfamilies between the patients and controls revealed an increased usage of IGHV4-28 in ITP patients. The 186 distinct IGHV4-28-carrying sequences were identified in ITP patients and 135 out of these clones used the IGHJ4 gene segment. The present study has demonstrated the preferential usage of the rearranged IGHV4-28/IGHJ4 gene by circulating IgG B cells in ITP. Although the antigen specificity of BCRs encoded by the IGHV4-28/IGHJ4 gene in the present patient cohort remains to be elucidated, prolonged antigenic pressure appears to shape this characteristic BCR repertoire.

Research Products

• [Journal Article] 赤芽球癆：疾患概念・病因・病態 (2017)
  • Author(s): 廣川 誠
  • Journal Title: 日本臨牀 Volume: 75 Pages: 431-436
• [Journal Article] 後天性赤芽球癆の特徴と治療の実際 (2017)
  • Author(s): 廣川 誠
  • Journal Title: 新薬と臨床 Volume: 66 Pages: 62-67
• [Journal Article] 成人大球性貧血の診断 (2017)
  • Author(s): 廣川 誠
  • Journal Title: Medical Practice Volume: 34 Pages: 1213-1213
• [Journal Article] Diagnosis and treatment of macrocytic anemias in adults (2017)
  • Author(s): Nagao, T., Hirokawa, M.
  • Journal Title: J Gen Fam Med Volume: 18 Issue: 5 Pages: 200-204
  • DOI: 10.1002/jgf2.31
  • Peer Reviewed
• [Journal Article] Reduced-intensity conditioning allogeneic hematopoietic cell transplantation for younger patients with acute myeloid leukemia: a registry-based study (2017)
  • Author(s): Yanada, M., Kurosawa, S., Kobayashi, T., Ozawa, Y., Kanamori, H., Kobayashi, N., Sawa, M., Nakamae, H., Uchida, N., Hashimoto, H., Fukuda, T., Hirokawa, M., Atsuta, Y., Yano, S.
  • Journal Title: Bone Marrow Transplant Volume: 52 Issue: 6 Pages: 818-824
  • DOI: 10.1038/bmt.2016.358
• [Journal Article] Progress in the clinical management of pure red cell aplasia and future prospects (2016)
  • Author(s): 廣川 誠
  • Journal Title: Rinsho Ketsueki Volume: 57 Issue: 2 Pages: 110-116
  • DOI: 10.11406/rinketsu.57.110
  • NAID: 130005132709
  • ISSN: 0485-1439, 1882-0824
  • Peer Reviewed
• [Journal Article] 臨床検査技師のスキルアップを考える. 臨床病理 64(1)，89-95 (2016)
  • Author(s): 小林則子，鎌田由美子，富谷陽子，菊地優子，面川 歩，嵯峨知生，植木重治，廣川 誠
  • Journal Title: 臨床病理 Volume: 64 Pages: 89-95
  • Peer Reviewed
• [Journal Article] Impact of age on outcomes of allogeneic hematopoietic stem cell transplantation with reduced intensity conditioning in elderly patients with acute myeloid leukemia (2016)
  • Author(s): Aoki J，Kanamori H，Tanaka M，Yamasaki S，Fukuda T，Ogawa H，Iwato K，Ohashi K，Okumura H，Onizuka M，Maesako Y，Teshima T，Kobayashi N，Morishima Y，Hirokawa M，Atsuta Y，Yano S，Takami A
  • Journal Title: Am J Hematol Volume: 91 Issue: 3 Pages: 302-307
  • DOI: 10.1002/ajh.24270
  • Peer Reviewed
• [Journal Article] Cytosine-phosphorothionate- guanine oligodeoxynucleotides exacerbates hemophagocytosis by inducing tumor necrosis factor-α production in mice after bone marrow transplanation. (2016)
  • Author(s): Liu J, Guo YM, Onai N, Ohyagi H, Hirokawa M, Takahashi N, Tagawa H, Ubukawa K, Kobayahi I, Tezuka H, Minamiya Y, Ohteki T, and Sawada K.
  • Journal Title: Biol Blood Marrow Transplant 2015 Volume: 印刷中 Issue: 4 Pages: 627-636
  • DOI: 10.1016/j.bbmt.2015.12.018
  • Peer Reviewed / Open Access
• [Journal Article] L-アスパラギナーゼ治療中のアンモニア測定に関する検査室の経験：採血管内での上昇について (2016)
  • Author(s): 高橋 縁，藤田美好，植木重治，廣川 誠
  • Journal Title: 臨床化学 Volume: 45 Pages: 269-273
  • Peer Reviewed
• [Journal Article] Eosinophil ETosis and DNA Traps: a New Look at Eosinophilic Inflammation. (2016)
  • Author(s): Ueki S, Tokunaga T, Fujieda S, Honda K, et al
  • Journal Title: Curr Allergy Asthma Rep. Volume: 16(8) Issue: 8 Pages: 1-17
  • DOI: 10.1007/s11882-016-0634-5
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] 赤芽球癆の発症機序と診断の進めかた (2016)
  • Author(s): 廣川 誠
  • Journal Title: Medical Practice Volume: 33 Pages: 1413-1417
  • Peer Reviewed
• [Journal Article] 脱顆粒を伴う好酸球の非アポトーシス細胞死 (2016)
  • Author(s): 植木重治，本田耕平，廣川 誠
  • Journal Title: アレルギーの臨床 Volume: 36 Pages: 71-73
  • Peer Reviewed
• [Journal Article] 好酸球性炎症の新しい機序 (2016)
  • Author(s): 植木重治,中村由夏,廣川 誠
  • Journal Title: アレルギーの臨床 Volume: 36 Pages: 1349-1352
  • Peer Reviewed
• [Journal Article] Eosinophil extracellular trap cell death-derived DNA traps: Their presence in secretions and functional attributes. (2016)
  • Author(s): Ueki S, Konno Y, Takeda M, Moritoki Y, Hirokawa M, Matsuwaki Y, Honda K, Ohta N, Yamamoto S, Takagi Y, Wada A, Weller PF.
  • Journal Title: J Allergy Clin Immunol. Volume: 137 Issue: 1 Pages: 258-267
  • DOI: 10.1016/j.jaci.2015.04.041
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Possible involvement of reusable towels in the high rate of Bacillus species-positive blood cultures in Japanese hospitals (2016)
  • Author(s): Saito N, Kondo J, Haruki S, Itoga M, Yamamoto A, Kim ura M, Inoue F, Kobayashi M, Tsutaya S, Kojima K, Ueki S, Hirokawa M, Kayaba H
  • Journal Title: J Infect Chemother Volume: 22 Issue: 2 Pages: 96-101
  • DOI: 10.1016/j.jiac.2015.11.006
  • Peer Reviewed
• [Journal Article] The effect of hepatocyte growth factor on secretory functions in human eosinophils (2016)
  • Author(s): Yamauchi Y, Ueki S, Konno Y, Ito W, Takeda M, Nakamura Y, Nishikawa J, Moritoki Y, Omokawa A, Saga T, Hirokawa M
  • Journal Title: Cytokine Volume: 88 Pages: 45-50
  • DOI: 10.1016/j.cyto.2016.08.013
  • Peer Reviewed
• [Journal Article] Long-term outcome of patients with acquired chronic pure red cell aplasia (PRCA) following immunosuppressive therapy: a final report of the nationwide cohort study in 2004/2006 by the Japan PRCA collaborative study group. (2015)
  • Author(s): Hirokawa M, Sawada K, Fujishima N, Teramura M, Bessho M, Dan K, Tsurumi H, Nakao S, Urabe A, Fujisawa S, Yonemura Y, Kawano F, Oshimi K, Sugimoto K, Matsuda A, Karasawa M, Arai A, Komatsu N, Harigae H, Omine M, Ozawa K, Kurokawa M; PRCA Collaborative Study Group.
  • Journal Title: Br J Haematol. Volume: in press Issue: 6 Pages: 879-886
  • DOI: 10.1111/bjh.13376
  • Peer Reviewed / Open Access
• [Journal Article] Functional ana- lysis of free fatty acid receptor GPR120 in human eosinophils: implications in metabolic homeostasis. (2015)
  • Author(s): Konno, Y., Ueki, S., Takeda, M., Kobayashi, Y., Tamaki, M., Moritoki, Y., Oyamada, H., Itoga, M., Kayaba, H., Omokawa, A., Hirokawa, M.
  • Journal Title: PLoS One Volume: 10 Issue: 3 Pages: e0120386-e0120386
  • DOI: 10.1371/journal.pone.0120386
  • Peer Reviewed / Open Access
• [Journal Article] G-Protein-Coupled Estrogen Receptor Agonist Suppresses Airway Inflammation in a Mouse Model of Asthma through IL-10. (2015)
  • Author(s): Itoga, M., Konno, Y., Moritoki, Y., Saito, Y., Ito, W., Tamaki, M., Kobayashi, Y., Kayaba, H., Kikuchi, Y., Chihara, J., Takeda, M., Ueki, S., Hirokawa, M.
  • Journal Title: PLoS One Volume: 10 Issue: 3 Pages: e0123210-e0123210
  • DOI: 10.1371/journal.pone.0123210
  • Peer Reviewed / Open Access
• [Journal Article] 再生不良性貧血と赤芽球癆の病態と治療 (2015)
  • Author(s): 廣川 誠，藤島直仁，面川 歩，植木重治
  • Journal Title: 日本内科学会雑誌 Volume: 104 Pages: 1405-1413
• [Journal Article] Diagnosis and management of pure red cell aplasia (2015)
  • Author(s): 廣川 誠
  • Journal Title: Rinsho Ketsueki Volume: 56 Issue: 10 Pages: 1922-1931
  • DOI: 10.11406/rinketsu.56.1922
  • NAID: 130005103308
  • ISSN: 0485-1439, 1882-0824
• [Presentation] 後天性慢性赤芽球癆の長期予後を明らかにするための多施設共同前向き観察研究(PRCA2016)中間報告 (2017)
  • Author(s): 廣川 誠，藤島直仁，澤田賢一，張替秀郎，松田 晃，小松則夫，通山 薫，米村雄士，中尾眞二，齋藤明子，松村 到，荒井俊也，黒川峰夫
  • Organizer: 第79回日本血液学会学術集会
• [Presentation] Identification of Adverse Risk Factors for Survival in Acquired Pure Red Cell Aplasia Receiving Immunosuppressive Therapy By the Japan PRCA2004/2006 Study and Introduction to the New Cohort Study PRCA2016 (2017)
  • Author(s): Hirokawa, M., Fujishima, N., Teramura, M., et al.
  • Organizer: 59th Annual Meeting of American Society of Hematology
  • Int'l Joint Research
• [Presentation] High-throughput sequencing of IgG B-cell receptors reveals frequent usage of the rearranged IGHV4/IGHJ4 gene in primary immune thrombocytopenia in adults (2017)
  • Author(s): Fujishima, N., Togashi, M., Hasegawa, R., Watanabe, K., Saga, A., Omokawa, A., Saga, T., Ueki, S., Takahashi, N., Kitaura, K., Suzuki, R., Hirokawa, M.
  • Organizer: 59th Annual Meeting of American Society of Hematology
  • Int'l Joint Research
• [Presentation] Comparison of the number of genome-wide single nucleotide polymorphisms (SNPs) among epidemiologically-related and -unrelated methicillin-resistant Staphylococcus aureus (MRSA) isolates of indistinguishable genotype by PCR-based open-reading frame typing (POT) kit (2016)
  • Author(s): 1)Saga T, Tatsuko R, Aoki K, Omokawa A, Moritoki Y, Shimizu-Saga A, Kobayashi N, Ueki S, Ishii Y, Tateda K, and Hirokawa M
  • Organizer: IDWeek 2016
  • Place of Presentation: New Orleans, USA
  • Year and Date: 2016-10-26
  • Int'l Joint Research
• [Presentation] 好酸球性ムチンの粘性はなぜ高いのか－好酸球と好中球DNA trapsの差異 (2016)
  • Author(s): 植木重治，中村由夏，今野泰典，竹田正秀，守時由起，廣川 誠，本田耕平，太田伸男，松脇良典，Peter Weller
  • Organizer: 第65回日本アレルギー学会学術大会
  • Place of Presentation: 東京
  • Year and Date: 2016-06-17
• [Presentation] 5)Itoga M, Konno Y，Moritoki Y，Saito Y，Ito W, Tamaki M, Kobayashi Y, Kayaba H, Kikuchi Y, Chihara J, Takeda M, Ueki S, Hirokawa M (2016)
  • Author(s): Itoga M, Konno Y，Moritoki Y，Saito Y，Ito W, Tamaki M, Kobayashi Y, Kayaba H, Kikuchi Y, Chihara J, Takeda M, Ueki S, Hirokawa M
  • Organizer: 第65回日本アレルギー学会学術大会
  • Place of Presentation: 東京
  • Year and Date: 2016-06-17
• [Presentation] Recent updates in acquired pure red cell aplasia in the adults (2016)
  • Author(s): Hirokawa M
  • Organizer: Taiwan-Japan Hematology Forum HST-EHA Joint Symposium
  • Place of Presentation: Taipei, Taiwan
  • Year and Date: 2016-04-16
  • Int'l Joint Research / Invited
• [Presentation] 好酸球と好中球のETosisにより生じるDNA trapsの差異 (2016)
  • Author(s): 植木重治，今野泰典，竹田正秀，中村由夏，守時由起，面川 歩，嵯峨知生，廣川 誠，Peter Weller
  • Organizer: 第56回日本呼吸器学会
  • Place of Presentation: 京都
  • Year and Date: 2016-04-08
• [Presentation] ヒト好酸球に対する肝細胞増殖因子(HGF)の細胞外分泌能とETosisへの影響 (2016)
  • Author(s): 山内由美子，植木重治，中村由夏，今野泰典，面川歩，守時由起，伊藤亘，竹田正秀，嵯峨知生，廣川誠
  • Organizer: 第56回日本呼吸器学会
  • Place of Presentation: 京都
  • Year and Date: 2016-04-08
• [Presentation] Long-term outcome of patients with acquired chronic PRCA following immunosuppressive therapy (2015)
  • Author(s): Hirokawa, M., Sawada, K., Fuishima, N., Teramura, M., Bessho, M., Dan, K., Tsurumi, H., Nakao, S., Urabe, A., Fujisawa, S., Yonemura, Y., Kawano, F., Oshimi, K., Sugumoto, K., Matsuda, A., Karasawa, M., Arai, A., Komatsu, N., Harigae, H., Omine, M., Ozawa, K., Kurokawa, M.
  • Organizer: 第77回日本血液学会学術集会
  • Place of Presentation: 金沢
  • Year and Date: 2015-10-16
• [Presentation] 赤芽球癆の診断と治療（教育講演） (2015)
  • Author(s): 廣川 誠
  • Organizer: 第77回日本血液学会学術集会
  • Place of Presentation: 金沢
  • Year and Date: 2015-10-16
  • Invited
• [Presentation] TLR9 signaling exerts HPS in BMT mice and is effectively treated through TNF-α inhibition (2015)
  • Author(s): Liu, J., Guo, Y., Hirokawa, M., Takahasi, N., Ubukawa, K., Kobayashi, I., Minamiya, Y., Tezuka, H., Onai, N., Ohteki, T., Sawada, K.
  • Organizer: 第77回日本血液学会学術集会
  • Place of Presentation: 金沢
  • Year and Date: 2015-10-16
• [Presentation] Impact of donor source on allogeneic HSCT outcomes for patients with CMML. (2015)
  • Author(s): Itonaga, H., Aoki, K., Aoki, J., Ishiyama, K., Ishikawa, T., Uchida, N., Hoshino, T., Ohashi, K., Kurokawa, M., Ozawa, Y., Matsuoka, K., Yuiri, T., Hirokawa, M., Morishima, Y., Kato, K., Atsuta, Y., Miyazaki, Y.
  • Organizer: 第77回日本血液学会学術集会
  • Place of Presentation: 金沢
  • Year and Date: 2015-10-16
• [Presentation] Influence of CYP3A5 gene polymorphisms on pharmacokinetics of modified-release tacrolimus. (2015)
  • Author(s): Yamashita, T., Fujishima, N., Miura, M., Niioka, T., Abumiya, M., Nara, M., Fujishima, M., Kameoka, Y., Hirokawa, M., Takahashi, N.
  • Organizer: 第77回日本血液学会学術集会
  • Place of Presentation: 金沢
  • Year and Date: 2015-10-16
• [Book] 赤芽球癆．血液疾患最新の治療（小澤敬也編） (2017)
  • Author(s): 廣川 誠
  • Publisher: 南江堂
• [Book] 赤芽球癆．1336専門家による私の治療. 2017-18年度版（猿田亨男編） (2017)
  • Author(s): 廣川 誠
  • Publisher: 日本医事新報社
• [Book] 赤芽球癆. 貧血症 診断と治療のABC125（桐戸敬太編) (2017)
  • Author(s): 廣川 誠
  • Publisher: 最新医学社
• [Book] 慢性赤芽球癆の病態と治療、Annual Review2016血液（高久史麿、他編） (2016)
  • Author(s): 廣川 誠
  • Publisher: 中外医学社
• [Book] 赤芽球癆、血液科研修ノート（永井良三、神田善伸編） (2016)
  • Author(s): 3)廣川 誠
  • Publisher: 診断と治療社
• [Book] 血液専門医テキスト改訂第2版 (2015)
  • Author(s): 廣川 誠
  • Total Pages: 584
  • Publisher: 南江堂
• [Book] 今日の治療指針2015 (2015)
  • Author(s): 廣川 誠
  • Total Pages: 1973
  • Publisher: 医学書院
• [Book] 最新ガイドライン準拠血液疾患診断・治療指針 (2015)
  • Author(s): 廣川 誠
  • Total Pages: 560
  • Publisher: 中山書店
• [Book] ブラッシュアップ骨髄不全症 (2015)
  • Author(s): 廣川 誠
  • Total Pages: 193
  • Publisher: 中外医学社
• [Book] 免疫症候群 第2版Ⅰ（その他の免疫疾患を含めて） (2015)
  • Author(s): 廣川 誠，澤田賢一
  • Total Pages: 931
  • Publisher: 日本臨床社