Study on molecular pathogenesis of X-linked neutropenia and WIP deficiency by using NOG mice and human myeloid cell line.

• Project/Area Number: 23591528
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Pediatrics
• Research Institution: Tohoku University
• Principal Investigator: SASAHARA Yoji 東北大学, 医学(系)研究科(研究院), 准教授 (60372314)
• Project Period (FY): 2011-04-28 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2013: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2012: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2011: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: WASP / WIP / 原発性免疫不全症 / ヒト化マウス / NOGマウス / Wiskott-Aldrich症候群

Outline of Final Research Achievements

Rare gain-of-function mutations in the WASP gene are known to result in X-linked neutropenia (XLN). We first tried transdution of mutant WASP and WIP genes into CD34 positive human hematopoietic stem cells. However, transduction efficiency was very low. Therefore we next performed experiments by using myeloid cell line K562.
We reported that a part of WASP localized in the nucleus and a tendency of activating WASP mutants to localize in the nucleus more than WT. We found that WASP could form a complex with RNA polymerase II. To determine whether gene transcription was affected by WASP mutants in myeloid cells, we performed microarray analysis and found different expression profiles between WT and L270P WASP-transfected K562 cells. Among the genes affected, granulocyte colony-stimulating factor receptor(G-CSFR) and Runx1 were included. Therefore, our results suggested that open conformation of WASP would contribute to the outcome of myeloid cell differentiation and pathogenesis of XLN.

Research Products

• [Journal Article] Open conformation of WASP regulates its nuclear localization and gene transcription in myeloid cells. (2014)
  • Author(s): Looi CY, Sasahara Y, Watanabe Y, Satoh M, Hakozaki I, Du W, Uchiyama T, Kumaki S, Kure S, Tsuchiya S.
  • Journal Title: Int. Immunol. Volume: 26(6) Pages: 341-352
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Binding of WASP/N-WASP interacting protein WIP to actin regulates focal adhesion assembly and adhesion. (2014)
  • Author(s): Ramesh N, Massaad MJ, Kumar L, Suresh K, Sasahara Y, Anton I, Bhasin M, Libermann T, Geha RS.
  • Journal Title: Mol. Cell. Biol. Volume: 34(14) Pages: 2600-2610
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] GNE myopathy associated with congenital thrombocytopenia: a report of two siblings. (2014)
  • Author(s): Izumi R, Niihori T, Suzuki N, Sasahara Y, Nishiyama A, Nishiyama S, Endo K, Kato M, Warita H, Konno H, Tateyama M, Nagashima T, Funayama R, Nakayama K, Kure S, Matsubara Y, Aoki Y, Aoki M.
  • Journal Title: Neuromuscul. Disord. Volume: 24(12) Issue: 12 Pages: 1068-1072
  • DOI: 10.1016/j.nmd.2014.07.008
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] 総説 先天性血小板減少症の診断と分子病態 (2014)
  • Author(s): 笹原洋二
  • Journal Title: 小児科 Volume: 55(1) Pages: 105-114
• [Journal Article] A nationwide survey of Aicardi-Goutieres syndrome patients identifies a strong association between dominant TREX1 mutations and chilblain lesions: Japanese cohort study. (2014)
  • Author(s): Abe J, Nakamura K, Nishikomori R, Kato M, Mitsuiki N, Izawa K, Awaya T, Kawai T, Yasumi T, Toyoshima I, Hasegawa K, Ohshima Y, Hiragi T, Sasahara Y, Suzuki Y, Kikuchi M, Osaka H, Ohya T, Ninomiya S, Fujikawa S, Akasaka M, Iwata N, Kawakita A, Funatsuka M, Shintaku H, Ohara O, Ichinose H, Heike T.
  • Journal Title: Rheumatology Volume: 53(3) Pages: 448-458
  • Peer Reviewed
• [Journal Article] Selective expansion of regulatory T cells in mixed chimera after allogeneic bone marrow transplantation in a patient with IPEX syndrome. (2014)
  • Author(s): Horino S, Sasahara Y, Satoh M, Niizuma H, Kumaki S, Abukawa D, Satoh A, Imaizumi M, Kanegane H, Kamachi Y, Sasaki S, Terui K, Ito Y, Kobayashi I, Ariga T, Tsuchiya S, Kure S.
  • Journal Title: Pediatr. Transplant. Volume: 188(1)
  • Peer Reviewed
• [Journal Article] IDH2 and TP53 mutations are correlated with gliomagenesis in a patient with Maffucci syndrome. (2014)
  • Author(s): Moriya M, Kato Kaneko M, Liu X, Hosaka M, Fujishima F, Sakuma J, Ogasawara S, Watanabe M, Sasahara Y, Kure S, Kato Y.
  • Journal Title: Cancer Science Volume: 105(3) Pages: 359-362
  • Peer Reviewed
• [Journal Article] Mesenchymal chondrosarcoma diagnosed by FISH for HEY1-NCOA2 fusion gene. (2014)
  • Author(s): Moriya K, Katayama S, Onuma M, Hosaka M, Watanabe M, Hasegawa T, Sasahara Y, Kure S.
  • Journal Title: Pediatr. Int. Volume: in press.
  • Peer Reviewed
• [Journal Article] Unilateral phrenic nerve pulsy: A rare manifestation of vincristine neurotoxicity. (2014)
  • Author(s): Moriya K, Kakisaka Y, Onuma M, Sasahara Y, Kure S.
  • Journal Title: Indian J Pediatr. Volume: in press.
  • Peer Reviewed
• [Journal Article] Wiskott-Aldrich syndrome presenting with a clinical picture mimicking juvenile myelomonocytic leukemia. (2013)
  • Author(s): Oshimi A, Kamachi Y, Imai K, Watanabe N, Kinoshita A, Nakadate H, Kanazawa T, Oozono S, Kobayashi R, Yoshida M, Kobayashi C, Hama A, Muramatsu H, Sasahara Y, Jakob M, Morio T, Nonoyama S, Ehl S, Manabe A, Niemeyer C, Kojima S.
  • Journal Title: Pediatr. Blood & Cancer Volume: 60(5) Pages: 836-841
  • Peer Reviewed
• [Journal Article] Long-term outcome of childhood aplastic anemia patients who underwent allogeneic hematopoietic stem cell transplantation from HLA-matched sibling dono in Japan. (2013)
  • Author(s): Kikuchi A, Yabe H, Kato K, Koh K, Inagaki J, Sasahara Y, Suzuki R, Yoshida N, Kudo K, Kobayashi R, Tabuchi K, Kawa K, Kojima S.
  • Journal Title: Bone Marrow Transplant. Volume: 48(5) Pages: 657-660
  • Peer Reviewed
• [Journal Article] Interstitial lung disease in two brothers with novel compound heterozygous ABCA3 mutations. (2013)
  • Author(s): Kitazawa H, Moriya K, Niizuma H, Kawano K, Saito-Nanjo Y, Uchiyama T, Rikiishi T, Sasahara Y, Sakamoto O, Setoguchi Y, Kure S.
  • Journal Title: Eur. J. Pediatr. Volume: 172(7) Pages: 953-957
  • Peer Reviewed
• [Journal Article] T cell receptor ligation causes Wiskott-Aldrich syndrome protein degradation and F-actin assembly downregulation. (2013)
  • Author(s): Watanabe Y, Sasahara Y, Ramesh N, Massaad MJ, Looi CY, Kumaki S, Kure S, Geha RS, Tsuchiya S.
  • Journal Title: J. Allergy Clin. Immunol. Volume: 132 Pages: 648-655
  • Peer Reviewed
• [Journal Article] PBSCT is associated with poorer survival and increased chronic GVHD than BMT in Japanese patients with acute leukemia and HLA-matched sibling donor. (2013)
  • Author(s): Shinzato A, Tabuchi K, Atsuta Y, Inoue M, Inagaki J, Yabe H, Koh K, Kato K, Ohta H, Kigasawa H, Kitoh T, Ogawa A, Takahashi Y, Sasahara Y, Kato S, Adachi S.
  • Journal Title: Pediatr. Blood & Cancer Volume: 60(9) Pages: 1573-1579
  • Peer Reviewed
• [Journal Article] Successful treatment with rituximab of an infant with refractory autoimmune hemolytic anemia. (2013)
  • Author(s): Moriya K, Matsuhashi T, Onuma M, Niizuma H, Rikiishi T, Asada H, Suzuki J, Sasahara Y, Kure S.
  • Journal Title: Int. J. Hematol. Volume: 98(2) Pages: 237-239
  • NAID: 10031194740
  • Peer Reviewed
• [Journal Article] A case series of CAEBV of children and young adults treated with reduced intensity conditioning and allogeneic bone marrow transplantation; a single center study. (2013)
  • Author(s): Watanabe Y, Sasahara Y, Satoh M, Looi CY, Katayama S, Suzuki T, Suzuki N, Ouchi M, Horino S, Moriya K, Nanjyo Y, Onuma M, Kitazawa H, Irie M, Niizuma H, Uchiyama T, Rikiishi T, Kumaki S, Minegishi M, Wada T, Yachie A, Tsuchiya T, Kure S.
  • Journal Title: Eur. J. Haematol. Volume: 91 Pages: 242-248
  • Peer Reviewed
• [Journal Article] Gene therapy model of X-linked severe combined immunodeficiency using a modified foamy virus vector. (2013)
  • Author(s): Horino S, Uchiyama T, So T, Nagashima H, Sun S, Sato M, Asao A, Haji Y,
  • Journal Title: PLos One Volume: 8(8)
  • Peer Reviewed
• [Journal Article] B-cell function after unrelated umbilical cord blood transplantation using a minimal-intensity conditioning regimen in patients with X-SCID. (2013)
  • Author(s): Kumaki S, Sasahara Y, Kamachi Y, Muramatsu H, Morio T, Goi K, Sugita K, Urabe T, Takada H, Kojima S, Tsuchiya S, Hara T.
  • Journal Title: Int. J. Hematol. Volume: 98 Pages: 355-360
  • Peer Reviewed
• [Journal Article] 総説 小型および正常サイズの血小板を有する先天性血小板減少症の診断と分子病態における最近の知見 (2013)
  • Author(s): 笹原洋二、大内芽里、今泉益栄
  • Journal Title: 日本小児血液がん学会雑誌 Volume: 50(2) Pages: 186-191
  • Peer Reviewed
• [Journal Article] 免疫不全症を伴う特徴的な症候群 その１―Wiskott-Aldrich症候群、高IgE症候群― (2013)
  • Author(s): 笹原洋二
  • Journal Title: 小児科診療 Volume: 76(3) Pages: 407-412
• [Journal Article] Wiskott-Aldrich syndrome presenting with a clinical picture mimicking juvenile myelomonocytic leukemia. (2013)
  • Author(s): Yoshimi A, Kamachi Y, Imai K, Watanabe N, Kobayashi R, Yoshida M, Kobayashi C, Hama A, Muramatsu H, Sasahara Y, Morio T, Nonoyama S, Ehl S, Manabe A, Niemeyer C, Kojima S, et al.
  • Journal Title: Pediatr Blood & Cancer Volume: 60 Pages: 836-841
  • Peer Reviewed
• [Journal Article] Long-term outcome of childhood aplastic anemia patients who underwent allogeneic hematopoietic stem cell transplantation from an HLA-matched sibling donor in Japan. (2013)
  • Author(s): Kikuchi A, Yabe H, Kato K, Koh K, Inagaki J, Sasahara Y, Suzuki R, YoshidaN, Kudo K, Kobayashi R, Tabuchi K, Kawa K, Kojima S, for the Japanese Hematopoietic Cell Transplantation Registry
  • Journal Title: Bone Marrow Transplant Volume: in press
  • Peer Reviewed
• [Journal Article] Interstitial lung disease in two brothers with novel compound heterozygous ABCA3 mutations. (2013)
  • Author(s): Kitazawa H, Moriya K, Niizuma H, Kawano K, Saito-Nanjo Y, Uchiyama T, Rikiishi T, Sasahara Y, Sakamoto O, Setoguchi Y, Kure S.
  • Journal Title: Eur J Pediatr Volume: in press
  • Peer Reviewed
• [Journal Article] Degradation of the Wiskott-Aldrich syndrome protein by calpain and the ubiquitin-proteasome pathway regulates T cell receptor (TCR) driven F-actin assembly. (2013)
  • Author(s): Watanabe Y, Sasahara Y, Ramesh N, Massaad MJ, Looi CY, Kumaki S, Kure S, Geha RS, Tsuchiya S.
  • Journal Title: J Allergy Clin Immunol Volume: in press
  • Peer Reviewed
• [Journal Article] 小型および正常サイズの血小板を有する先天性血小板減少症の診断と分子病態における最近の知見 (2013)
  • Author(s): 笹原洋二
  • Journal Title: 日本小児血液がん学会雑誌 Volume: in press
  • Peer Reviewed
• [Journal Article] Second allogeneic hematopoietic SCT for relapsed ALL in children. (2012)
  • Author(s): Kato M , Horikoshi Y , Okamoto Y, Takahashi Y, Hasegawa D, Koh K, Takita J, Inoue M, Kigasawa H, Ogawa A, Sasahara Y, Kawa K, Yabe H, Sakamaki H, Suzuki R, Kato K.
  • Journal Title: Bone Marrow Transplant Volume: 1 Pages: 1-5
  • Peer Reviewed
• [Journal Article] Casitas B-cell lymphoma mutation in childhood T-cell acute lymphoblastic leukemia. (2012)
  • Author(s): Saito Y, Aoki Y, Muramatsu H, Makishima H, Maciejewski JP, Imaizumi M, Rikiishi T, Sasahara Y, Kure S, Niihori T, Tsuchiya S, Kojima S, and Matsubara Y.
  • Journal Title: Leukemia Res Volume: 36 Pages: 1009-1015
  • Peer Reviewed
• [Journal Article] Development of a multi-step leukemogenesis model of MLL-rearranged leukemia using humanized mice. (2012)
  • Author(s): Moriya K, Suzuki M, Watanabe Y, Takahashi T, Aoki Y, Uchiyama T, Kumaki S, Sasahara Y, Minegishi M, Kure S, Tsuchiya S, Sugamura K, Ishii N.
  • Journal Title: PLoS One Volume: 7
  • Peer Reviewed
• [Journal Article] Stem cell transplantation for pediatric patients with non-anaplastic peripheral T-cell lymphoma in Japan. (2012)
  • Author(s): Kobayashi R, Fujita N, Mitsui T, Iwasaki F, Suzumiya J, Kuroda H, Nishimura R,Sasahara Y, Takeshita Y, Kato K, Okumura H, Sakamaki H, Yabe H, Kawa K, Kato K, Suzuki R.
  • Journal Title: Br J Hematol Volume: 159 Pages: 88-93
  • Peer Reviewed
• [Journal Article] 複合免疫不全症 (2012)
  • Author(s): 笹原洋二
  • Journal Title: 小児の発熱A to Z Volume: 増刊号 Pages: 155-159
• [Journal Article] DiGeorge症候群 (2012)
  • Author(s): 笹原洋二
  • Journal Title: 小児疾患の診断治療基準 改訂４版 Volume: 44 Pages: 234-235
• [Journal Article] Autoantibodies to villin occur frequently in IPEX, a severe immune dysregulatiosyndrome caused by mutation of FOXP3. (2011)
  • Author(s): Kobayashi I, Kubota M, Sasahara Y, Whitesell L, Ariga T, et al.
  • Journal Title: Clin. Immunol. Volume: 141 Pages: 83-89
  • Peer Reviewed
• [Journal Article] Identification of FOXP3-negative regulatory T-like (CD4+CD25+CD127low)cells in patients with immune dysregulation, polyendocrinopathy, enteropathy, X-linked syndrome. (2011)
  • Author(s): Otsubo K, Kanegane H, Sasahara Y, Ochs HD, Miyawaki T, et al.
  • Journal Title: Clin. Immunol. Volume: 141 Pages: 111-120
  • Peer Reviewed
• [Journal Article] Octa-arginine mediated delivery of wild-type Lnk inhibits Jak2-dependent leukemia cell growth by promoting apoptosis. (2011)
  • Author(s): Chung YL, Sasahara Y, Futaki S, Tsuchiya S, Kumaki S, et al.
  • Journal Title: PLOS One Volume: 6(8)
  • Peer Reviewed
• [Journal Article] Nationwide survey of patients with primary immunodeficiency diseases in Japan. (2011)
  • Author(s): Ishimura M, Takada H, Sasahara Y, Miyawaki T, Hara T, et al.
  • Journal Title: J. Clin. Immunol. Volume: 1 Pages: 1-5
  • NAID: 120005133089
  • Peer Reviewed
• [Presentation] 招待講演 ITPと鑑別が必要な先天性血小板減少症・免疫不全症 (2015)
  • Author(s): 笹原洋二
  • Organizer: 第15回若葉小児科臨床研究会
  • Place of Presentation: 生田会館（神戸）
  • Year and Date: 2015-02-21
  • Invited
• [Presentation] 教育講演 ITPと鑑別が必要な血小板疾患 (2014)
  • Author(s): 笹原洋二
  • Organizer: 第56回日本小児血液がん学会学術集会
  • Place of Presentation: 岡山コンベンションセンター（岡山）
  • Year and Date: 2014-11-28 – 2014-11-30
  • Invited
• [Presentation] Tcell receptor ligation causes Wiskott-Aldrich syndrome protein degradation and F-actin assembly downregulation in T cells. (2014)
  • Author(s): Sasahara Y, Watanabe Y, Massaad MJ, Ramesh N, Geha RS.
  • Organizer: 6th Biennial Meeting of the European Society for Immunodeficiencies.
  • Place of Presentation: プラハコングレスセンター（プラハ）
  • Year and Date: 2014-10-29 – 2014-11-01
• [Presentation] 原発性免疫不全症に対する造血幹細胞移植ー日本全体の概要と当科症例からー (2014)
  • Author(s): 笹原洋二、佐藤大記、齋藤麻耶子、森谷邦彦、渡辺祐子、小沼正栄、力石健、久間木悟、峯岸正好、土屋滋、呉繁夫
  • Organizer: 第37回仙台BMT懇話会
  • Place of Presentation: 仙台市
• [Presentation] 原発性免疫不全症から考察する小児IBD (2014)
  • Author(s): 笹原洋二
  • Organizer: 第14回日本小児IBD研究会
  • Place of Presentation: 東京都
  • Invited
• [Presentation] WASPとIL-10受容体遺伝子変異を同定した乳児期発症炎症性腸疾患2症例の臨床的検討 (2013)
  • Author(s): 笹原洋二、大内芽里、鈴木信、鈴木資、片山紗乙莉、入江正寛、呉繁夫、浅田洋司、角田文彦、虻川大樹
  • Organizer: 第116回日本小児科学会学術集会
  • Place of Presentation: 広島市
• [Presentation] シンポジウム3 小児免疫不全症の現状と展望 免疫不全症に対する造血幹細胞移植 (2013)
  • Author(s): 笹原洋二
  • Organizer: 第55回日本小児血液がん学会学術集会
  • Place of Presentation: 福岡市
  • Invited
• [Presentation] 当科におけるCAEBV移植5症例の検討 (2013)
  • Author(s): 笹原洋二 大内芽里 鈴木信 堀野智史 小沼正栄 入江正寛 内山徹 力石健 峯岸正好 呉繁夫
  • Organizer: 第36回仙台BMT懇話会
  • Place of Presentation: 仙台市
• [Presentation] WASPとIL-10受容体遺伝子変異を同定した乳児期発症炎症性腸疾患の2症例 (2013)
  • Author(s): 笹原洋二、鈴木資、片山紗乙莉、大内芽里、鈴木信、小沼正栄、内山徹、入江正寛、力石健、呉繁夫
  • Organizer: 第3回東北小児感染免疫研究会
  • Place of Presentation: 仙台市
• [Presentation] 同胞間同種骨髄移植後、ドナーT細胞にEBVゲノムが確認されたCAEBVの１例 (2013)
  • Author(s): 笹原洋二、渡辺祐子、小沼正栄、入江正寛、内山徹、力石健、呉繁夫
  • Organizer: 第22回EBV感染症研究会
  • Place of Presentation: 東京都
• [Presentation] WASPとIL-10受容体遺伝子変異を同定した乳児期発症炎症性腸疾患2症例の臨床的検討 (2013)
  • Author(s): 笹原洋二、鈴木資、片山紗乙莉、大内芽里、鈴木信、小沼正栄、内山徹、入江正寛、力石健、呉繁夫
  • Organizer: 第32回東北免疫研究会
  • Place of Presentation: 仙台市
• [Presentation] Spontaneous reversionを伴う原発性免疫不全症2症例における長期的臨床経過の検討 (2012)
  • Author(s): 笹原洋二、佐藤美季、斉藤由佳、小沼正栄、入江正寛、内山徹、力石健、呉繁夫
  • Organizer: 第115回日本小児科学会学術集会
  • Place of Presentation: 福岡市
• [Presentation] RISTを施行した当科年長児および成人期WAS/XLT3症例の臨床的検討 (2012)
  • Author(s): 笹原洋二、鈴木信、大内芽里、渡辺祐子、小沼正栄、入江正寛、内山徹、力石健、峯岸正好
  • Organizer: 第25回東北BMT研究会
  • Place of Presentation: 仙台市
• [Presentation] 細胞外寄生菌に対する感染防御機構 (2012)
  • Author(s): 笹原洋二
  • Organizer: 第44回日本小児感染症学会
  • Place of Presentation: 北九州市
  • Invited
• [Presentation] Functional roles of WIP in Wiskott-Aldrich syndrome (2012)
  • Author(s): Yoji Sasahara, Yuko Watanabe, Looi Chung Yeng, Shigeru Tsuchiya, Shigeo Kure.
  • Organizer: 8th Congress of Asian Society for Pediatric Research
  • Place of Presentation: Seoul, Korea
• [Presentation] Mechanisms of ubiquitination and degradation of Wiskott-Aldrich syndrome protein inT cells. (2011)
  • Author(s): Yoji Sasahara, Yuko Watanabe, Looi Chung Yeng, Shigeru Tsuchiya.
  • Organizer: 7th Asian Society for Pediatric Research/Pediatric Academic Society 2011 joint meeting
  • Place of Presentation: Denver, CO, USA.
• [Presentation] RISTを施行した当科年長児および成人期WAS/XLT3症例の臨床的検討
  • Author(s): 笹原洋二、力石健、北沢博、内山徹、峯岸正好、久間木悟、土屋滋
  • Organizer: 第5回日本免疫不全症研究会
  • Place of Presentation: 東京