Gene editing of iPS cells derived from patients with congenital phagocytic disorders

• Project/Area Number: 25293231
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Pediatrics
• Research Institution: Hiroshima University
• Principal Investigator: Kobayashi Masao 広島大学, 医歯薬保健学研究院, 教授 (00162016)
• Co-Investigator(Kenkyū-buntansha): Tsumura Miyuki 広島大学, 大学院医歯薬保健学研究院, 研究員 (80646274) ; Okada Satoshi 広島大学, 大学院医歯薬保健学研究院, 講師 (80457241) ; Yamamoto Takashi 広島大学, 大学院理学研究科, 教授 (90244102) ; Nakamura Kazuhiro 広島大学, 大学院医歯薬保健学研究院, 准教授 (70363059)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥18,460,000 (Direct Cost: ¥14,200,000、Indirect Cost: ¥4,260,000); Fiscal Year 2015: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000); Fiscal Year 2014: ¥6,240,000 (Direct Cost: ¥4,800,000、Indirect Cost: ¥1,440,000); Fiscal Year 2013: ¥6,890,000 (Direct Cost: ¥5,300,000、Indirect Cost: ¥1,590,000)
• Keywords: 先天性好中球減少症 / iPS細胞 / 遺伝子編集 / 人工ヌクレアーゼ / 遺伝子修復 / TALEN

Outline of Final Research Achievements

This study demonstrated the establishment of the disease-specific iPS cells from patients with severe congenital neutropenia (SCN). Differentiation of myeloid cells derived from SCN-iPS cells was defected at the level of promyelocyte and myelocyte. This defect observed in iPS cells was similar finding shown in bone marrow of SCN patients. The mutation of ELANE identified in an SCN patient were corrected by gene-editing method using TALEN. The iPS cells edited were completely differentiated to mature neutrophils, suggesting the restoration of neutropenia in SCN patients. These results suggest that disease-specific iPS cells from patients with SCN may be useful to analyze the pathogenesis of the defect of myeloid differentiation, and that the possible restoration of ELANE mutation by gene-editing method using TALEN may result in the normal myeloid differentiation of neutrophils in iPS cells.

Research Products

• [Journal Article] Impairment of IL-17A/F immunity to Candida and IFN-γ immunity to Mycobacterium in humans with inherited RORγT deficiency. (2015)
  • Author(s): Okada S, Markle JG, Deenick EK, Federico Mele F, Averbuch D, Kobayashi M, Abel L, Cooper AM, Notarangelo LD, Boisson-Dupuis S, Puel A, Sallusto F, Bustamante J, Tangye SG, Casanova JL
  • Journal Title: Science Volume: 349 Issue: 6248 Pages: 606-613
  • DOI: 10.1126/science.aaa4282
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Mosaicism of an ELANE mutation in an asymptomatic mother in a familial case of cyclic neutropenia. (2015)
  • Author(s): Hirata O, Okada S, Ttsumura M, Karakawa S, Matsumura I, Kimura Y, Mihara T, Yasunaga S, Takihara Y, Ohara O, Kobayashi M
  • Journal Title: Journal of Clinical Immunology Volume: 2015 Issue: 5 Pages: 1-5
  • DOI: 10.1007/s10875-015-0165-1
  • Peer Reviewed / Open Access
• [Journal Article] Paroxysmal nocturnal hemoglobinuria and telomere length predicts response to immunosuppressive therapy in pediatric aplastic anemia. (2015)
  • Author(s): Narita A, Muramatsu H, Sekiya Y, Okuno Y, Sakaguchi H, Nishio N, Yoshida N, Wang X, Xu Y, Kawashima N, Doisaki S, Hama A, Takahashi Y, Kudo K, Moritake H, Kobayashi M, Kobayashi R, Ito E, Yabe H, Ohga S, Ohara A, Kojima S; Japan Childhood Aplastic Anemia Study Group.
  • Journal Title: Haematologica. Volume: 100 Issue: 12 Pages: 1546-1552
  • DOI: 10.3324/haematol.2015.132530
  • Peer Reviewed / Open Access
• [Journal Article] Monogenic mutations differentially affect the quantity and quality of T follicular helper cells in patients with human primary immunodeficiencies. (2015)
  • Author(s): Ma CS, Wong N, Tangye SG, et al.
  • Journal Title: J Allergy Clin Immunol Volume: 136 Issue: 4 Pages: 993-1006
  • DOI: 10.1016/j.jaci.2015.05.036
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] STAT3 is a critical cell intrinsic regulator of human unconventional T cell numbers and function. (2015)
  • Author(s): Wilson RP, Ives ML, Rao G, Lau A, Payne K, Kobayashi M, Holland SM, Casanova JL, Uzel G, Ma CS, Tangye SG, Deenick EK
  • Journal Title: Journal of Experimental Medicine Volume: 212 Issue: 6 Pages: 855-864
  • DOI: 10.1084/jem.20141992
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] HAX1 in induced pluripotent stem cells from a patient with severe congenital neutropenia improves defective granulopoiesis. (2014)
  • Author(s): Morishima T, Watanabe K, Niwa A, Hirai H, Saida S, Tanaka T, Kato I, Umeda K, Hiramatsu H, Saito MK, Matsubara K, Adachi S, Kobayashi M, Nakahata T, Heike T
  • Journal Title: Haematologica Volume: 99(1) Issue: 1 Pages: 19-27
  • DOI: 10.3324/haematol.2013.083873
  • Peer Reviewed / Open Access
• [Journal Article] Simple diagnosis of STAT1 gain-of-function alleles in patients with chronic mucocutaneous candidiasis. (2014)
  • Author(s): Mizoguchi Y, Tsumura M, Okada S, Hirata O, Minegishi S, Imai K, Hyakuna N, Muramatsu H, Kojima S, Ozaki Y, Imai T, Takeda S, Okazaki T, Ito T, Yasunaga S, Takihara Y, Bryant VL, Kong XF, Cypowyj S, Boisson-Dupuis S, Puel A, Casanova JL, Morio T, Kobayashi M.
  • Journal Title: J. Leukoc. Biol. Volume: 95 Issue: 4 Pages: 667-76
  • DOI: 10.1189/jlb.0513250
  • Peer Reviewed / Open Access
• [Journal Article] Detection of <I>Mucor velutinosus</I> in a Blood Culture After Autologous Peripheral Blood Stem Cell Transplantation : A Pediatric Case Report (2014)
  • Author(s): Joichi Y, Chijimatsu I, Yarita K, Kamei K, Miki M, Onodera M, Harada M, Yokozaki M, Kobayashi M, Ohge H
  • Journal Title: Medical Mycology Journal Volume: 55 Issue: 2 Pages: E43-E48
  • DOI: 10.3314/mmj.55.E43
  • NAID: 130004567865
  • ISSN: 2185-6486, 2186-165X
  • Peer Reviewed
• [Journal Article] A 2-year-old Japanese girl with TNF receptor-associated periodic syndrome: A case report of the youngest diagnosed proband in Japan. (2014)
  • Author(s): Yasumura J, Wago M, Okada S, Nishikomori R, Takei S, Kobayashi M
  • Journal Title: Modern Rheumatology Volume: 20 Pages: 1-4
  • Peer Reviewed
• [Journal Article] Wnt3a stimulates maturation of impaired neutrophils developed from severe congenital neutropenia-derived pluipotent stem cells with heterozygous ELANE mutation. (2013)
  • Author(s): Hiramoto T
  • Journal Title: Proc Natl Acad Sci USA Volume: 110 Issue: 8 Pages: 3023-3028
  • DOI: 10.1073/pnas.1217039110
  • Peer Reviewed
• [Journal Article] Heterozygosity for the Y701C STAT1 mutation in a multiplex kindred with multifocal osteomyelitis (2013)
  • Author(s): Hirata, O.
  • Journal Title: Haematologica Volume: 98 Issue: 10 Pages: 1641-1649
  • DOI: 10.3324/haematol.2013.083741
  • Peer Reviewed
• [Journal Article] Naïve and memory human B cells have distinct requirements for STAT3 activation to differentiate into antibody-secreting plasma cells. (2013)
  • Author(s): Deenick EK, Avery DT, Chan A, Berglud LJ, Ives ML, Bustamante J, Boisson-Dupuis S, Tsumura M, Kobayashi M, Arkwright PD, Averbuch D, Engelhard D, Roesler J, Peake J, Wong M, Adelstein S, Choo S, Smart JM, French MA, Fulcher DA, Cook MC, Picard C, Durandy A, Klein C, Holland SM, Uzel G, Casanova JL, Ma CS, Tangye SG
  • Journal Title: Journal of Experimental Medicine Volume: 210 Issue: 12 Pages: 2739-2753
  • DOI: 10.1084/jem.20130323
  • Peer Reviewed
• [Journal Article] Signal transducer and activator of transcription 3 (STAT3) mutations underlying autosomal dominant hyper-IgE syndrome impair human CD8(+) T-cell memory formation and function. (2013)
  • Author(s): Ives ML, Ma CS, Palendira U,Chan A, Bustamante J, Boisson-Dupuis S, Arkwright PD, Engelhard D, Averbuch D, Magdorf K, Roesler J, Peake J, Wong M, Adelstein S, Choo S, Smart JM, Frnch MA, Fulcher DA, Cook MC, Picard C, Durandy A, Tsumura M, Kobayashi M, Uzel G, Casanova JL, Tangye SG, Deenick EK
  • Journal Title: Journal of Allergy and Clinical Immunology Volume: 132 Issue: 2 Pages: 400-411
  • DOI: 10.1016/j.jaci.2013.05.029
  • Peer Reviewed
• [Journal Article] : IL-21 signalling via STAT3 primes human naïve B cells to respond to IL-2 to enhance their differentiation into plasmablasts. (2013)
  • Author(s): Berglund LJ, Ma CS, Avery DT, Moens L, Deenick EK, Bustamante J, Boisson-Dupuis S, Wong M, Adelstein S, Arkwright PD,Fulcher DA, Ziegler JB, Smart JM, Kobayashi M, Casanova JL, Cook MC, Uzel G, Tangye SG
  • Journal Title: Blood Volume: 122 Issue: 24 Pages: 3940-3950
  • DOI: 10.1182/blood-2013-06-506865
  • Peer Reviewed
• [Journal Article] Identification of the integrin B3 L718P mutation in a pedigree with autosomal dominant thrombocytopenia with anisocytosis (2012)
  • Author(s): Kobayashi Y
  • Journal Title: British Journal of Haematology Volume: 31 Issue: 4 Pages: 762-772
  • DOI: 10.1111/bjh.12160
  • Peer Reviewed
• [Presentation] MSMD Patients with IFN-γ-STAT1 Signaling Defect Present Enhanced Osteoclastogenesis and Bone Resorption. (2015)
  • Author(s): Nishimura S, Tsumura M, Hirata O, Reiko Kagawa, Mizoguchi Y, Okada S, Kobayashi M
  • Organizer: The 57th American Society of Hematology Annual Meeting and Exposition
  • Place of Presentation: Orlando, FL, USA
  • Year and Date: 2015-12-05
  • Int'l Joint Research
• [Presentation] Impairment of IL-17 immunity to Candida and IFNγ immunity to Mycobacterium in humans with bi-allelic Rorc mutations. (2015)
  • Author(s): Okada S, Markle J, Kobayashi M, Bustamante J, Casanova JL
  • Organizer: The 57th American Society of Hematology Annual Meeting and Exposition
  • Place of Presentation: Orlando, FL, USA
  • Year and Date: 2015-12-05
  • Int'l Joint Research
• [Presentation] A comparison of myelopoiesis from induced pluripotent stem cells with a mutation in ELANE between cyclic neutropenia and sever congenital neutropenia. (2015)
  • Author(s): Saito S, Nishimura S, Tsumra M, Mizoguchi Y, Sakata S, Furue A, Kobayashi M
  • Organizer: The 57th American Society of Hematology Annual Meeting and Exposition
  • Place of Presentation: Orlando, FL, USA
  • Year and Date: 2015-12-05
  • Int'l Joint Research
• [Presentation] 好中球異常症：診断と造血幹細胞移植 (2015)
  • Author(s): 小林正夫
  • Organizer: 第77回日本血液学会
  • Place of Presentation: 金沢
  • Year and Date: 2015-10-16
  • Invited
• [Presentation] Excessive Nitric Oxide production of CGD neutrophils induces the down-regulation of NOS3 and EDN1 expression in human endothelial cells. (2014)
  • Author(s): Akari N. Utsunomiya, Miyuki Tsumura, Norioki Ohno, Mizuka Miki, Hiroshi Kawaguchi, Kazuhiro Nakamura and Masao Kobayashi
  • Organizer: The 56th American Society of Hematology Annual Meeting and Exposition
  • Place of Presentation: San Francisco, CA, USA
  • Year and Date: 2014-12-06 – 2014-12-09
• [Presentation] STAT1 gain-of-function in patients with chronic mucocutaneous candidiasis can be detected by the excessive phosphorylation of STAT1 in peripheral blood monocytes. (2014)
  • Author(s): Yoko Mizoguchi, Satoshi Okada, Miyuki Tsumura, Osamu Hirata, Shizuko Minegishi, Jean-Laurent Casanova, Tomohiro Morio, Masao Kobayashi
  • Organizer: The 56th American Society of Hematology Annual Meeting and Expositio
  • Place of Presentation: San Francisco, CA, USA
  • Year and Date: 2014-12-06 – 2014-12-09
• [Presentation] Siginificant augmentation of regulatory T cells in early neonatal period. (2014)
  • Author(s): Seiich Hayakawa, Seiko Maeno, Norioki Ohno, Satoshi Okada, Yutaka Nishimura, Michiko Hayashidani, Masao Kobayashi
  • Organizer: 16th Biennial Meeting of the European Society for Immunodeficiencies
  • Place of Presentation: Prague, Czech
  • Year and Date: 2014-10-29 – 2014-11-01
• [Presentation] Early elimination of Factor VIII inhibitor by ITI with high dose immunoglobulin in hemophilia A children. (2014)
  • Author(s): Yoko Mizoguchi, Aya Furue, Ikue Chijimatsu, Mizuka Miki, Keita Tomioka, Nakao Konishi, Atsushi Ono, Hiroshi Kawaguchi, Kazuhiro Nakamura and Masao Kobayashi
  • Organizer: WORLD FEDERATION OF HEMOPHILIA, 2014 World Congress
  • Place of Presentation: Melbourne, Australia
  • Year and Date: 2014-05-11 – 2014-05-15
• [Presentation] Genetic characteristic of patients with severe congenital neutropenia in Japan. (2013)
  • Author(s): Masao Kobayashi, Yoko Mizoguchi, Shuhei Karakawa, Satoshi Okada, Hiroshi Kawaguchi, Kazuhiro Nakamura
  • Organizer: 27th International Congress of Pediatrics
  • Place of Presentation: Melbourne, Australia