Exploration of the responsible gene and the pathophysiology for unidentified fever syndromes by whole genome analysis and cell-based engineering

• Project/Area Number: 24390263
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Pediatrics
• Research Institution: Kyoto University
• Principal Investigator: NISHIKOMORI Ryuta 京都大学, 医学(系)研究科(研究院), 准教授 (70359800)
• Co-Investigator(Kenkyū-buntansha): HEIKE Toshio 京都大学, 医学研究科, 教授 (90190173) ; YASUMI Takahiro 京都大学, 医学研究科, 講師 (00511891)
• Co-Investigator(Renkei-kenkyūsha): OHARA Osamu かずさDNA研究所, ヒトゲノム研究部, 部長 (20370926) ; SAITO Megumu 京都大学, iPS細胞研究所, 准教授 (90535486) ; TOGUCHIDA Junya 京都大学, 再生医科学研究所, 教授 (40273502)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥17,940,000 (Direct Cost: ¥13,800,000、Indirect Cost: ¥4,140,000); Fiscal Year 2014: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2013: ¥5,980,000 (Direct Cost: ¥4,600,000、Indirect Cost: ¥1,380,000); Fiscal Year 2012: ¥6,760,000 (Direct Cost: ¥5,200,000、Indirect Cost: ¥1,560,000)
• Keywords: 自己炎症性症候群 / 炎症 / メンデル遺伝疾患 / 全エクソーム解析 / 自然免疫 / NLRP3 / 軟骨 / 不明熱 / FilaminA異常症 / FilaminA / SOX9 / CAPS / 自己炎症性疾患 / インフラマソーム / ELISA / 自己炎症 / iPS細胞 / 遺伝子診断 / 体細胞モザイク / 軟骨分化 / 単細胞解析 / 次世代シークエンサー

Outline of Final Research Achievements

We implemented Massive Parallel Sequencing technology to reduce the costs　for diagnostic sequencing and to detect somatic mosaicism. We also demonstrated that NLRP3 mosaicism causes Muckle-Wells syndrome in a collaborative study with the Spanish group. To identify the responsible genes for the unidentified but suspected cases with inflammation, trio-based whole exome sequencing was performed. We identified three genes responsible for the inflammatory diseases, FLNA, IFIH1 and NLRC4.
For the delineation of pathophysiology of epiphyseal overgrowth in CINCA syndrome, we utilized iPS technology by differentiating patient-derived iPS cells into chondrocytes. The chondrocyte tissues obtained from the mutated iPS cells showed the larger sizes, increased SOX9 expression, and increased production of extracellular matrix. NLRP3 inflammasome is not contributing to the larger chondrocyte tissues. cAMP/PKA/CREB pathway is important for the SOX9 upregulation by NLRP3 mutants.

Research Products

• [Journal Article] Enhanced chondrogenesis of induced pluripotent stem cells from patients with neonatal-onset multisystem inflammatory disease occurs via the caspase 1-independent cAMP/protein kinase A/CREB pathway (2015)
  • Author(s): Yokoyama, K. Ikeya, M. Umeda, K. Oda, H. Nodomi, S. Nasu, A. Matsumoto, Y. Izawa, K. Horigome, K. Kusaka, T. Tanaka, T. Saito, M. K. Yasumi, T. Nishikomori, R. Ohara, O. Nakayama, N. Nakahata, T. Heike, T. Toguchida, J.
  • Journal Title: Arthritis Rheumatol Volume: 67 Issue: 1 Pages: 302-314
  • DOI: 10.1002/art.38912
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Real-time single-cell imaging of protein secretion (2014)
  • Author(s): Yamagishi, M. Suzuki, N. Izawa, K. Nakahara, A. Mizuno, J. Shoji, S. Heike, T. Harada, Y. Nishikomori, R. Ohara, O.
  • Journal Title: Sci Rep Volume: 4 Issue: 1 Pages: 4736-4736
  • DOI: 10.1038/srep04736
  • Peer Reviewed / Open Access
• [Journal Article] Aicardi-Goutières syndrome is caused by IFIH1 mutations. (2014)
  • Author(s): Oda, H., Nakagawa, K., Abe, J., Awaya, T., Funabiki, M., Hijikata, A., Nishikomori, R., Funatsuka, M., Ohshima, Y., Sugawara, Y., Yasumi, T., Kato, H., Shirai, T., Ohara, O., Fujita, T., Heike, T.
  • Journal Title: America Journal of Human Genetics Volume: 95 Issue: 1 Pages: 121-125
  • DOI: 10.1016/j.ajhg.2014.06.007
  • NAID: 120005460706
  • Peer Reviewed
• [Journal Article] Somatic NLRP3 mosaicism in Muckle-Wells syndrome. A genetic mechanism shared by different phenotypes of cryopyrin-associated periodic syndromes. (2014)
  • Author(s): Nakagawa K, Kawai T, Umebayashi H, Takei S, Kobayashi N, Yashiro M, Kubota T, Koike R, Akuta N, Shimoyama K, Iwata N, Saito MK, Ohara O, Kambe N, Yasumi T, Izawa K, Kawai T, Heike T, Yagüe J, Nishikomori R, Aróstegui JI et al.
  • Journal Title: Ann Rheum Dis. Volume: in press Issue: 3 Pages: 1-8
  • DOI: 10.1136/annrheumdis-2013-204361
  • Peer Reviewed / Open Access
• [Journal Article] Safety and efficacy of canakinumab in Japanese patients with phenotypes of cryopyrin-associated periodic syndrome as established in the first open-label, phase-3 pivotal study (24-week results) (2013)
  • Author(s): Imagawa, T. Nishikomori, R. Takada, H. Takeshita, S. Patel, N. Kim, D. Lheritier, K. Heike, T. Hara, T. Yokota, S.
  • Journal Title: Clin Exp Rheumatol Volume: 31 Pages: 302-309
  • Peer Reviewed
• [Journal Article] Induced pluripotent stem cells from CINCA syndrome patients as a model for dissecting somatic mosaicism and drug discovery (2012)
  • Author(s): Tanaka T., Takahashi K., Yamane M., Tomida S., Nakamura S., Oshima K., Niwa A., Nishikomori R., Kambe N., Hara H., Mitsuyama M., Morone N., Heuse J.E., Yamamoto T., Watanabe A., Sato-Otsubo A., Ozawa S., Asaka I., Heike T., Yamanaka S., Nak
  • Journal Title: Blood Volume: 120(6) Issue: 6 Pages: 1299-308
  • DOI: 10.1182/blood-2012-03-417881
  • Peer Reviewed
• [Journal Article] Guidelines for the genetic diagnosis of hereditary recurrent fevers. (2012)
  • Author(s): Shinar, Y., L. Obici, I. Aksenti jevich, B. Bennetts, F. Austrup, I. Ceccherini, J. M. Costa, A. De Leener, M. Gattorno, U. Kania, I. Kone-Paut, S. Lezer, A. Livneh, I. Moix, R. Nishikomori, S. Ozen, L. Phylactou, L. Risom, D. Rowczenio, T. Sarkisian, M. E. van Gi jn, M. Witsch-Baumgartner, M. Morris, H. M. Hoffman, I. Touitou, and N. European Molecular Genetics Quality
  • Journal Title: Ann Rheum Dis Volume: 71 Issue: 10 Pages: 1599-1605
  • DOI: 10.1136/annrheumdis-2011-201271
  • Peer Reviewed
• [Journal Article] Detection of Base Substitution-Type Somatic Mosaicism of the NLRP3 Gene with> 99. 9% Statistical Confidence by Massively Parallel Sequencing (2012)
  • Author(s): Izawa, K., A. Hijikata, N. Tanaka, T. Kawai, M. K. Saito, R. Goldbach-Mansky, I. Aksentijevich, T. Yasumi, T. Nakahata, T. Heike, R. Nishikomori, and O. Ohara
  • Journal Title: DNA Res Volume: 19 Issue: 2 Pages: 143-52
  • DOI: 10.1093/dnares/dsr047
  • Peer Reviewed
• [Presentation] Aicardi-Goutieres syndrome is caused by IFIH1 mutations (2014)
  • Author(s): Oda, H. Nakagawa, K. Abe, J. Awaya, T. Funabiki, M. Hijikata, A. Nishikomori, R. Funatsuka, M. Ohshima, Y. Sugawara, Y. Yasumi, T. Kato, H. Shirai, T. Ohara, O. Fujita, T. Heike, T.
  • Organizer: American Society of Human Genetics
  • Place of Presentation: San Diego, USA
  • Year and Date: 2014-10-09
• [Presentation] アナキンラをもちいて治療をおこなった高IgD症候群の2例 (2014)
  • Author(s): 下寺佐栄子、西小森隆太、吉岡耕平、河合朋樹、八角高裕、平家俊男
  • Organizer: 第24回小児リウマチ学会
  • Place of Presentation: 仙台
  • Year and Date: 2014-10-03 – 2014-10-05
• [Presentation] インフラマソーム (2014)
  • Author(s): 西小森隆太、中川権史、横山宏司、平家俊男
  • Organizer: 第43回日本臨床免疫学会
  • Place of Presentation: 東京
  • Year and Date: 2014-09-25 – 2014-09-27
• [Presentation] 次世代シークエンサーを用いた“変異陰性TRAPS”における体細胞モザイクの検索 (2014)
  • Author(s): 中川権史、西小森隆太、河合朋樹、八角高裕、平家俊男
  • Organizer: 第117回日本小児科学会
  • Place of Presentation: 名古屋
  • Year and Date: 2014-04-11 – 2014-04-13
• [Presentation] 罹患者由来iPS細胞を用いたCINCA症候群における関節病態の解明 (2014)
  • Author(s): 横山宏司、西小森隆太、納富誠司郞、田中孝之、斎藤潤、梅田雄嗣、池谷真、中畑龍俊、戸口田淳也、平家俊男
  • Organizer: 第117回日本小児科学会
  • Place of Presentation: 名古屋
  • Year and Date: 2014-04-11 – 2014-04-13
• [Presentation] エクソンスキッピングに伴い非典型的な表現型を呈したFilaminA異常症の兄弟例 (2014)
  • Author(s): 小田紘嗣、河合朋樹、中川権史、日衛嶋栄太郎、井澤和司、西小森隆太、小原收、沼部博直、平家俊男
  • Organizer: 第117回日本小児科学会
  • Place of Presentation: 名古屋
  • Year and Date: 2014-04-11 – 2014-04-13
• [Presentation] Muckle-Wells症候群におけるNLRP3体細胞モザイク変異の検討 (2014)
  • Author(s): 中川権史、西小森隆太、他11名
  • Organizer: 第37回日本小児遺伝学会
  • Place of Presentation: 名古屋
  • Year and Date: 2014-04-10
• [Presentation] 患者由来iPS細胞を用いたCINCA症候群における関節病態の分子機構の解明 (2013)
  • Author(s): 横山宏司、西小森隆太、池谷真、那須輝、田中孝之、齋藤潤、梅田雄嗣、中畑龍俊、平家俊男、戸口田淳也
  • Organizer: 第３４回日本炎症再生医学会
  • Place of Presentation: 京都
• [Presentation] Understanding the pathology of the arthropathy in chronic infantile neurological cutaneous and articular syndrome by using iPS cells technology (2013)
  • Author(s): 横山宏司、西小森隆太、池谷真、那須輝、田中孝之、齋藤潤、梅田雄嗣、中畑龍俊、平家俊男、戸口田淳也
  • Organizer: 第２８回日本整形外科学会基礎学術集会
  • Place of Presentation: 千葉
• [Presentation] Muckle-Wells症候群におけるNLRP3体細胞モザイク変異の検討 (2013)
  • Author(s): 中川権史、西小森隆太、井澤和司、河合朋樹、八角高裕、河合利尚、梅林宏明、武井修治、小林法元、小原收、Eva Gonzalez-Roca、Juan I. Arostegui、平家俊男
  • Organizer: 第23回日本小児リウマチ学会総会
  • Place of Presentation: 大宮
• [Presentation] Cryopyrin-associated periodic syndromeの治療 本邦におけるcanakinumab臨床試験及びアナキンラ疫学調査について (2012)
  • Author(s): 酉小森隆太、井澤和司、河合朋樹、八角高裕、平家俊男
  • Organizer: 第40回日本臨床免疫学会
  • Place of Presentation: 東京
  • Year and Date: 2012-09-28
• [Patent(Industrial Property Rights)] 軟骨過形成疾患の予防および治療剤ならびにそのスクリーニング方法 (2014)
  • Inventor(s): 戸口田 淳也/西小森 隆太/横山 宏司/池谷 真/平家 俊男
  • Industrial Property Rights Holder: 戸口田 淳也/西小森 隆太/横山 宏司/池谷 真/平家 俊男
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2014-227500
  • Filing Date: 2014-11-07