Analysis for the function of the candidate gene of Sick House Syndrome and development of disease model mouse

• Project/Area Number: 24651064
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Risk sciences of radiation/Chemicals
• Research Institution: Tokai University
• Principal Investigator: KIMURA Minoru 東海大学, 医学部, 教授 (10146706)
• Co-Investigator(Kenkyū-buntansha): KAJIWARA Kagemasa 東海大学, 医学部, 講師 (00204397) ; SAKABE Koh 東海大学, 医学部, 教授 (70162302) ; OHTSUKA Masato 東海大学, 医学部, 准教授 (90372945)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000); Fiscal Year 2014: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2013: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2012: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: シックハウス症候群 / 疾患感受性遺伝子 / 疾患モデルマウス / 遺伝子操作マウス / PNPLA6 / 有機リン / 遺伝子発現 / PNPLA6 gene / esterase / ROSA26 / CRISPR / トランスジェニックマウス / 遺伝子改変 / ゲノム編集 / 神経障害エステラーゼ / モデルマウス / OPIDN / 化学物質過敏症

Outline of Final Research Achievements

Sick building syndrome (SBS) is a set of several clinically recognizable symptoms reported by occupants of a building without a clear cause. Neuropathy target esterase (NTE) is a membrane bound serine esterase and its reaction with organophosphates (OPs) can lead to OP-induced delayed neuropathy (OPIDN) and nerve axon degeneration. We found that the enzymatic activity of NTE was significantly higher (P<0.0005) in SBS patients compared with controls. Thus, we constructed the transgenic(TG) mice with PNPLA6 gene encoding human NTE. Eight lines were established and each mouse from 8 lines showed higher activity for NTE than that in non transgenic mouse. The activity level was different in each tissue and the activity became higher in the old TG mice than that in the young TG mice. To elucidate the effect of OP for these mice, the experiments are now on going and the preliminary data showed harmful effects of OP for transgenic embryos. In addition, we try to detect the NTE-OP complex.

Research Products

• [Journal Article] Neuropathy Target Esterase (神経障害標的エステラーゼ）遺伝子導入マウスの作製 (2014)
  • Author(s): 加賀谷徹、割田貴之、三浦浩美、大塚正人、大久保朋一、坂部貢、木村穰
  • Journal Title: 臨床環境医学 Volume: 23(1)
  • Peer Reviewed / Open Access
• [Journal Article] Qualitative measurement of pain by analyzing the salivary alpha amylase. (2014)
  • Author(s): Tsuchiya, K., Saidin, MYB., Inoue, T., Kajiwara, K and Kimura, M.
  • Journal Title: Precision Engineering Volume: 38 Pages: 257-260
  • Peer Reviewed
• [Journal Article] Interaction between carbon nanotubes and human cell. (2014)
  • Author(s): Tsuchiya, K., Iimori, S., Kajiwara, K. and Kimura, M.
  • Journal Title: Precision Engineering Volume: 38 Pages: 116-120
  • Peer Reviewed
• [Journal Article] Association of sick building syndrome with neuropathy target esterase(NTE) activity in Japanese. (2013)
  • Author(s): Matsuzaka Y
  • Journal Title: Environ Toxicol Volume: 18 Pages: 111-222
  • DOI: 10.1002/tox.21839
  • Peer Reviewed
• [Journal Article] Improved loop-mediated isothermal amplification for HLA-DRB1 genotyping using RecA and a restriction enzyme for enhanced amplification. (2013)
  • Author(s): Mitsunaga S, Shimizu S, Okudaira Y, Oka A, Tanaka M, Kimura M, Kulski JK, Inoue I, Inoko H
  • Journal Title: Immunogenetics. Volume: 65(6) Pages: 405-15
  • DOI: 10.1007/s00251-013-0690-0
  • Peer Reviewed
• [Journal Article] Improvement of pronuclear injection-based targeted transgenesis (PITT) by iCre mRNA-mediated site-specific recombination. (2013)
  • Author(s): Ohtsuka M, Miura H, Hayashi H, Nakaoka H, Kimura M, Sato M, Gurumurthy CB, Inoko H
  • Journal Title: Transgenic Res. Volume: 22 Pages: 873-875
  • DOI: 10.1007/s11248-013-9703-x
  • Peer Reviewed
• [Journal Article] X-ray microtomographic visualization of Escherichia coli by metalloprotein overexpression (2013)
  • Author(s): Mizutani, R., Taguchi, K., Ohtsuka, M., Kimura, M., Takeuchi, A., Uesugi, K. and Suzuki, Y
  • Journal Title: Journal of Synchrotron Radiation Volume: 20 Pages: 581-586
  • Peer Reviewed
• [Journal Article] Improvement of pronuclear injection-based targeted transgenesis (PITT) by iCre mRNA-mediated site-specific recombination (2013)
  • Author(s): Ohtsuka, M., Miura, H., Hayashi, H., Nakaoka, H., Kimura, M.., Sato, M. and Gurumurthy , CB
  • Journal Title: Transgenic Research Volume: 111 Pages: 111-222
  • Peer Reviewed
• [Journal Article] Fluorescent transgenic mice suitable for multi-color aggregation chimera studies. (2012)
  • Author(s): Ohtsuka M, Miura H, Gurumurthy CB, Kimura M, Inoko H, Yoshimura S, Sato M
  • Journal Title: Cell Tissue Res. Volume: 350 Pages: 251-260
  • DOI: 10.1007/s00441-012-1470-0
  • Peer Reviewed
• [Journal Article] PITT: Pronuclear Injection-Based Targeted Transgenesis, a Reliable Transgene Expression Method in Mice (2012)
  • Author(s): Ohtsuka, M., Miura, H., Sato, M., Kimura, M., Inoko H., Gurumurthy, CB.
  • Journal Title: Exp Anim. Volume: 61 Pages: 489-502
  • NAID: 10031121853
• [Presentation] EndoGalCとtargeted toxinの組み合わせを用いたCRISPR系でノックアウトされた遺伝子改変細胞濃縮法の開発 (2014)
  • Author(s): 渡部聡、桜井敬之、中村伸吾、梶原景正、木村穰、佐藤正宏
  • Organizer: 第３７回日本分子生物学会年会
  • Place of Presentation: 横浜
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] NTE(神経障害標的エステラーゼ)遺伝子発現マウスと有機リン代謝 (2014)
  • Author(s): 山村勇貴、本杉奈美、三浦浩美、大塚正人、大久保朋一、吉野美千代、笹川昇、坂部貢、木村穰
  • Organizer: 第３７回日本分子生物学会年会
  • Place of Presentation: 横浜
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] 有機リンの培養細胞に及ぼす効果について(1) (2013)
  • Author(s): 割田貴之、本杉奈美、田中正史、大久保朋一、坂部貢、木村穣
  • Organizer: 第22回日本臨床環境医学会学術集会
  • Place of Presentation: 東京（日本歯科大学東京短期大学）
• [Presentation] Neuropathy target esterase(神経障害標的エステラーゼ)遺伝子導入マウスの作成 (2013)
  • Author(s): 加賀谷徹、割田貴之、三浦浩美、大塚正人、大久保朋一、坂部貢、木村穣
  • Organizer: 第22回日本臨床環境医学会学術集会
  • Place of Presentation: 東京（日本歯科大学東京短期大学）
• [Presentation] 22q11.2欠失症候群で欠失するヒトDGCR2遺伝子のTGF-β/BMP シグナルとの相互作用 (2013)
  • Author(s): 梶原景正、渡邊聡、麥倉信一郎、木村穣
  • Organizer: 第36 回日本分子生物学会
  • Place of Presentation: 神戸
• [Presentation] マウス繊維芽細胞においてSez12はTGF-ß シグナルに関与している (2013)
  • Author(s): 麥倉信一郎、渡邊聡、梶原景正、木村穣
  • Organizer: 第36 回日本分子生物学会
  • Place of Presentation: 神戸
• [Presentation] pnpla6遺伝子と有機リン感受性ートランスジェニックマウス作製と遺伝子導入細胞の性質ー (2013)
  • Author(s): 加賀谷 徹,大塚 正人,三浦 浩美,大久保 朋一,田中 正史,割田 貴之,本杉 奈美,坂部 貢,梶原 景正,加藤 明, 吉川 枝里,木村 穣
  • Organizer: 第36 回日本分子生物学会
  • Place of Presentation: 神戸
• [Presentation] 雄性不妊を呈するヒト疾患モデルマウスでの造精機能回復の試み (2013)
  • Author(s): 本杉 奈美,亀山 洋子,木村 穣
  • Organizer: 第36 回日本分子生物学会
  • Place of Presentation: 神戸
• [Presentation] 新規レクチンをコードするSez12はT細胞増殖シグナルを調節する (2012)
  • Author(s): 麥倉信一郎、渡部聡、梶原景正、木村穣
  • Organizer: 第３５回日本分子生物学会年会
  • Place of Presentation: 福岡市
• [Presentation] 22q11.2欠失症候群で欠失するヒトDGCR2遺伝子のマウスホモログSez12遺伝子の軟骨細胞分化への影響 (2012)
  • Author(s): 梶原景正、麥倉信一郎、渡部聡、木村穣
  • Organizer: 第３５回日本分子生物学会年会
  • Place of Presentation: 福岡市
• [Book] シックハウス症候群マニュアル (2013)
  • Author(s): 木村穣
  • Publisher: 東海大学出版会