Search of new biomarker of leukemia focused on lipidomics

• Project/Area Number: 24590663
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Applied pharmacology
• Research Institution: Gifu Pharmaceutical University
• Principal Investigator: NAKAMURA Mitsuhiro 岐阜薬科大学, 薬学部, 教授 (30433204)
• Co-Investigator(Kenkyū-buntansha): MURATE Takashi 名古屋大学, 医学部, 教授 (30239537) ; BANNO Yoshiko 岐阜大学, 医学系研究科, 非常勤講師 (50116852)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000); Fiscal Year 2014: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2013: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2012: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: リピドミクス / 白血病 / 質量分析装置 / LCMS / セラミド / スフィンゴリン脂質 / バイオマーカー / 抗がん剤 / LC-MS / メタボノミクス

Outline of Final Research Achievements

Ceramides are the key intermediates involved in sphingolipid metabolism. Development of sensitive and specific methods is required to quantify the intra- and extracellular ceramide levels and to understand the physiological role of ceramides.
The combination of liquid chromatography with mass spectrometry can serve as a powerful tool for analyzing complex intact ceramides. We developed a new method using mass spectrometer coupled with ultra performance liquid chromatography (UPLC) for the rapid and simultaneous quantification of ceramides such as C16-, C18-, C20-, C22-, C24-, C24:1-, and C26-ceramide. This method was successfully applied to monitor ceramides in the subcellular fraction obtained from cultured cells. In conclusion, our new method was validated for ceramide analysis, and this can be used in a wide range of clinical applications and to perform biological studies for discerning the role(s) of sphingolipid metabolites.

Research Products

• [Journal Article] Increased Acid Ceramidase Expression depends on Upregulation of Androgen-dependent Deubiquitinases, USP2, in a Human Prostate Cancer Cell Line, LNCaP. (2015)
  • Author(s): Mizutani N, Inoue M, Omori Y, Ito H, Tamiya-Koizumi K, Takagi A, Kojima T, Nakamura M, Iwaki S, Nakatochi M, Suzuki M, Nozawa Y, Murate T.
  • Journal Title: J Biochem Volume: 157
  • Peer Reviewed
• [Journal Article] Increased SPHK2 transcription of human colon cancer cells in serum-depleted culture: the involvement of CREB transcription factor. (2015)
  • Author(s): Mizutani N, Omori Y, Tanaka K, Ito H, Takagi A, Kojima T, Nakatochi M, Ogiso H, Kawamoto Y, Nakamura M, Suzuki M, Kyogashima M, Koizumi K, Nozawa Y, Murate T.
  • Journal Title: J Cell Biochem Volume: 116 Issue: 10 Pages: 2227-2238
  • DOI: 10.1002/jcb.25173
  • Peer Reviewed
• [Journal Article] Interferon-γ decreases ceramides with long-chain fatty acids: possible involvement in atopic dermatitis and psoriasis (2014)
  • Author(s): Tawada C, et al. 6名
  • Journal Title: J Invest Dermatol Volume: 134 Issue: 3 Pages: 712-718
  • DOI: 10.1038/jid.2013.364
  • Peer Reviewed
• [Journal Article] Sphingosine kinase 1 expression is downregulated during differentiation of Friend cells due to decreased c-MYB. (2013)
  • Author(s): Mizutani N, Kobayashi M, Sobue S, Ichihara M, Ito H, Tanaka K, Iwaki S,Fujii S,Ito Y, Tamiya-Koizumi K, Takagi A, Kojima T, Naoe T, Suzuki M, Nakamura M, Banno Y, Nozawa Y, Murate T.
  • Journal Title: Biochimica et Biophysica Acta -Molecular Cell Research Volume: 1833 Issue: 5 Pages: 1006-16
  • DOI: 10.1016/j.bbamcr.2013.01.001
  • Peer Reviewed
• [Journal Article] Sphingosine kinase 1 plays a role in the upregulation of CD44 expression through extracellular signal-regulated kinase signaling in human colon cancer cells (2013)
  • Author(s): Kawahara S, Otsuji Y, Nakamura M, Murakami M, Murate T, Matsunaga T, Kanoh H, Seishima M, Banno Y, Hara A
  • Journal Title: Anticancer Drugs Volume: 24(5) Issue: 5 Pages: 473-83
  • DOI: 10.1097/cad.0b013e32835f705f
  • Peer Reviewed
• [Journal Article] Role of down-regulated neutral ceramidase during all-trans retinoic acid induced neuronal differentiation in SH-SY5Y neuroblastoma cells (2012)
  • Author(s): Kouji Tanaka
  • Journal Title: J.Biochem. Volume: (印刷中) Issue: 6 Pages: 611-620
  • DOI: 10.1093/jb/mvs033
  • Peer Reviewed
• [Journal Article] Transcriptional regulation of neutral sphingomyelinase 2 in all-trans retinoic acid-treated human breast cancer cell line, MCF-7. (2012)
  • Author(s): Ito H
  • Journal Title: J Biochem. Volume: 151 Issue: 6 Pages: 599-610
  • DOI: 10.1093/jb/mvs037
  • Peer Reviewed
• [Presentation] レスベラトロールによるASMase発現増加に伴う細胞傷害性 (2014)
  • Author(s): 大森由佳里、水谷直貴、杉本元、小泉恵子、高木明、小嶋哲人、中村光浩、野澤義則、村手隆
  • Organizer: 第15回Pharmaco-Hematologyシンポジウム
  • Place of Presentation: 名古屋
  • Year and Date: 2014-05-23 – 2014-05-24
• [Presentation] 前立腺癌細胞株LNCaPの酸性セラミダーゼ発現調節機構 (2014)
  • Author(s): 水谷直貴、大森由佳里、杉本元、小泉恵子、高木明、小嶋哲人、中村光浩、野澤義則、村手隆
  • Organizer: 第15回Pharmaco-Hematologyシンポジウム
  • Place of Presentation: 名古屋
  • Year and Date: 2014-05-23 – 2014-05-24
• [Presentation] UPLC Q-Tof/MSによる新規セラミド分析系の開発 (2013)
  • Author(s): 西端友里、水谷直貴、西田弥生、梅津亮冴、加藤大和、鈴木俊之、上田夏実、中山蓉子、太和田知里、加納宏行、坂野喜子、清島真理子、村手隆、中村光浩
  • Organizer: 日本薬学会第134年会
  • Place of Presentation: 熊本
• [Presentation] Simultaneous determination of ceramides by UPLC QTof MS using Kinetex Core-Shell COlumn (2013)
  • Author(s): Nakamura M., Nishibata Y., Umetsu R., Tawada C., Kanoh H., Mizutani N., Nishida Y., Tamiya-Koizumi K., Banno Y., Seishima M., Murate T.
  • Organizer: 7th International Ceramide Conferences
  • Place of Presentation: New York
• [Presentation] Regulatory mechanism of SPHK2 expression in colon cancer cell lines (2013)
  • Author(s): Mizutani N., Inoue M., Nishida Y., Ohmori Y., Suzuki M.,Koizumi K., Takagi A., Kojima T., Iwaki S., Fujii S., Nakamura M., Banno Y., Nozawa Y., Murate T.
  • Organizer: 7th International Ceramide Conferences
  • Place of Presentation: New York
• [Presentation] The mechanism of cytotoxicity by resveratrol against human leukemia cell lines (2013)
  • Author(s): Ohmori Y., Mizutani N., Inoue M., Nishida Y., Suzuki M.,Koizumi K., Takagi A., Kojima T., Iwaki S., Fujii S., Nakamura M., Nozawa Y., Murate T.
  • Organizer: 7th International Ceramide Conferences
  • Place of Presentation: New York
• [Presentation] Decrease in ceramides with long-chain fatty acids in atopic dermatitis and psoriasis possibly by interferon-gamma (2013)
  • Author(s): Kanoh H., Tawada C., Nakamura M., Fujisawa T., Banno Y., Seishima M.
  • Organizer: 22nd Congress of the European Academy of Dermatology and Venereology
  • Place of Presentation: Istanbul
• [Presentation] ストレス刺激下でのスフィンゴシンキナーゼ2の発現調節機構 (2013)
  • Author(s): 水谷直貴、井上みなみ、西田弥生、大森由香里、鈴木元、小泉恵子、高木明、小嶋哲人、岩城壮一郎、藤井聡、中村光浩、野澤義則、村手隆
  • Organizer: 第36回日本分子生物学会年会
  • Place of Presentation: 神戸
• [Presentation] レスベラトロールによる白血病細胞株への細胞障害性機序の解析 (2013)
  • Author(s): 大森由香里、水谷直貴、井上みなみ、西田弥生、鈴木元、小泉恵子、高木明、小嶋哲人、岩城壮一郎、藤井聡、中村光浩、野澤義則、村手隆
  • Organizer: 第86回日本生化学会大会
  • Place of Presentation: 横浜
• [Presentation] 前立腺癌細胞株LNCaPの酸性セラミダーゼは脱ユビキチン化酵素USP2により発現調節される (2013)
  • Author(s): 井上みなみ、水谷直貴、西田弥生、小泉恵子、鈴木元、岩城壮一郎、藤井聡、中村光浩、野澤義則、村手隆
  • Organizer: 第86回日本生化学会大会
  • Place of Presentation: 横浜
• [Presentation] Simultaneous determination of sphingolipid metabolites by LC/MS-IT-TOF using Kinetex Core-Shell HPLC column (2012)
  • Author(s): Nakamura M, Nishibata Y, Mizutani N, Itoh H, Otsuka M, Tamiya-Koizumi K, Murate T, Banno Y.
  • Organizer: 53rd International Conference on the Bioscience of Lipids
  • Place of Presentation: Canada
• [Presentation] Ceramide profiles in stratum corneum of inflammatory skin diseases analyzed by MALDI-TOF-MS (2012)
  • Author(s): Tawada C, Kanoh H, Banno Y, Nakamura M, Seishima M.
  • Organizer: 42nd Annual ESDR Meeting
  • Place of Presentation: Italy
• [Presentation] Analysis of ceramide profiles in stratum corneum of atopic dermatitis and psoriasis by MALDI-TOF-MS (2012)
  • Author(s): Tawada C, Kanoh H, Banno Y, Nakamura M, Seishima M.
  • Organizer: 2nd Eastern Asia Dermatology Congress
  • Place of Presentation: China
• [Presentation] c-Myb is the major regulator of sphingosine kinase 1 expression of friend cells: their role in terminal cell division (2012)
  • Author(s): Mizutani N, Inoue M, Nishida Y, Koizumi K, Nakamura M, Banno Y, Nozawa Y, Murate T.
  • Organizer: 53rd International Conference on the Bioscience of Lipids
  • Place of Presentation: Canada