PROTEOMICS-BASED ANALYSIS OF BLOOD-BRAIN BARRIER TRANSPORT SYSTEM

2010 Fiscal Year Final Research Report

• Project/Area Number: 18109002
• Research Category: Grant-in-Aid for Scientific Research (S)
• Allocation Type: Single-year Grants
• Research Field: Medical pharmacy
• Research Institution: Tohoku University
• Principal Investigator: TERASAKI Tetsuya 東北大学, 大学院・薬学研究科, 教授 (60155463)
• Co-Investigator(Kenkyū-buntansha): OHTSUKI Sumio 東北大学, 大学院・薬学研究科, 准教授 (60323036)
• Project Period (FY): 2006 – 2010
• Keywords: 血液脳関門 / 脳毛細血管内皮細胞 / プロテオミクス / トランスポーター / ABC transporter / 質量分析 / pharmacoproteomics / 標的絶対プロテオミクス

Research Abstract

We have developed a highly sensitive and simultaneous method to quantify membrane protein on the basis of separation and identification of protein digestsby liquid chromatography. linked tandem mass spectrometry (LC-MS/MS)with multiplexed multiple/selective reaction monitoring (MRM)mode. The most significant result is that we succeeded to developed an in silico selection criteria to find appropriate peptide fragment to be quantified which will give high intensity by LC-MS/MS analysis. By using this method, we have quantified transporter proteins in the brain capillary endothelial cells, i. e., blood-brain barrier, in mouse. Moreover, we succeeded to clarify quantitative transporter protein expression in the human blood-brain barrier. We identified 11 membrane proteins which may act as transporter in the human blood-brain barrier. These findings will give strong impact for the future blood-brain barrier research, especially for the understanding of effects on the blood-brain barrier function.

Research Products

• [Journal Article] Quantitative targeted absolute proteomics of human blood-brain barrier transporters and receptors (2011)
  • Author(s): Y. Uchida, S. Ohtsuki, Y. Katsukura, C. Ikeda, T. Suzuki, J. Kamiie, T. Terasaki
  • Journal Title: J Neurochem Volume: 117(2) Pages: 333-345
  • Peer Reviewed
• [Journal Article] Atrial natriuretic peptide is eliminated from the brain by natriuretic peptide receptor-C-mediated brain-to-blood efflux transport at the blood-brain barrier (2010)
  • Author(s): S. Ito, S. Ohtsuki, Y. Katsukura, M. Funaki, Y. Koitabashi, A. Sugino, S. Murata, T. Terasaki
  • Journal Title: J. Cereb. Blood Flow Metab. Volume: 31 Pages: 457-466
  • Peer Reviewed
• [Journal Article] Targeted Absolute Proteomicsを用いたトランスポーターの新しい研究展開 (2009)
  • Author(s): 大槻純男、上家潤一、寺崎哲也
  • Journal Title: 遺伝子医学MOOK12 Pages: 48-54
• [Journal Article] 中枢への薬物分布におけるトランスポーターの機能と役割 (2009)
  • Author(s): 大槻純男
  • Journal Title: 日本薬理学雑誌 Volume: 134 Pages: 83-86
• [Journal Article] Expression of nuclear receptor mRNA and liver X receptor-mediated regulation of ABC transporter A1 at rat blood-brain barrier (2008)
  • Author(s): S. Akanuma, S. Hori, S. Ohtsuki, M. Fujiyoshi, T. Terasaki
  • Journal Title: Neurochem. Int. Volume: 52 Pages: 669-674
  • Peer Reviewed
• [Journal Article] Quantitative atlas of membrane transporter proteins : Development and application of a highly sensitive simultaneous LC/MS/MS method combined with novel in-silico peptide selection criteria (2008)
  • Author(s): J. Kamiie, S. Ohtsuki, R. Iwase, K. Ohmine, Y. Katsukura, K. Yanai, Y. Sekine, Y. Uchida, S. Ito, T. Terasaki
  • Journal Title: Pharm. Res. Volume: 25 Pages: 1469-1483
  • Peer Reviewed
• [Journal Article] mRNA expression levels of tight junction protein genes in mouse brain capillary endothelial cells highly purified by magnetic cell sorting (2008)
  • Author(s): S. Ohtsuki, H. Yamaguchi, Y. Katsukura, T. Asashima, T. Terasaki
  • Journal Title: J. Neurochem. Volume: 104 Pages: 147-154
  • Peer Reviewed
• [Journal Article] Multichannel liquid chromatography. tandem mass spectrometry cocktail method for comprehensive substrate characterization of multidrug resistance-associated protein 4 transporter (2007)
  • Author(s): Y. Uchida, J. Kamiie, S. Ohtsuki, T. Terasaki
  • Journal Title: Pharm. Res. Volume: 24 Pages: 2281-2296
  • Peer Reviewed
• [Journal Article] Establishing a method to isolate rat brain capillary endothelial cells by magnetic cell sorting and dominant mRNA expression of multidrug resistance-associated protein 1 and 4 in highly purified rat brain capillary endothelial cells (2007)
  • Author(s): S. Ohtsuki, H. Yamaguchi, T. Asashima, T. Terasaki
  • Journal Title: Pharm. Res. Volume: 24 Pages: 588-694
  • Peer Reviewed
• [Journal Article] Contribution of carrier-mediated transport systems to the blood-brain barrier as a supporting and protecting interface for the brain (2007)
  • Author(s): S. Ohtsuki, T. Terasaki
  • Journal Title: Pharm. Res. Volume: 24 Pages: 1745-1758
  • Peer Reviewed
• [Presentation] Quantitative Targeted Absolute Proteomics of BBB Transporters : Changes with Species and Disease (2011)
  • Author(s): T. Terasaki
  • Organizer: SESSION 10 : DRUG TRANSPORTERS AND CNS PENETRATION
  • Place of Presentation: Bethesda, Maryland, USA
  • Year and Date: 2011-03-16
• [Presentation] Blood-Brain Barrier Pharmacoproteomics : Quantitative Transporter Protein Analysis for In Vitro-In Vivo, Interspecies and Diseased State Differences . Transporter Proteins III : Clinical Relevance and Applications, Session 392, Convention Center, New Orleans (2010)
  • Author(s): T. Terasaki
  • Organizer: 24th Annual Meeting of American Association of Pharmaceutical Scientists
  • Place of Presentation: New Orleans, USA
  • Year and Date: 2010-11-18
• [Presentation] Quantification of Transporter/Enzyme Protein in Human Tissue by Mass Spectrometry : A New Path to Pharmacoproteomics (PPx) (2010)
  • Author(s): T. Terasaki, S. Ohtsuki
  • Organizer: 9th International ISSX meeting, Symposium 3 : Clinical Pharmacology of Drug Transport : From Bench to Bedside
  • Place of Presentation: Istanbul, Turkey
  • Year and Date: 2010-09-05
• [Presentation] Quantitative protein analysis of human glioblastoma, Neoangiogenesis in Brain Tumours (2010)
  • Author(s): T. Terasaki, M. Nakada, C. Ikeda, Y. Hayashi, J. Hamada, Y. Sonoda, T. Kumabe, T. Tominaga, S. Ohtsuki
  • Organizer: International meeting of Signal Transduction of the Blood-Brain Barrier, Zurich 2010, Symposium 9
  • Place of Presentation: Zurich, Switzerland
  • Year and Date: 2010-09-04
• [Presentation] Development of highly sensitive simultaneous protein quantification by LC-MS/MS and its application for the research of human disease (2010)
  • Author(s): T. Terasaki, S. Ohtsuki
  • Organizer: 28th The Japan Endocrine Society (JES)Summer Seminar on Endocrinology & Metabolism (Toward the Progress and Development of Endocrinology, Session 1 : Advancement of Research Technologies)
  • Place of Presentation: Huis Ten Bosch, Nagasaki
  • Year and Date: 2010-07-08
• [Presentation] Prediction of In Vivo ADME Based on the Transporter Protein Quantification (2009)
  • Author(s): T. Terasaki
  • Organizer: 2009 AAPS Annual Meeting and Exposition
  • Place of Presentation: Los Angels, CA, USA
  • Year and Date: 20091108-12
• [Presentation] Transporter protein qunatification for the future of thransporter science. Advanced Technologies for the Future of Transporter Sciences (2009)
  • Author(s): T. Terasaki, J. Kamiie, S. Ohtsuki
  • Organizer: The 36th Congress of the International Union of Physiological Sciences (IUPS2009)
  • Place of Presentation: Kyoto, Japan
  • Year and Date: 20090728-0801
• [Presentation] Quantitative transporter protein analysis of human brain capillary endothelial (2009)
  • Author(s): T. Terasaki
  • Organizer: 8the Cerebral Vascular Biology International Conference (CVB2009)
  • Place of Presentation: Sendai
  • Year and Date: 20090628-0702
• [Presentation] New approach to drug design by quantitative membrane proteomics (2009)
  • Author(s): S. Ohtsuki
  • Organizer: Special lecture in Seoul Netional University
  • Place of Presentation: Seoul, Korea
  • Year and Date: 2009-05-20
• [Presentation] Pharmacoproteomics in Cancer Research (2008)
  • Author(s): S. Ohtsuki, R. Iwase, J. Kamiie, K. Ohmine, H. Harigae, T. Terasaki
  • Organizer: 2009 AAPS Annual Meeting and Exposition
  • Place of Presentation: Los Angels, CA, USA
  • Year and Date: 20081108-12
• [Book] トランスポーター科学最前線 (2008)
  • Author(s): 寺崎哲也
  • Total Pages: 23
  • Publisher: 京都廣川書店
• [Remarks]
  • URL: http://www.pharm.tohoku.ac.jp/~soutatsu/dds
• [Patent(Industrial Property Rights)] 質量分析計を使った膜タンパク質の定量方法 (2011)
  • Inventor(s): 上家潤一、大槻純男、寺崎哲也
  • Industrial Property Rights Holder: 国立大学法人東北大学
  • Industrial Property Number: 特許、7,901,942号
  • Filing Date: 2011-03-08
  • Overseas
• [Patent(Industrial Property Rights)] 質量分析計を使った膜タンパク質の定量方法 (2011)
  • Inventor(s): 上家潤一、大槻純男、寺崎哲也
  • Industrial Property Rights Holder: 国立大学法人東北大学
  • Industrial Property Number: 特許、4670060号
  • Filing Date: 2011-01-28
• [Patent(Industrial Property Rights)] 質量分析計を使った膜タンパク質の定量方法 (2010)
  • Inventor(s): 上家潤一、大槻純男、寺崎哲也
  • Industrial Property Rights Holder: 国立大学法人東北大学
  • Industrial Property Number: 特許、2006313253号
  • Filing Date: 2010-10-07
  • Overseas
• [Patent(Industrial Property Rights)] 質量分析計を使った膜タンパク質の定量方法 (2008)
  • Inventor(s): 上家潤一、大槻純男、寺崎哲也
  • Industrial Property Rights Holder: 国立大学法人東北大学
  • Industrial Property Number: 特許出願、06822538
  • Filing Date: 2008-06-17
  • Overseas
• [Patent(Industrial Property Rights)] 質量分析計を使った膜タンパク質の定量方法 (2008)
  • Inventor(s): 上家潤一、大槻純男、寺崎哲也
  • Industrial Property Rights Holder: 国立大学法人東北大学
  • Industrial Property Number: 特許出願、2006313253
  • Filing Date: 2008-05-22
  • Overseas
• [Patent(Industrial Property Rights)] 質量分析計を使った膜タンパク質の定量方法 (2008)
  • Inventor(s): 上家潤一、大槻純男、寺崎哲也
  • Industrial Property Rights Holder: 国立大学法人東北大学
  • Industrial Property Number: 特許出願、4197/DELNP/2008
  • Filing Date: 2008-05-16
  • Overseas
• [Patent(Industrial Property Rights)] 質量分析計を使った膜タンパク質の定量方法 (2008)
  • Inventor(s): 上家潤一、大槻純男、寺崎哲也
  • Industrial Property Rights Holder: 国立大学法人東北大学
  • Industrial Property Number: 特許出願、200680041752.6
  • Filing Date: 2008-05-08
  • Overseas
• [Patent(Industrial Property Rights)] 質量分析計を使った膜タンパク質の定量方法 (2008)
  • Inventor(s): 上家潤一、大槻純男、寺崎哲也
  • Industrial Property Rights Holder: 国立大学法人東北大学
  • Industrial Property Number: 特許出願、12/093,133
  • Filing Date: 2008-05-08
  • Overseas
• [Patent(Industrial Property Rights)] 質量分析計を使った膜タンパク質の定量方法 (2008)
  • Inventor(s): 上家潤一、大槻純男、寺崎哲也
  • Industrial Property Rights Holder: 国立大学法人東北大学
  • Industrial Property Number: 特許出願、2,628,708
  • Filing Date: 2008-05-06
  • Overseas