Construction of new in vitro blood-brain barrier (BBB) and blood-nerve barrier (BNB) model that incorporates multi-culturing system of BBB or BNB components.

• Project/Area Number: 16K19513
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Neurology
• Research Institution: Yamaguchi University
• Principal Investigator: TAKESHITA Yukio 山口大学, 医学部附属病院, 診療助教(4日/週) (70749829)
• Project Period (FY): 2016-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000); Fiscal Year 2017: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2016: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
• Keywords: Blood-Brain Barrier / Blood-Nerve Barrier / laminin / Neuromyelitis optica / transmigration / in vitro model / 炎症細胞浸潤 / 視神経脊髄炎 / ラミニン / Blood-brain barrier / Blood-nerve barrier / migration / 血液脳関門 / 血液神経関門

Outline of Final Research Achievements

We construct new Blood-Brain Barrier (BBB) and Blood-Nerve Barrier (BNB) model incorporating multi-cultured system of BBB or BNB specific laminin isoform and conditionally immortalized human BBB or BNB cell lines. Our models are the first in vitro BBB model to evaluate the crosstalk between BBB specific laminin and other BBB components. Next, we successfully generated the first multi-cultured BBB model that allows the transmigration of leukocytes under shear forces.
Using these moldels, we evaluated how Neuromyelitis optica (NMO)-IgG induced the leukocyte transmigration. NMO-IgG induces IL-6 in astrocytes via AQP4 and IL-6 signaling to ECs increases chemokines and drives leukocyte migration.

Research Products

• [Int'l Joint Research] Cleveland Clinic/Biogen Neuroimmunology Research/Mayo Clinic(米国)
• [Journal Article] Identification of galectin-3 as a possible antibody target for secondary progressive multiple sclerosis. (2017)
  • Author(s): H Nishihara, F Shimizu, T Kitagawa, N Yamanaka, J Akada, Y Kuramitsu, Y Sano, Y Takeshita, et al.
  • Journal Title: Mult Scler. Volume: 23 Pages: 382-394
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Immunohistochemical analysis of laminin components in blood-nerve barrier and blood-brain barrier. (2017)
  • Author(s): Y Takeshita, M Omoto, S Fujikawa, and T Kanda.
  • Journal Title: Clin Exp Neuroimmunol Volume: 8 Pages: 49-53
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Establishment of a new conditionally immortalized human skeletal muscle microvascular endothelial cell line. (2017)
  • Author(s): H Sano, Y Sano, E Ishiguchi, F shimizu, M Omoto, T Maeda, H Nishihara, Y Takeshita, et al.
  • Journal Title: J Cell Physiol Volume: 232 Pages: 12-12
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Glucose-regulated protein 78 autoantibody associates with blood-brain barrier disruption in neuromyelitis optica. (2017)
  • Author(s): F shimizu, KL Schaller , GP Owens, AC Cotleur, D Kellner, Y Takeshita, et al.
  • Journal Title: Sci Transl Med Volume: 9 Pages: 394-394
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Effects of Neuromyelitis optica (NMO)-IgG at the blood-brain barrier in vitro. (2017)
  • Author(s): Y Takeshita ,B Obermeier, AC Cotleur, SF Spampinato, F Shimizu, E Yamamoto, Y Sano, YJ Kryzer, VA Lennon, T Kanda, and R.M. Ransohoff.
  • Journal Title: Neurol Neuroimmunol Neuroinflammation Volume: 4
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Effects of neuromyelitis optica-IgG at the blood-brain barrier in vitro. (2016)
  • Author(s): Takeshita Y, Obermeier B, Cotleur AC, Spampinato SF, Shimizu F, Yamamoto E, Sano Y, Kryzer TJ, Lennon VA, Kanda T, Ransohoff RM.
  • Journal Title: Neurol Neuroimmunol Neuroinflamm. Volume: 4 Issue: 1
  • DOI: 10.1212/nxi.0000000000000311
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] 血液神経関門(BNB)におけるBNB特異的基底膜ラミニンの効果について (2017)
  • Author(s): 竹下幸男, 藤川 晋, 清水文崇, 前田敏彦, 佐野泰照, 神田 隆
  • Organizer: 第29回日本神経免疫学会総会
• [Presentation] he effect of blood-brain barrier (BBB)-specific laminins for barrier function with a new in vitro BBB model incorporating multi-culturing system of BBB components (2017)
  • Author(s): Yukio Takeshita, Susumu Fujikawa, Fumitaka Shimizu, Yasuteru Sano, Toshihiko Maeda, Takashi Kanda
  • Organizer: XXIII World Congress of Neurology
• [Presentation] Construction of a new in vitro blood-brain barrier (BBB) model that incorporates triple-culturing system of BBB components (2017)
  • Author(s): Yukio Takeshita, Yuki Tomoe, Susumu Fujikawa, Yasuteru Sano, Hideaki Nishihara, Toshihiko Maeda, Fumitaka Shimizu, Takashi Kanda
  • Organizer: 69th American Academy of Neurology
• [Presentation] Construction of a new in vitro blood-brain barrier (BBB) model that incorporates triple-culturing system of BBB components (2016)
  • Author(s): TAKESHITA Y, TOMOE Y, NISHIHARA H, MAEDA T, SANO Y, KANDA T
  • Organizer: 第57回日本神経学会
  • Place of Presentation: 神戸コンベンションセンター(兵庫県神戸市)
  • Year and Date: 2016-05-18
  • Int'l Joint Research
• [Presentation] Construction of a new In vitro blood-brain barrier (BBB) model that incorporates triple culturing system of BBB components (2016)
  • Author(s): TAKESHITA Y, TOMOE Y, NISHIHARA H, MAEDA T, SANO Y, KANDA T
  • Organizer: Neuroscience2016
  • Place of Presentation: San Diego, USA
  • Int'l Joint Research
• [Patent(Industrial Property Rights)] 血液脳関門インヴィトロモデルおよび血液脳関門インヴィトロモデルの作製方法 (2017)
  • Inventor(s): 竹下幸男、神田 隆
  • Industrial Property Rights Holder: 山口大学
  • Industrial Property Rights Type: 特許
  • Filing Date: 2017
  • Overseas
• [Funded Workshop] XXIII World Congress of Neurology (2017)
• [Funded Workshop] Neuroscience (2017)
• [Funded Workshop] American Academy of Neurology (2017)
• [Funded Workshop] Neuroscience2016 (2016)
  • Place of Presentation: San Diego, USA
  • Year and Date: 2016-11-13