Novel strategy to treat intractable CNS disorders, utilizing the properties of BBB-composing cells

• Project/Area Number: 20H00529
• Research Category: Grant-in-Aid for Scientific Research (A)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Medium-sized Section 52:General internal medicine and related fields
• Research Institution: Yamaguchi University
• Principal Investigator: Kanda Takashi 山口大学, 大学院医学系研究科, 教授(特命) (40204797)
• Co-Investigator(Kenkyū-buntansha): 竹下 幸男 山口大学, 医学部, 教授(連携講座) (70749829) ; 清水 文崇 山口大学, 大学院医学系研究科, 准教授 (90535254)
• Project Period (FY): 2020-04-01 – 2025-03-31
• Project Status: Completed (Fiscal Year 2024)
• Budget Amount: ¥44,590,000 (Direct Cost: ¥34,300,000、Indirect Cost: ¥10,290,000); Fiscal Year 2024: ¥8,190,000 (Direct Cost: ¥6,300,000、Indirect Cost: ¥1,890,000); Fiscal Year 2023: ¥8,840,000 (Direct Cost: ¥6,800,000、Indirect Cost: ¥2,040,000); Fiscal Year 2022: ¥8,190,000 (Direct Cost: ¥6,300,000、Indirect Cost: ¥1,890,000); Fiscal Year 2021: ¥8,840,000 (Direct Cost: ¥6,800,000、Indirect Cost: ¥2,040,000); Fiscal Year 2020: ¥10,530,000 (Direct Cost: ¥8,100,000、Indirect Cost: ¥2,430,000)
• Keywords: 血液脳関門 / 血管内皮細胞 / アストロサイト / ペリサイト / 自己抗体

Outline of Research at the Start

難治性中枢神経疾患の新規治療法開発の実現には血液脳関門という大きな障壁がある。本研究は、研究代表者の所属する教室で樹立した３種類のヒト血液脳関門由来不死化細胞株と、同細胞を用いて作成した相互細胞接着型ヒト血液脳関門in vitroモデルを用いて、2つの戦略のもとに難治性中枢神経疾患治療法の確立を試みようというものである。

Outline of Final Research Achievements

The developed interlocking three-dimensional human blood-brain barrier (BBB) model was registered in the patent registry as Japanese Patent No. 6684413, dated April 1, 2020. Furthermore, an improved version of the interlocking 3D human BBB model was developed using a novel porous membrane with a proprietary structure. Then, two additional patent applications have been filed (2024-228988, 2024-229182).
Regarding Research Strategy, six types of neurotrophic factors were successfully identified that were released from astrocytes in the lower chamber (brain side) upon administration to the upper chamber (vascular lumen side) using this model.

Academic Significance and Societal Importance of the Research Achievements

神経疾患の治療につながる神経栄養因子は多数発見されているが、ヒトの疾患に応用する際は、血液脳関門という隘路に阻まれて脳内に到達できない。血液脳関門を破壊することなく神経疾患治療につなげようという戦略をもとに、研究戦略上必須の手段である相互接着型ヒト血液脳関門三次元モデルの開発とともに創薬スクリーニング研究を進めた。この結果、BDNFの放出作用と神経軸索伸長効果を有する6種類の脂溶性化合物が同定された。本研究は、バリアーの向こう側にある細胞機能の改変という、BBBを越える創薬研究の新たな方法論として学術的意義は高いものと考える。また、この研究で得られた新薬候補の社会的意義も高いものと考える。

Research Products

• [Int'l Joint Research] Artois University(フランス)
• [Int'l Joint Research] University of Montpellier(フランス)
• [Journal Article] Secretome of brain microvascular endothelial cells promotes endothelial barrier tightness and protects against hypoxia-induced vascular leakage. (2024)
  • Author(s): Loiola RA, Hachani J, Duban-Deweer S, Sevin E, Bugno P, Kowalska A, Rizzi E, Shimizu F, Kanda T, Mysiorek C, Mazurek M, Gosselet F
  • Journal Title: Mol Med Volume: 30 Issue: 1 Pages: 132-132
  • DOI: 10.1186/s10020-024-00897-6
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Rift Valley fever virus is able to cross the human blood-brain barrier in vitro by direct infection with no deleterious effects. (2024)
  • Author(s): Quellec J, Piro-Megy C, Cannac M, Nisole S, Marty FH, Gosselet F, Shimizu F, Kanda T, Cetre-Sossah C, Salinas S.
  • Journal Title: J Virol Volume: 98 Issue: 10 Pages: 1-22
  • DOI: 10.1128/jvi.01267-24
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Autocrine TNF-α Increases Penetration of Myelin-Associated Glycoprotein Antibodies Across the Blood-Nerve Barrier in Anti-MAG Neuropathy (2023)
  • Author(s): Sato Ryota、Shimizu Fumitaka、Kuwahara Motoi、Mizukami Yoichi、Watanabe Kenji、Maeda Toshihiko、Sano Yasuteru、Takeshita Yukio、Koga Michiaki、Kusunoki Susumu、Kanda Takashi
  • Journal Title: Neurology - Neuroimmunology Neuroinflammation Volume: 10 Issue: 3
  • DOI: 10.1212/nxi.0000000000200086
  • Peer Reviewed / Open Access
• [Journal Article] Jo-1 Antibodies From Myositis Induce Complement-Dependent Cytotoxicity and TREM-1 Upregulation in Muscle Endothelial Cells (2023)
  • Author(s): Honda Masaya、Shimizu Fumitaka、Sato Ryota、Mizukami Yoichi、Watanabe Kenji、Takeshita Yukio、Maeda Toshihiko、Koga Michiaki、Kanda Takashi
  • Journal Title: Neurology - Neuroimmunology Neuroinflammation Volume: 10 Issue: 4 Pages: 1-11
  • DOI: 10.1212/nxi.0000000000200116
  • Peer Reviewed / Open Access
• [Journal Article] Highlighting In Vitro the Role of Brain-like Endothelial Cells on the Maturation and Metabolism of Brain Pericytes by SWATH Proteomics (2023)
  • Author(s): Menaceur Camille、Hachani Johan、Dib Shiraz、Duban-Deweer Sophie、Karamanos Yannis、Shimizu Fumitaka、Kanda Takashi、Gosselet Fabien、Fenart Laurence、Saint-Pol Julien
  • Journal Title: Cells Volume: 12 Issue: 7 Pages: 1010-1010
  • DOI: 10.3390/cells12071010
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] TNFα Activates the Liver X Receptor Signaling Pathway and Promotes Cholesterol Efflux from Human Brain Pericytes Independently of ABCA1 (2023)
  • Author(s): Dib Shiraz、Loiola Rodrigo Azevedo、Sevin Emmanuel、Saint-Pol Julien、Shimizu Fumitaka、Kanda Takashi、Pahnke Jens、Gosselet Fabien
  • Journal Title: International Journal of Molecular Sciences Volume: 24 Issue: 6 Pages: 5992-5992
  • DOI: 10.3390/ijms24065992
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Pioglitazone Attenuates the Effects of Peripheral Inflammation in a Human In Vitro Blood-Brain Barrier Model (2022)
  • Author(s): da Rocha Gustavo Henrique Oliveira、Loiola Rodrigo Azevedo、de Paula-Silva Marina、Shimizu Fumitaka、Kanda Takashi、Vieira Andrea、Gosselet Fabien、Farsky Sandra Helena Poliselli
  • Journal Title: International Journal of Molecular Sciences Volume: 23 Issue: 21 Pages: 12781-12781
  • DOI: 10.3390/ijms232112781
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Exploring lipophilic compounds that induce BDNF secretion in astrocytes beyond the BBB using a new multi-cultured human in vitro BBB model (2022)
  • Author(s): Fujisawa Miwako、Takeshita Yukio、Fujikawa Susumu、Matsuo Kinya、Okamoto Masashi、Tamada Masaya、Shimizu Fumitaka、Sano Yasuteru、Koga Michiaki、Kanda Takashi
  • Journal Title: Journal of Neuroimmunology Volume: 362 Pages: 577783-577783
  • DOI: 10.1016/j.jneuroim.2021.577783
  • Peer Reviewed
• [Journal Article] Heterozygous NOTCH1 Variants Cause Immune Activation and Microangiopathy (2022)
  • Author(s): Helman Guy、Zarekiani Parand、Tromp Samantha A.M.、Andrews Ashley、Botto Lorenzo D.、Bonkowsky Joshua L.、Chassevent Anna、Giorgio Elisa、Pippucci Tommaso、Wei Shen、Smith‐Hicks Constance、Vaula Giovanna、Willemsen Mich、Schimmel Mareike、Vollert Kurt、Shimizu Fumitaka、Kanda Takashi、et al
  • Journal Title: Annals of Neurology Volume: 92 Issue: 5 Pages: 895-901
  • DOI: 10.1002/ana.26477
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Interaction of surfactant coated PLGA nanoparticles with in vitro human brain-like endothelial cells (2022)
  • Author(s): Moya Elisa L.J.、Lombardo Sonia M.、Vandenhaute Elodie、Schneider Marc、Mysiorek Caroline、Treli Akif E.、Kanda Takashi、Shimizu Fumitaka、Sano Yasuteru、Maubon Nathalie、Gosselet Fabien、Gnday-Treli Nazende、Dehouck Marie-Pierre
  • Journal Title: International Journal of Pharmaceutics Volume: 621 Pages: 121780-121780
  • DOI: 10.1016/j.ijpharm.2022.121780
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] 【BBB-単なる障壁でない】血液脳関門―研究史と新しい研究の展開 (2022)
  • Author(s): 西原秀昭，神田 隆
  • Journal Title: Clinical Neuroscience Volume: 40 Pages: 1514-1518
• [Journal Article] BBB:多発性硬化症での変化 (2022)
  • Author(s): 藤澤 美和子，神田 隆
  • Journal Title: Clinical Neuroscience Volume: 40 Pages: 1579-1581
• [Journal Article] GRP78 Antibodies Are Associated With Blood-Brain Barrier Breakdown in Anti-Myelin Oligodendrocyte Glycoprotein Antibody-Associated Disorder (2021)
  • Author(s): Shimizu F, Ogawa R, Mizukami Y, Watanabe K, Hara K, Kadono C, Takahashi T, Misu T, Takeshita Y, Sano Y, Fujisawa M, Maeda T, Nakashima I, Fujihara K, Kanda T.
  • Journal Title: Neurol Neuroimmunol Neuroinflamm Volume: 9 Issue: 1
  • DOI: 10.1212/nxi.0000000000001038
  • Peer Reviewed / Open Access
• [Journal Article] HLA genotype-clinical phenotype correlations in multiple sclerosis and neuromyelitis optica spectrum disorders based on Japan MS/NMOSD Biobank data (2021)
  • Author(s): Watanabe M. Nakamura Y. Sato S. Niino M. Fukaura H. Tanaka M. Ochi H. et al
  • Journal Title: Scientific Reports Volume: 11 Issue: 1 Pages: 607-607
  • DOI: 10.1038/s41598-020-79833-7
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Protective effect of the sphingosine-1 phosphate receptor agonist siponimod on disrupted blood brain barrier function. (2021)
  • Author(s): Spampinato SF, Merlo S, Sano Y, Kanda T, Sortino MA.
  • Journal Title: Biochem Pharmacol. Volume: 186 Pages: 114465-114465
  • DOI: 10.1016/j.bcp.2021.114465
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] New BBB Model Reveals That IL-6 Blockade Suppressed the BBB Disorder, Preventing Onset of NMOSD. (2021)
  • Author(s): Takeshita Y, Fujikawa S, Serizawa K, Fujisawa M, Matsuo K, Nemoto J, Shimizu F, Sano Y, Tomizawa-Shinohara H, Miyake S, Ransohoff RM, Kanda T.
  • Journal Title: Neurol Neuroimmunol Neuroinflamm Volume: 8 Issue: 6
  • DOI: 10.1212/nxi.0000000000001076
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Glial cell line-derived neurotrophic factor and basic fibroblast growth factor derived from skeletal muscle pericytes increase the barrier function of endothelial cells in the endomysium. (2021)
  • Author(s): Eri Ishiguchi, Yasuteru Sano, Toshihiko Maeda, Fumitaka Shimizu, Miwako Fujisawa, Masaya Honda, Yukio Takeshita, Michiaki Koga, Takashi Kanda.
  • Journal Title: Clin Exp Neuroimmunol Volume: 12 Issue: 4 Pages: 258-267
  • DOI: 10.1111/cen3.12674
  • Peer Reviewed
• [Journal Article] A Triple Culture Cell System Modeling the Human Blood-Brain Barrier. (2021)
  • Author(s): Eleonora Rizzi, Clemence Deligne, Lucie Dehouck, Roberta Bilardo, Yasutero Sano, Fumitaka Shimizu, Takashi Kanda, Marina Resmini, Fabien Gosselet, Marie-Pierre Dehouck, Caroline Mysiorek.
  • Journal Title: J Vis Exp Volume: 177 Issue: 177 Pages: 30-30
  • DOI: 10.3791/63134
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Advancing human induced pluripotent stem cell‐derived blood‐brain barrier models for studying immune cell interactions (2020)
  • Author(s): Nishihara Hideaki、Gastfriend Benjamin D.、Soldati Sasha、Perriot Sylvain、Mathias Amandine、Sano Yasuteru、Shimizu Fumitaka、Gosselet Fabien、Kanda Takashi、Palecek Sean P.、Du Pasquier Renaud、Shusta Eric V.、Engelhardt Britta
  • Journal Title: The FASEB Journal Volume: 34 Issue: 12 Pages: 16693-16715
  • DOI: 10.1096/fj.202001507rr
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] The effect of IgG from CIDP and MMN on the blood-nerve barrier (2023)
  • Author(s): Fumitaka Shimizu, Ryota Sato, Yoichi Mizukami, Kenji Watanabe, Sonoko Misawa, Naoko Matsui, Yukio Takeshita, Toshihiko Maeda, Michiaki Koga, Yuishin Izumi, Satoshi Kuwabara and Takashi Kanda
  • Organizer: 第64回日本神経学会学術集会
• [Presentation] ギラン・バレー症候群で血液神経関門破綻をきたすU1-snRNP抗体の同定． (2023)
  • Author(s): 清水文崇，古賀道明，水上洋一，渡邊健司，佐藤亮太，前田敏彦，神田 隆，中森雅之
  • Organizer: 第34回日本末梢神経学会学術集会
• [Presentation] 血管炎による末梢神経障害の診断 (2023)
  • Author(s): 神田 隆
  • Organizer: 好腎好日セミナー(Web)
  • Invited
• [Presentation] 末梢神経障害の臨床 (2023)
  • Author(s): 神田 隆
  • Organizer: 日本神経学会九州地区生涯教育講演会（Web）
  • Invited
• [Presentation] 末梢神経障害の臨床 (2023)
  • Author(s): 神田 隆
  • Organizer: 第34回日本末梢神経学会学術集会
  • Invited
• [Presentation] 全身疾患と血液脳関門 (2023)
  • Author(s): 神田 隆
  • Organizer: 第14回日本小児免疫性脳炎研究会
  • Invited
• [Presentation] ヒト血液脳関門モデルの構築と自己免疫性神経疾患 (2023)
  • Author(s): 神田 隆
  • Organizer: 第64回日本神経病理学会総会学術集会
  • Invited
• [Presentation] Human BBB in vitro model for neuroimmunological disorders. (2023)
  • Author(s): Takashi Kanda
  • Organizer: 第35回日本神経免疫学会学術集会
  • Invited
• [Presentation] MAGニューロパチー患者では，血管内皮細胞の自己分泌TNF-αがMAG抗体の微小血管透過性を亢進させている (2022)
  • Author(s): 佐藤亮太，清水文崇，桑原 基，渡邉健司，水上洋一，前田敏彦，竹下幸男，古賀道明， 楠進，神田 隆
  • Organizer: 第33回日本末梢神経学会学術研究会
• [Presentation] Effect of IgG from Guillain-Barre syndrome on the blood-nerve barrier (2022)
  • Author(s): Fumitaka Shimizu, Michiaki Koga, Ryota Sato, Yoichi Mizukami, Kenji Watanabe, Natsumi Fujii, Ken Matsukage, Yukio Takeshita, Yasuteru Sano, Toshihiko Maeda and Takashi Kanda.
  • Organizer: 第63回日本神経学会学術集会
• [Presentation] Effect of immunoglobulin G from idiopathic inflammatory myopathy patients on human muscle microvascular endothelial cell． (2022)
  • Author(s): Masaya Honda, Fumitaka Shimizu, Takashi Kanda
  • Organizer: 第63回日本神経学会学術集会
• [Presentation] Effects of sera in NMO, MOG, and MS on IgG translocation to central nervous system. (2022)
  • Author(s): Miwako Fujisawa, Yukio Takeshita, Susumu Fujikawa, Kinya Mastuo, Fumitaka Shimizu, Yasuteru Sano, Michiaki Koga, Takashi Kanda.
  • Organizer: 第63回日本神経学会学術集会
• [Presentation] Autocrine TNF-α increases penetration of MAG antibodies across the blood-nerve barrier in anti-MAG neuropathy. (2022)
  • Author(s): Ryota Sato, Fumitaka Shimizu, Motoi Kuwahara, Yoichi Mizukami, Kenji Watanabe, Toshihiko Maeda, Yasuteru Sano, Yukio Takeshita, Michiaki Koga, Susumu Kusunoki, Takashi Kanda.
  • Organizer: 第63回日本神経学会学術集会
• [Presentation] 日本神経学会賞受賞講演「脳神経内科の卒前、専門医教育への取り組み」 (2022)
  • Author(s): 神田 隆
  • Organizer: 第63回日本神経学会学術集会
  • Invited
• [Presentation] Molecular targeted therapy in autoimmune neurological disorders (2022)
  • Author(s): Takashi Kanda
  • Organizer: 第1回国際臨床免疫シンポジウム
  • Int'l Joint Research / Invited
• [Presentation] 神経疾患の分子標的薬 (2022)
  • Author(s): 神田 隆
  • Organizer: 第119 回日本内科学会総会・講演会
  • Invited
• [Presentation] Blood-brain barrier-breakdown in anti-myelin oligodendrocyte glycoprotein antibody associated disorders (2021)
  • Author(s): Shimizu F, Ogawa R, Takahashi T, Takeshita Y, Misu T, Sano Y, Maeda T, Nakashima I, Fujihara K, Kanda T
  • Organizer: 第62回日本神経学会学術大会
• [Presentation] ギラン・バレー症候群での血液神経関門破綻メカニズムの解析 (2021)
  • Author(s): 清水文崇，古賀道明，佐藤亮太，竹下幸男，佐野泰照，松影 顕，藤井菜月美，前田敏彦，神田 隆
  • Organizer: 第32回日本末梢神経学会学術集会
• [Presentation] New in vitro and in ex-vivo triple co-culture BBB models and in vivo EAE model revealed that NMO-IgG increased the intracerebral transferability of NMO-IgG and satralizumab and that IL-6 blockade suppressed the NMO-IgG-induced leukocyte transmigration and barrier dysfunction, leading to prevention of onset of NMOSD. (2021)
  • Author(s): Takeshita Y, Fujikawa S, Serizawa K, Fujisawa M, Matsuo K, Nemoto J, Shimizu F, Sano Y, Tomizawa-Shinohara H, Miyake S, Ransohoff RM, Kanda T
  • Organizer: 第62回日本神経学会学術大会
• [Presentation] 視神経脊髄炎でのIL-6を介した血液脳関門破綻の病態機序と「BBBを治療する」というIL-6受容体阻害薬の新たな作用機序 (2021)
  • Author(s): 竹下幸男, 藤川 晋, 芹澤賢一，藤澤美和子,西原秀明,松尾欣哉, 清水文崇, 古賀道明,Richard Ransohoff , 神田 隆
  • Organizer: 第33回日本神経免疫学会学術集会
• [Presentation] 多発性硬化症でのCD20抗体によるB細胞療法における血液脳関門を中心とした新しい病態機序 (2021)
  • Author(s): 竹下幸男,藤澤 美和子,松尾欣哉,福迫陽人, 藤川 晋, 玉田雅也, 岡本雅史, 西原秀明, 清水文崇, 古賀道明, 神田 隆
  • Organizer: 第33回日本神経免疫学会総会
• [Presentation] ヒトin vitro BBBモデルを用いたアストロサイトからのBDNF分泌を促進させる脂質化合物の同定 (2021)
  • Author(s): 藤澤美和子, 竹下幸男, 藤川 晋, 松尾欣哉, 清水文崇, 古賀道明, 神田 隆
  • Organizer: 第33回日本神経免疫学会学術集会
• [Presentation] Intracerebral transferability of satralizumab in neuromyelitis optica using a new multi-cultured in vitro BBB model (2021)
  • Author(s): Matsuo K, Takeshita Y, Fujikawa S, Fujisawa M, Serizawa K, Shimizu F, Sano Y, Koga M, Kanda T.
  • Organizer: 第62回日本神経学会学術集会
• [Presentation] Blood-brain barrier-activation in myelin oligodendrocyte glycoprotein antibody associated disorders (2020)
  • Author(s): Shimizu F, Ogawa R, Takahashi T, Takeshita Y, Misu T, Sano Y, Maeda T, Nakashima I, Fujihara K, Kanda T
  • Organizer: 第61回日本神経学会学術大会
• [Presentation] Exploring lipophilic compounds which induce BDNF secretion from astrocytes beyond the BBB (2020)
  • Author(s): Takeshita Y, Fujikawa S, Okino T, Fujisawa M, Shimizu F, Maeda T, Sano Y, Koga M, Kanda T
  • Organizer: 第61回日本神経学会学術大会
• [Presentation] Effects of sera in NMO and MOG on IgG translocation to central nervous system (2020)
  • Author(s): Fujisawa M, Takeshita Y, Fujikawa S, Shimizu F, Maeda T, Sano Y, Koga M, Kanda T
  • Organizer: 第61回日本神経学会学術大会
• [Book] 末梢神経障害 (2022)
  • Author(s): 神田 隆
  • Total Pages: 506
  • Publisher: 医学書院
  • ISBN: 9784260049399
• [Book] 医学生・研修医のための脳神経内科第4版 (2021)
  • Author(s): 神田 隆
  • Total Pages: 680
  • Publisher: 中外医学社