オキシトシンの血液脳関門移行に関わる分子装置の解明と愛情の構築研究

2018 Fiscal Year Research-status Report

• Project/Area Number: 18K06889
• Research Institution: Kanazawa University
• Principal Investigator: 山本 靖彦 金沢大学, 医学系, 教授 (20313637)
• Co-Investigator(Kenkyū-buntansha): 棟居 聖一 金沢大学, 医学系, 助教 (10399040) ; 原島 愛 金沢大学, 医学系, 助教 (50705522)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Keywords: オキシトシン / 血液脳関門

Outline of Annual Research Achievements

オキシトシン（OT）が社会性行動を引き起こす“愛情ホルモン”として中枢神経作用を発揮するには、末梢循環から血液脳関門（BBB）を通過し、脳内へ移行することが必須となる。しかし、これまでこのような現象の存在そのものや、分子機構については全く分かっていなかった。本研究では、OTの脳内移行とその作用発揮の分子機構の解明を目的とする。本年度は、以下のことを明らかにした。
1．OTとRAGE(receptor for advanced glycation end-products)との直接結合と細胞内シグナル伝達の有無： 合成OT（ペプチド研）とヒト・マウスの精製組み換えRAGEタンパクとの直接結合を表面プラスモン共鳴法とELISAプレートで確認した。OTはRAGE細胞外領域の免疫グロブリン様ドメインVおよびC1に結合した。しかし、OTののRAGEへの結合は、RAGE細胞内シグナル伝達を引き起こさなかった。
2．RAGEによるオキシトシンの血管側から脳側への移行をin vitroで検証： 初代培養したサル脳の毛細血管内皮細胞、ラット脳周皮細胞及びラット星状細胞を含む微小血管単位で構成されるin vitro BBB透過性アッセイキット（MBT-24H、ファーマコセル株式会社）を用いて、RAGE依存的なOTのBBB通過性が確認できた。
3．合成OTを皮下注射、静脈内投与、鼻腔投与を行ってその脳移行をマウスで確認： 各種RAGE遺伝子改変マウス（全身でRAGEが欠損するRAGE-KOマウス、血管内皮細胞RAGE過剰発現のRAGE-Tgマウス、膜型RAGEノックインマウスなど）を駆使して検証した結果、血管内皮細胞上のRAGE依存的なOT脳内移行が確認できた。また、内在性OTと区別するため[13C,15N]OTを用いても、脳脊髄液、脳内への移行が確認できた。

Current Status of Research Progress

1: Research has progressed more than it was originally planned.

Reason

オキシトシン（OT）とRAGE(receptor for advanced glycation end-products)との直接結合およびRAGE依存的なOTの脳内移行が明らかになった。血中のOTが血液脳関門（BBB）を通過し、脳内へ移行する現象の実在が初めて証明され、その分子機構が明らかになるなど先駆的な成果と言える。

Strategy for Future Research Activity

血中のオキシトシン（OT）がRAGE(receptor for advanced glycation end-products)依存的に血液脳関門（BBB）を通過し、脳内へ移行することが分かった。今後は、予定通りに現在の方針をさらに推し進め、血中におけるOT結合キャリアタンパクの同定、OTの脳内移行促進法の開発、養育行動を改善する手段を創生する研究を行っていく。

Research Products

• [Int'l Joint Research] Harvard Medical School(米国)
  • Country Name: U.S.A.
  • Counterpart Institution: Harvard Medical School
• [Int'l Joint Research] Krasnoyarsk State Medical University(ロシア連邦)
  • Country Name: RUSSIA FEDERATION
  • Counterpart Institution: Krasnoyarsk State Medical University
• [Int'l Joint Research] University of Saskatchewan(カナダ)
  • Country Name: CANADA
  • Counterpart Institution: University of Saskatchewan
• [Journal Article] Tuning inflammation: inducing ectodomain shedding of receptor for advanced glycation end-products (RAGE) and formation of soluble RAGE (2019)
  • Author(s): Kawano S, Yamamoto Y
  • Journal Title: IMARS Highlights Volume: 14(1) Pages: 18-19
• [Journal Article] Vascular RAGE transports oxytocin into the brain to elicit its maternal bonding behaviour in mice (2019)
  • Author(s): Yamamoto Y, Liang M, Munesue S, et al.
  • Journal Title: Commun Biol Volume: 2 Pages: 76
  • DOI: 10.1038/s42003-019-0325-6
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Citrullinated histone H3: early biomarker of neutrophil extracellular traps in septic liver damage (2019)
  • Author(s): Nomura K, Miyashita T, Yamamoto Y, Munesue S, Harashima A, Takayama H, Fushida S, Ohta T
  • Journal Title: J Surg Res Volume: 234 Pages: 132-138
  • DOI: 10.1016/j.jss.2018.08.014
  • Peer Reviewed
• [Journal Article] Development of a Highly Potent Analogue and a Long-Acting Analogue of Oxytocin for the Treatment of Social Impairment-Like Behaviors (2019)
  • Author(s): Ichinose W, Cherepanov SM, Shabalova AA, Yokoyama S, Yuhi T, Yamaguchi H, Watanabe A, Yamamoto Y, Okamoto H, Horike S, Terakawa J, Daikoku T, Watanabe M, Mano N, Higashida H, Shuto S
  • Journal Title: J Med Chem Volume: - Pages: in press
  • DOI: 10.1021/acs.jmedchem.8b01691
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Extracellular S100A11 plays a critical role in spread of the fibroblast population in pancreatic cancers (2019)
  • Author(s): Takamatsu H, Yamamoto K, Tomonobu N, Murata H, Inoue Y, Yamauchi A, Sumardika W, Youyi C, Kinoshita R, Yamamura M, Fujiwara H, Mitsui Y, Araki K, Futami J, Saito K, Iioka H, Ruma MW, Putranto EW, Nishibori M, Kondo E, Yamamoto Y, Toyooka S, Sakaguchi M
  • Journal Title: Oncol Res Volume: - Pages: in press
  • DOI: 10.3727/096504018X15433161908259
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] RAGE-dependent potentiation of TRPV1 currents in sensory neurons exposed to high glucose (2018)
  • Author(s): Lam D, Theaker M, Momeni Z, Yamamoto Y, Jagadeeshan S, Campanucci VA
  • Journal Title: PLoS ONE Volume: 13(2) Pages: e0193312
  • DOI: 10.1371/journal.pone.0193312
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Anti-tumor effects of a non-steroidal anti-inflammatory drug zaltoprofen on chondrosarcoma via activating peroxisome proliferator-activated receptor gamma and suppressing matrix metalloproteinase-2 expression (2018)
  • Author(s): Higuchi T, Takeuchi, A, Munesue S, Yamamoto N, Hayashi K, Kimura H, Miwa S, Inatani H, Shimozaki S, Kato T, Aoki Y, Abe K, Taniguchi Y, Aiba H, Murakami H, Harashima A, Yamamoto Y, Tsuchiya H
  • Journal Title: Cancer Med Volume: 7(5) Pages: 1944-1954
  • DOI: 10.1002/cam4.1438
  • Peer Reviewed
• [Journal Article] CD38 in the nucleus accumbens is critical in paternal behavior in mice (2018)
  • Author(s): Akther S, Zhong J, Liang M, Cherepanov SM, Lopatina O, Salmina AB, Tsuji C, Tsuji T, Yamamoto Y, Yokoyama S, Higashida H
  • Journal Title: Messenger Volume: 6 Pages: 81-84
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] TRPM2, a new player, in cyclic ADP-ribose/CD38-dependent oxytocin release in the hypothalamus (2018)
  • Author(s): Tsuji T, Liang M, Lopatina O, Yuhi T, Tsuji C, Zhong J, Akther S, Zhong J, Nishimura T, Amina S, Liu HX, Hashii M, Furuhara K, Yamamoto Y, Yokoyama S, Higashida H
  • Journal Title: Messenger Volume: 6 Pages: 71-75
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Nitration and glycation turn mature NGF into a toxic factor for motor neurons: a role for p75NTRand RAGE signaling in ALS (2018)
  • Author(s): Kim MJ, Vargas MR, Harlan BA, Killoy KM, Ball L, Comte-Walters S, Gooz M, Yamamoto Y, Beckman JS, Barbeito L, Pehar M
  • Journal Title: Antioxid Redox Signal Volume: 28(18) Pages: 1587-1602
  • DOI: 10.1089/ars.2016.6966
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] In vitro anticancer effects of a RAGE inhibitor discovered using a structure-based drug design system (2018)
  • Author(s): El-Far A, Munesue S, Harashima A, Sato A, Shindo M, Nakajima S, Inada M, Tanaka M, Takeuchi A, Tsuchiya H, Yamamoto H, Shaheen HME, El-Sayed YS, Kawano S, Tanuma S, Yamamoto Y
  • Journal Title: Oncology Lett Volume: 15(4) Pages: 4627-4634
  • DOI: 10.3892/ol.2018.7902
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Soluble thrombomodulin attenuates endothelial cell damage in hepatic sinusoidal obstruction syndrome (2018)
  • Author(s): Takada S, Miyashita T, Yamamoto Y, Kanou S, Munesue S, Ohbatake Y, Nakanuma S, Okamoto K, Sakai S, Knoshita J, Makino I, Nakamura K, Tajima H, Takamura H, Ninomiya I, Fushida S, Ohta T
  • Journal Title: In Vivo Volume: 32(6) Pages: 1409-1417
  • DOI: 10.21873/invivo.11393
  • Peer Reviewed
• [Journal Article] JNK and ATF4 as two important platforms for tumor necrosis factor-α-stimulated shedding of receptor for advanced glycation endproducts (2018)
  • Author(s): Miyoshi A, Koyama S, Sasagawa-Monden M, Kadoya M, Konishi K, Shoji T, Inaba M, Yamamoto Y, Koyama H
  • Journal Title: FASEB J Volume: 33(3) Pages: 3575-3589
  • DOI: 10.1096/fj.201701553RR
  • Peer Reviewed
• [Presentation] Analysis of paternal aggression in CD38 knockout autism model mice (2019)
  • Author(s): Shabalova AA, Liang M, Zhong J, Tsuji C, Yamamoto Y, Yokoyama S, Higashida H
  • Organizer: International Symposium on Chronic Disease and Glycation Biology
  • Int'l Joint Research
• [Presentation] Development of remarkably strong and long-active oxytocin analogues for treatment of social impairment-like behavior (2019)
  • Author(s): Cherepanov SM, Wataru I, Yuhi T, Shabalova AA, Yokoyama S, Yamamoto Y, Higashida H
  • Organizer: International Symposium on Chronic Disease and Glycation Biology
  • Int'l Joint Research
• [Presentation] Maternal behavior of CD38 KO autism model mouse dam is improved by social support (2019)
  • Author(s): Tsuji T, Yokoyama S, Yamamoto Y, Higashida H, Tsuji C
  • Organizer: International Symposium on Chronic Disease and Glycation Biology
  • Int'l Joint Research
• [Presentation] RAGE-dependent leukocyte accumulation is critical pathology of cerebral vasospasm/neuronal dysfunction after subarachnoid haemorrhage (2019)
  • Author(s): Ishii H, Aida Y, Seiichi M, Hattori T, Takarada-Iemata M, Kitao Y, Nakada M, Yasuhiko Y, Hori O
  • Organizer: International Symposium on Chronic Disease and Glycation Biology
  • Int'l Joint Research
• [Presentation] Drug library screening to identify new compounds for inducing ectodomain shedding of RAGE and formation of soluble RAGE (2019)
  • Author(s): Kawano S, Miyazawa H, Munesue S, Harashima A, Tanaka M, Thoa DTM, Leerach N, Ooshima Y, Kinoshita Y, Yamamoto H, Yamamoto Y
  • Organizer: International Symposium on Chronic Disease and Glycation Biology
  • Int'l Joint Research
• [Presentation] Effects of salicylate and pyridoxamine on diabetic nephropathy (2019)
  • Author(s): Munesue S, Abouzed TK, Harashima A, Yamamoto H, Yamamoto Y
  • Organizer: International Symposium on Chronic Disease and Glycation Biology
  • Int'l Joint Research
• [Presentation] In vitro effects of a novel RAGE inhibitor on tumor malignancy in human fibrosarcoma HT1080 cells (2019)
  • Author(s): Oshima Y, Munesue S, El-Far AM, Harashima A, Takeuchi A, Tanaka M, Kawano S, Sato S, Shindo M, Nakajima S, Inada A, Tanuma S, Yamamoto H, Yamamoto Y
  • Organizer: International Symposium on Chronic Disease and Glycation Biology
  • Int'l Joint Research
• [Presentation] Beneficial effects of zaltprofen on anti-obesity/diabetes via activation of PPARγ (2019)
  • Author(s): Kimura K, Furukawa A, Watanebe K, Munesue S, Harashima A, Leerach N, Oshima Y, Tanaka M, Kawano S, Kinoshita Y, Nagashimada M, Higuchi T, Takeuchi A, Yamamoto Y
  • Organizer: International Symposium on Chronic Disease and Glycation Biology
  • Int'l Joint Research
• [Presentation] AQP1-elicited invasion and angiogenesis is mediated by thrombospondin type 1 domain containing 7A (THSD7A) in glioblastoma multiforme (2019)
  • Author(s): Oishi M, Yamamoto Y, Munesue S, Harashima S, Nakada M, Hayashi Y
  • Organizer: International Symposium on Chronic Disease and Glycation Biology
  • Int'l Joint Research
• [Presentation] 避けられない糖化反応とその生体応答系 (2018)
  • Author(s): 山本靖彦
  • Organizer: 日本生化学会東北地方会第84回例会
  • Invited
• [Presentation] グリケーション研究の潮流「グリケーションとその生体応答系」 (2018)
  • Author(s): 山本靖彦
  • Organizer: 第91回日本生化学会大会
  • Invited
• [Presentation] 糖尿病における血管病態の共通項を巡って「グリケーションとその除去酵素による糖尿病合併症」 (2018)
  • Author(s): 山本靖彦
  • Organizer: 第61回日本糖尿病学会
  • Invited
• [Presentation] 糖化生物学の病態生理「グリケーションとその生体応答系」 (2018)
  • Author(s): 山本靖彦
  • Organizer: 第41回日本分子生物学会大会
  • Invited
• [Presentation] 糖尿病合併症としての臓器障害～克服に向けて～「グリケーションと糖尿病の合併症」 (2018)
  • Author(s): 山本靖彦
  • Organizer: 第33回日本糖尿病合併症学会
  • Invited
• [Presentation] 栄養と糖尿病合併症 (2018)
  • Author(s): 山本靖彦
  • Organizer: 2018年度認定臨床栄養医金沢研修会
  • Invited
• [Presentation] Pattern-recognition receptor RAGE in health and disease (2018)
  • Author(s): 山本靖彦
  • Organizer: 第19回東京都医学総合研究所国際シンポジウム
  • Int'l Joint Research / Invited
• [Presentation] Pattern-recognition receptor RAGE: foe or friend? (2018)
  • Author(s): 山本靖彦
  • Organizer: RUSSIA-JAPAN Medical Symposium
  • Int'l Joint Research / Invited
• [Presentation] Pattern-recognition receptor RAGE: foe or friend for life? (2018)
  • Author(s): 山本靖彦
  • Organizer: 第13回国際メイラード反応シンポジウム（ISMR13）
  • Int'l Joint Research / Invited
• [Book] べインズ・ドミニチャク 生化学 (2018)
  • Author(s): 谷口直之、岩井一宏、藤井順逸、本家孝一監訳
  • Total Pages: 692
  • Publisher: エルゼビアジャパン
• [Remarks] 金沢大学医薬保健研究域医学系血管分子生物学ホームページ
  • URL: http://biochem2.w3.kanazawa-u.ac.jp/