多発性硬化症患者iPS細胞を用いた血液脳関門破綻候補遺伝子の同定とその臨床応用

利用多发性硬化症患者 iPS 细胞鉴定血脑屏障破坏的候选基因及其临床应用

• 批准号: 22K15711
• 负责人: 西原 秀昭
• 金额: $ 2.91万
• 依托单位: Yamaguchi University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Early-Career Scientists
• 财政年份: 2022
• 资助国家: 日本
• 起止时间: 2022-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-22K15711/
• 关键词: 多発性硬化症; 血液脳関門; iPS細胞; 免疫細胞浸潤

欧州の再発寛解型多発性硬化症患者(relapsing-remitting multiple sclerosis: RRMS)4例と健常人3例から作製したiPS細胞（MS患者7株，健常人6株）を研究代表者が独自に開発した方法(Nishihara et al. FASEBJ 2020)で血液脳関門(blood-brain barrier: BBB)構成内皮細胞に分化誘導した．MS患者由来BBB構成内皮細胞では剖検脳同様にBBB破綻をin vitroに再現することができた．その原因となる分子機構を解明するためにBBB構成内皮細胞からRNAを採取し，RNA-seq解析を施行した．発現変動遺伝子を用いたreactome pathway 解析を行い，MSでのBBB破綻に関係する分子機構を検索した．健常人とMS患者由来のBBB構成内皮細胞を比較し，炎症性サイトカインの存在しない条件下で400を超える発現変動遺伝子を同定した．発現変動遺伝子を元にしたreactome pathway解析の結果と，それぞれの候補遺伝子の分子機能，生物学的プロセス，細胞内局在を検討し，BBBの発達，維持に重要とされる分子を2つ(X, Y)，過去にBBBに対する機能は報告がないものの，内皮細胞の増殖に関与する分子を1つ(Z)同定した．各候補遺伝子のノックダウン方法を確立し，候補遺伝子Zに対しては小分子を用いた阻害実験を行うことで，健常人由来BBB構成内皮細胞にMS類似のBBB破綻が再現されるかを検討した．健常人由来BBB構成内皮細胞の候補Z機能を小分子により阻害することで，tight junction構成蛋白質であるclaudin-5の発現が低下し，小分子の透過性が亢進すること，つまり既報(Nishihara et al. Brain 2022)で報告したMS同様のBBB破綻が再現できることを示した．

In Europe, 4 patients with relapsing-remitting multiple sclerosis (RRMS) and 3 healthy people were treated with iPS cells (7 strains from MS patients and 6 strains from healthy people). Research representative Nishihara alone developed this method (Nishihara et al. FASEBJ 2020) The blood-brain barrier (BBB) constitutes the induction of differentiation of endothelial cells. The endothelial cells that make up the BBB of MS patients can be analyzed and reproduced in vitro. The molecular structure of the その cause is explained and the BBB constitutes endothelial cells. RNA is collected and RNA-seq analysis is carried out. The reactionome pathway is used to analyze the reactionome pathway, and the relationship between the BBB flaw in MS and the molecular mechanism is solved. Compared with normal people and MS patients, the endothelial cells that make up the BBB are the same. Under the condition of inflammatory disease, the condition is the same as that of the 400-degree ultra-low blood pressure. Reactome The result of pathway analysis, molecular function of candidate addendum, molecular function of biology, intracellular localization of cells, BBB development, maintenance of important molecules, 2(X, Y), the function of the BBB in the past has been reported, and the proliferation of endothelial cells has been determined in the same way as the molecule (Z). Each candidate's method is established, and each candidate's method is established, and the candidate's method is used to prevent small molecules.行うことで, the endothelial cells that make up the BBB of normal people are similar to the BBB flaw in normal people. In healthy people, the BBB constitutes the candidate Z function of endothelial cells, which can be blocked by small molecules and tightened. The protein that constitutes the junction is low in claudin-5, and the permeability of small molecules is high, and the permeability of small molecules is high (Nishihara et al. Brain 2022)でreportしたMS same as BBB flaw reappearanceできることをshowした．

项目成果

Modeling Brain Vasculature Immune Interactions In Vitro

• DOI: 10.1101/cshperspect.a041185
• 发表时间: 2023-09-01
• 期刊: COLD SPRING HARBOR PERSPECTIVES IN MEDICINE
• 影响因子: 5.4
• 作者: Lyck, Ruth;Nishihara, Hideaki;Engelhardt, Britta
• 通讯作者: Engelhardt, Britta

Theodor Kocher Institute/University of Bern(スイス)

伯尔尼大学西奥多·科赫研究所（瑞士）

Intrinsic blood-brain barrier dysfunction contributes to multiple sclerosis pathogenesis.

内在的血脑屏障功能障碍导致多发性硬化症的发病机制。

• DOI: 10.1093/brain/awac019
• 发表时间: 2022
• 期刊: Brain : a journal of neurology
• 作者: Nishihara,Hideaki;Perriot,Sylvain;Gastfriend,BenjaminD;Steinfort,Marel;Cibien,Celine;Soldati,Sasha;Matsuo,Kinya;Guimbal,Sarah;Mathias,Amandine;Palecek,SeanP;Shusta,EricV;Pasquier,RenaudDu;Engelhardt,Britta
• 通讯作者: Engelhardt,Britta

Clinical features of 8 patients with multifocal motor neuropathy in the long-term follow-up

8例多灶性运动神经病患者长期随访的临床特征

• DOI: 10.5692/clinicalneurol.cn-001810
• 发表时间: 2023
• 期刊: Rinsho Shinkeigaku
• 作者: Nemoto Joe;Shimizu Fumitaka;Maeda Toshihiko;Nishihara Hideaki;Koga Michiaki;Kanda Takashi
• 通讯作者: Kanda Takashi

The Modular µSiM: A Mass Produced, Rapidly Assembled, and Reconfigurable Platform for the Study of Barrier Tissue Models In Vitro.

• DOI: 10.1002/adhm.202200804
• 发表时间: 2022-09
• 期刊: ADVANCED HEALTHCARE MATERIALS
• 影响因子: 10
• 作者: McCloskey, Molly C.;Kasap, Pelin;Ahmad, S. Danial;Su, Shiuan-Haur;Chen, Kaihua;Mansouri, Mehran;Ramesh, Natalie;Nishihara, Hideaki;Belyaev, Yury;Abhyankar, Vinay V.;Begolo, Stefano;Singer, Benjamin H.;Webb, Kevin F.;Kurabayashi, Katsuo;Flax, Jonathan;Waugh, Richard E.;Engelhardt, Britta;McGrath, James L.
• 通讯作者: McGrath, James L.