Elucidation of molecular pathomechanisms and the search for therapeutic targets of brain malformations, by employing cerebral organoids generated from patients-derived neural stem/progenitor cells

2020 Fiscal Year Final Research Report

• Project/Area Number: 18H02781
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 52050:Embryonic medicine and pediatrics-related
• Research Institution: Kyoto Prefectural University of Medicine
• Principal Investigator: Itoh Kyoko 京都府立医科大学, 医学(系)研究科(研究院), 教授 (80243301)
• Co-Investigator(Kenkyū-buntansha): 藤本 崇宏 京都府立医科大学, 医学(系)研究科(研究院), 講師 (10446114) ; 伏木 信次 京都府立医科大学, 医学(系)研究科(研究院), 特任教授 (80150572)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Keywords: 胎児医学 / 脳形成異常 / 神経幹・前駆細胞 / 脳オルガノイド / 一次繊毛 / 繊毛病

Outline of Final Research Achievements

We generated cerebral organoids from human fetus-derived neuronal stem/progenitor cells (NSPCs) which were transfected by mutated FGFR3 genes and evaluated molecular pathogenesis of abnormal brain development, however; this in vitro model has not worked well. We, then, started to analyze function and regulation of primary cilia of NSPCs, because cilia are assumed to be involved in abnormal brain development (ciliopathy). The NSPCs transfected by mutated FGFR3 showed significantly shorter in the length of cilia as compared to those transfected with normal FGFR3. In addition, LIF (Leukemia inhibitory factor) treatment induced a significant shortening of primary cilia of normal NSPCs, being regulated by JAK-STAT3 pathway. We plan to evaluate expression level of cilia-related genes and spatiotemporal STAT3 expression under LIF treatment in order to evaluate functional relationship between microtubules and STAT3 protein.

Free Research Field

発生神経生物学、神経病理学

Academic Significance and Societal Importance of the Research Achievements

変異FGFR3を導入したヒト神経幹・前駆細胞（NSPCs）から脳オルガノイドを構築し、脳形成異常の分子機序を顕在化させる研究は、in vitroヒト脳モデルとして限界があった。そこで、NSPCsの一次繊毛の制御動態（Ciliopathy）が脳形成異常の基盤にあると仮説を立て、二次元培養モデルで、変異FGFR3導入、白血病阻害因子（LIF）作用により、NSPCsの一次繊毛の有意な短縮化が生じ、さらにJAK-STAT3経路が関与することを見いだした。今後、STAT3によって発現変動が誘導される繊毛関連遺伝子群、STAT3と微小管の直接作用を解析することで、繊毛病としての脳形成異常の本態に迫る。