2020 Fiscal Year Final Research Report

Establishment of an artificial osteosarcoma stem cell line by transduction of defined factors with with iPS cell technology

Research Project

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Project/Area Number	18K09031
Research Category	Grant-in-Aid for Scientific Research (C)
Allocation Type	Multi-year Fund
Section	一般
Review Section	Basic Section 56020:Orthopedics-related
Research Institution	Kobe University
Principal Investigator	Kawamoto Teruya 神戸大学, 医学研究科, 特命講師 (30420558)
Co-Investigator(Kenkyū-buntansha)	原仁美神戸大学, 医学部附属病院, 助教 (40437489) 深瀬直政神戸大学, 医学部附属病院, 医員 (60579324) 秋末敏宏神戸大学, 保健学研究科, 教授 (90379363) 青井貴之神戸大学, 科学技術イノベーション研究科, 教授 (00546997) 川上洋平神戸大学, 医学研究科, 医学研究員 (50626570)
Project Period (FY)	2018-04-01 – 2021-03-31
Keywords	骨肉腫 / がん幹細胞 / iPS細胞
Outline of Final Research Achievements	This study aimed to generate the cancer stem cell (CSC)-like cells from human osteosarcoma cells by introducing reprogramming factors, and to reveal the obtained cell characteristics. We could obtain the osteosarcoma stem cells by transduction of OCT3/4-KLF4-SOX2 genes into a human osteosarcoma cell line MG63. mRNA expression of reported CSC markers significantly increased in the cells. In the cells, WST-8 assays revealed that significantly lower cell proliferation and higher chemo-resistance were observed. After osteogenic induction, strongly positive staining of Alizarin red S and ALP was also observed. In vivo experiments, tumors from the cells were significantly larger in size, and strong immunopositivity of Ki-67, Osteocalcin and HIF1-α was observed. We could generate the osteosarcoma stem cells by introducing reprogramming factors. This method could be useful to elucidate the molecular mechanisms and the effective therapeutic targets in human osteosarcoma.
Free Research Field	骨軟部悪性腫瘍
Academic Significance and Societal Importance of the Research Achievements	本研究は，希少かつ治療困難な骨肉腫において，iPS細胞作製に用いられる技術を応用することで骨肉腫幹細胞を得ることに成功した初めての研究である．この成功によって骨肉腫においてもがん幹細胞研究を遂行することが可能となる革新的な成果であり，学術的意義が非常に高い．また得られた細胞の特性を解析することは，骨肉腫における幹細胞の機能を解明することに繋がり，骨肉腫の新たな病態理解や治療標的分子の同定とそれらに基づく新規治療開発を行うことが期待出来る。とくに新規治療開発は限られた治療法しか存在しない骨肉腫の臨床において，非常に有用な成果であり，社会的意義も非常に高いものである．