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KAKEN_15K15610seika.pdf
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CRISPR/CAS9を用いた子宮筋腫発生モデルの開発
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CRISPR/CAS9 オ モチイタ シキュウ キンシュ ハッセイ モデル ノ カイハツ
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CRISPR/CAS9 o mochiita shikyu kinshu hassei moderu no kaihatsu
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Development of uterine leiomyoma model using CRISPR/CAS9 genome editing system
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丸山, 哲夫
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マルヤマ, テツオ
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Maruyama, Tetsuo
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慶應義塾大学・医学部・准教授
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Research team head
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科研費研究者番号 : 10209702
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内田, 浩
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ウチダ, ヒロシ
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Uchida, Hiroshi
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慶應義塾大学・医学部・講師
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Research team member
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科研費研究者番号 : 90286534
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升田, 博隆
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マスダ, ヒロタカ
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Masuda, Hirotaka
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慶應義塾大学・医学部・講師
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Research team member
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科研費研究者番号 : 80317198
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小野, 政徳
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オノ, マサノリ
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Ono, Masanori
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慶應義塾大学・医学部・共同研究員
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Research team member
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科研費研究者番号 : 70348712
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荒瀬, 透
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アラセ, トオル
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Arase, Toru
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慶應義塾大学・医学部・共同研究員
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Research team member
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科研費研究者番号 : 20348709
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高尾, 知佳
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タカオ, トモカ
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Takao, Tomoka
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慶應義塾大学・医学部・特任助教
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Research team member
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科研費研究者番号 : 40612429
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2017
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科学研究費補助金研究成果報告書
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2016
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正常子宮平滑筋組織から分離した子宮平滑筋細胞には, 分離直後は幹様細胞が多く存在するが, その後培養を続けると幹様細胞は激減した。そこで, 幹様細胞ではなく正常子宮平滑筋細胞に対して, CRISPR/CAS9システムを用いて子宮筋腫の約70％に存在するMED12変異の導入を試みたところ, MED12変異導入細胞が得られた。これを用いて子宮筋腫に特徴的なホルモン依存性の細胞増殖とコラーゲン発現について調べた。MED12変異導入細胞では野生型と比較して増殖に影響はなかったが, 変異及びホルモン依存的にコラーゲン発現の増加傾向が認められた。現在in vivo子宮筋腫モデルの作製を進めている。
Uterine myometrial cells contained a small but significant amount of stem-like cells immediately after isolation from human myometrial tissues. The number of the stem-like cells, however, decreased considerably when they are cultured in vitro. Instead of myometrial stem-like cells, we introduced MED12 mutations, which are observed in approximately 70% of uterine leiomyomas, into human myometrium cells using CRISPR/CAS9 system, and analyzed hormone-dependent cell proliferation and collagen expression. Preliminary data showed that MED12 mutation did not affect cell growth but tended to induce collagen expression. In vivo leiomyoma model system is under development.
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研究種目 : 挑戦的萌芽研究
研究期間 : 2015～2016
課題番号 : 15K15610
研究分野 : 生殖内分泌, 婦人科腫瘍
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