Establishment of effective and reliable detection system for genomic alterations in uterine adenomyosis

• Project/Area Number: 19K07708
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 50020:Tumor diagnostics and therapeutics-related
• Research Institution: Aichi Cancer Center Research Institute (2021); National Cancer Center Japan (2019-2020)
• Principal Investigator: Inoue Satoshi 愛知県がんセンター(研究所), 腫瘍免疫応答研究分野, ユニット長 (30801930)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2021: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2020: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2019: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: 子宮腺筋症 / 子宮内膜症 / 子宮内膜 / ゲノム解析 / FFPE / LCM / 分娩 / 妊娠 / KRAS / プロゲステロン受容体 / レーザーマイクロダイセクション

Outline of Research at the Start

申請者らは、子宮腺筋症51症例の全エキソン配列解析を終了し、約40%は発がんドライバー遺伝子KRASやPIK3CAに変異を有することが世界で初めて明らかになった。しかし凍結検体に含まれる腺筋症細胞の含有率の低さ（～5％）に起因する遺伝子変異頻度の低さが課題である。今回、申請者はLCM法を用いて、凍結検体から子宮腺筋症細胞を純化する。純化した腺筋症細胞を用いたゲノム解析により、高頻度にゲノム異常を検出する。同定したゲノム異常の機能解析を、疾患モデル細胞培養系を用いて行う。本研究では、高感度ゲノム異常検出法と機能解析系を組み合わせることにより、子宮腺筋症の病因・病態を分子レベルで明らかにする。

Outline of Final Research Achievements

Uterine adenomyosis is a benign disorder that often co-occurs with endometriosis and/or leiomyoma and impairs quality-of-life. In contrast to endometriosis and leiomyoma, the genomic features of adenomyosis are unknown. Our NGS analyses of adenomyosis revealed that recurrent oncogenic KRAS mutations were detected in 37% adenomyosis. Multi-regional sequencings showed that adenomyosis is an oligoclonal disease, with some mutations also detected in adjacent normal endometrium and/or co-occurring endometriosis. KRAS mutations were significantly more frequent in adenomyosis with co-occurring endometriosis, who had been pretreated with a progestin, or low PR experssion. Our first genetic characterization of adenomyosis suggest a common molecular etiology of adenomyotic and endometriotic clones, potentially explaining the frequent co-occurrence. Our findings may point towards genetically-guided precision therapy and/or relapse risk assessment after uterine sparing surgery.

Academic Significance and Societal Importance of the Research Achievements

本研究では、子宮腺筋症のゲノム異常を世界で初めて明らかにした。その結果、子宮腺筋症という疾患は、子宮内膜症と同様に正所性子宮内膜において共通の起源となるクローンを有することが明らかとなり、両疾患が高頻度に併発するという臨床上の特徴を分子レベルで説明することが出来た。さらにKRAS変異クローンは、プロゲステロン受容体発現抑制を起こし、実臨床で用いられているプロゲスチン抵抗性を獲得している可能性を示唆した。本研究は、ゲノム解析というアプロ―チにより、子宮内膜症と腺筋症との関係を分子レベルで明らかにしただけでなく、個別化医療の対象となりうる疾患であることを明らかにした点が本研究成果の意義と思われる。

Research Products

• [Journal Article] MicroSEC; Sequence error filtering pipeline for formalin-fixed and paraffin-embedded samples. (2021)
  • Author(s): Ikegami M, Kohsaka S, Hirose T, Ueno T, Inoue S, Konomata N, Yamauchi H, Mori T, Sekine S, Inamoto Y, Yatabe Y, Kobayashi H, Tanaka S, Mano H.
  • Journal Title: Commun Biol Volume: 4 Issue: 1 Pages: 1396-1396
  • DOI: 10.1038/s42003-021-02930-4
  • Peer Reviewed / Open Access
• [Journal Article] KRAS mutations in uterine endometrium are associated with gravidity and parity. (2020)
  • Author(s): Inoue, S., Yoshida, E., Fukui, Y., Ueno, T., Kawazu, M., Takeyama, R., Ikemura, M., Osuga, Y., Terao, Y., Hirota, Y., Mano, H.
  • Journal Title: Cell Death Disease Volume: 11 Issue: 5 Pages: 347-347
  • DOI: 10.1038/s41419-020-2559-0
  • Peer Reviewed / Open Access
• [Journal Article] High-throughput functional evaluation of BRCA2 variants of unknown significance. (2020)
  • Author(s): Ikegami, M., Kohsaka, S., Ueno, T., Momozawa, Y., Inoue, S., Tamura, K., Shimomura, A., Hosoya, N., Kobayashi, H., Tanaka, S., Mano, H.
  • Journal Title: Nature Communications Volume: 11 Issue: 1 Pages: 2573-2573
  • DOI: 10.1038/s41467-020-16141-8
  • Peer Reviewed / Open Access
• [Journal Article] Uterine adenomyosis is an oligoclonal disorder associated with KRAS mutations (2019)
  • Author(s): Inoue Satoshi、Hirota Yasushi、Ueno Toshihide、十人省略、Tanaka Yosuke、Sakata Seiji、Takeuchi Kengo、Muraoka Ayako、Osuka Satoko、Saito Tsuyoshi、Oda Katsutoshi、Osuga Yutaka、Terao Yasuhisa、Kawazu Masahito、Mano Hiroyuki
  • Journal Title: Nature Communications Volume: 10 Issue: 1 Pages: 1-13
  • DOI: 10.1038/s41467-019-13708-y
  • Peer Reviewed / Open Access
• [Journal Article] Differential regulation of CpG island methylation within divergent and unidirectional promoters in colorectal cancer (2019)
  • Author(s): Namba Shinichi、Sato Kazuhito、Kojima Shinya、Ueno Toshihide、Yamamoto Yoko、Tanaka Yosuke、Inoue Satoshi、Nagae Genta、Iinuma Hisae、Hazama Shoichi、Ishihara Soichiro、Aburatani Hiroyuki、Mano Hiroyuki、Kawazu Masahito
  • Journal Title: Cancer Science Volume: 110 Issue: 3 Pages: 1096-1104
  • DOI: 10.1111/cas.13937
  • Peer Reviewed / Open Access