子宮平滑筋肉腫におけるATRX発現とその機能及び分子機構の解明

• Project/Area Number: 21K15389
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 49020:Human pathology-related
• Research Institution: Nara Medical University
• Principal Investigator: 内山 智子 奈良県立医科大学, 医学部, 助教 (80745448)
• Project Period (FY): 2021-04-01 – 2024-03-31
• Project Status: Granted (Fiscal Year 2022)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2023: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2022: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2021: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
• Keywords: leiomyosarcoma / microRNA / ATRX

Outline of Research at the Start

FFPEを用い、免疫染色にて腫瘍中のATRXタンパク発現の有無を明らかにする。また組織標本から腫瘍部分を切り出し、腫瘍中のATRX遺伝子mRNA発現についてリアルタイムRT-PCR法で確認する。In silicoにて検索し得た、ATRXと相補性を有するmiRNA候補について、同様に発現量を検討する。さらにATRX発現消失症例と、発現の保たれた症例との間で、予後を含めた臨床病理学的検討を行う。次に、テロメア特異的FISHを行い、ALTの有無について明らかにするとともにATRX発現との関連について検討する。子宮平滑筋肉腫由来細胞を用いてATRX発現に関する機能解析を行う。

Outline of Annual Research Achievements

申請書の実験計画に基づき、平滑筋肉腫細胞株（SK-UT-1, SK-LMS-1, SKN）を用いてATRXを含めたホルモン受容体や上皮間葉マーカーについて発現を検討したところERa, HER2, CDH1, Vimentinについて発現を認めた。そこで、平滑筋肉腫細胞株においていくつかの癌関連miRNAの過剰発現によるERa, HER2, CDH1, Vimentinの発現量変化について検討したところ、miR-367,675の過剰発現により上皮間葉移行が生じていることが推定された。一方で、miR-367とmiR-675は細胞増殖能への影響は乏しかったものの、遊走能、浸潤能に影響していた。そこで、これらの機能に関連する分子群を明らかにするため、標的候補分子についてIn silicoで検索し、miR-367の推定標的候補分子をいくつか抽出し、細胞株を用いてRT-PCR法にて発現を検討したところHIPK3, MYO1B, RAB23遺伝子の発現が認められた。コントロール群とmiR-367pre導入群間で発現量を検討したところmiR-367pre群ではHIPK3, MYO1Bでは全ての細胞株で、RAB23ではSK-LMS-1及びSKNで有意に発現量が低下していた。以上より、HIPK3, MYO1B, RAB23は平滑筋肉腫細胞において発現を認め、さらにmiR-367の推定標的分子であることが判明した。またSK-LMS-1,及びSKNにおいてそれぞれのsiRNAを導入したところRAB23発現抑制によりMYO1Bの発現上昇を認めた。またRAB23またはMYO1B発現抑制によりHIPK3の発現上昇を認めたことからRAB23を上流としてMYO1B, HIPK3の発現を調節している可能性が示唆された。現在はそれぞれの分子の機能が遊走能や浸潤能あるいは細胞増殖能に関連しているかを解析中である。

Current Status of Research Progress

2: Research has progressed on the whole more than it was originally planned.

Reason

論文作成中のため上記の通り報告する。

Strategy for Future Research Activity

現在は、昨年度得られたNGSを用いた解析結果について論文作成を行うと共に、細胞実験を継続して行っている。前立腺癌細胞ではmiR-367がRAB23の発現をdownregulateし、miR-367の過剰発現やRAB23のノックダウンが癌細胞の増殖・浸潤・転移を抑制するとの報告がある。機能解析のうち現在までに細胞増殖能については、MTSassayを行ったところ、miR-367 preを導入した全ての細胞株について細胞増殖能の変化は認めなかった。HIPK3，MYO1BではRAB23と類似の報告はないが、同様にそれぞれの分子について遊走能、浸潤能を中心に機能解析を行う。細胞実験については学会発表と論文作成を目標としている。

Research Products

• [Journal Article] Clinicopathological and genetic analyses of pulmonary enteric adenocarcinoma (2022)
  • Author(s): Okada Fumi、Takeda Maiko、Fujii Tomomi、Uchiyama Tomoko、Sasaki Shoh、Matsuoka Minami、Nitta Yuji、Terada Chiyoko、Maebo Katsuya、Morita Kohei、Ishida Eiwa、Sawabata Noriyoshi、Ohbayashi Chiho
  • Journal Title: Journal of Clinical Pathology Volume: - Issue: 2 Pages: 111-115
  • DOI: 10.1136/jcp-2022-208583
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• [Journal Article] Intermittent Hypoxia Increased the Expression of DBH and PNMT in Neuroblastoma Cells via MicroRNA-375-Mediated Mechanism (2022)
  • Author(s): Takasawa Shin, Shobatake Ryogo, Takeda Yoshinori, Uchiyama Tomoko, Yamauchi Akiyo, Makino Mai, Sakuramoto-Tsuchida Sumiyo, Asai Keito, Ota Hiroyo, Itaya-Hironaka Asako
  • Journal Title: International Journal of Molecular Sciences Volume: 23 Issue: 11 Pages: 5868-5868
  • DOI: 10.3390/ijms23115868
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• [Journal Article] Upregulation of IL‐8, osteonectin, and myonectin mRNAs by intermittent hypoxia via OCT1‐ and NRF2‐mediated mechanisms in skeletal muscle cells (2022)
  • Author(s): Takasawa Shin, Shobatake Ryogo, Itaya‐Hironaka Asako, Makino Mai, Uchiyama Tomoko, Sakuramoto‐Tsuchida Sumiyo, Takeda Yoshinori, Ota Hiroyo, Yamauchi Akiyo
  • Journal Title: Journal of Cellular and Molecular Medicine Volume: 26 Issue: 24 Pages: 6019-6031
  • DOI: 10.1111/jcmm.17618
  • Peer Reviewed / Open Access
• [Journal Article] Upregulation of Reg IV and Hgf mRNAs by Intermittent Hypoxia via Downregulation of microRNA-499 in Cardiomyocytes (2022)
  • Author(s): Takasawa Shin, Itaya-Hironaka Asako, Makino Mai, Yamauchi Akiyo, Sakuramoto-Tsuchida Sumiyo, Uchiyama Tomoko, Shobatake Ryogo, Takeda Yoshinori, Ota Hiroyo
  • Journal Title: International Journal of Molecular Sciences Volume: 23 Issue: 20 Pages: 12414-12414
  • DOI: 10.3390/ijms232012414
  • Peer Reviewed / Open Access
• [Journal Article] Upregulation of REG IV gene in human intestinal epithelial cells by lipopolysaccharide via downregulation of microRNA‐24 (2022)
  • Author(s): Takasawa Shin, Tsuchida Chikatsugu, Sakuramoto‐Tsuchida Sumiyo, Uchiyama Tomoko, Makino Mai, Yamauchi Akiyo, Itaya‐Hironaka Asako
  • Journal Title: Journal of Cellular and Molecular Medicine Volume: 26 Issue: 17 Pages: 4710-4720
  • DOI: 10.1111/jcmm.17498
  • Peer Reviewed / Open Access
• [Journal Article] Downregulation of the Cd38-Cyclic ADP-Ribose Signaling in Cardiomyocytes by Intermittent Hypoxia via Pten Upregulation (2022)
  • Author(s): Takasawa Shin, Makino Mai, Uchiyama Tomoko, Yamauchi Akiyo, Sakuramoto-Tsuchida Sumiyo, Itaya-Hironaka Asako, Takeda Yoshinori, Asai Keito, Shobatake Ryogo, Ota Hiroyo
  • Journal Title: International Journal of Molecular Sciences Volume: 23 Issue: 15 Pages: 8782-8782
  • DOI: 10.3390/ijms23158782
  • Peer Reviewed / Open Access
• [Journal Article] Identification of fusion transcripts in sarcoma from archival formalin‐fixed paraffin‐embedded tissues: A next‐generation sequencing approach (2022)
  • Author(s): Fujii Tomomi、Takeda Maiko、Uchiyama Tomoko、Nitta Yuji、Maebou Katsuya、Terada Chiyoko、Okada Fumi、Matsuoka Minami、Sugimoto Sumire、Sasaki Shoh、Morita Kohei、Itami Hiroe、Miyake Makito、Takeda Masayuki、Sawabata Noriyoshi、Fujimoto Kiyohide、Ohbayashi Chiho
  • Journal Title: Pathology International Volume: 72 Issue: 9 Pages: 444-456
  • DOI: 10.1111/pin.13265
  • Peer Reviewed
• [Journal Article] Histopathological and epigenetic changes in myocardium associated with cancer therapy-related cardiac dysfunction (2022)
  • Author(s): Chiyoko-Ikeda Terada, Kenji Onoue, Tomomi Fujii, Hiroe Itami, Kohei Morita, Tomoko Uchiyama, Maiko Takeda, Hitoshi Nakagawa, Tomoya Nakano, Youichirou Baba3 , Kisaki Amemiya, Yoshihiko Saito, Kinta Hatakeyama and Chiho Ohbayashi
  • Journal Title: ESC feart failure Volume: 9 Issue: 5 Pages: 3031-3043
  • DOI: 10.1002/ehf2.14034
  • Peer Reviewed
• [Journal Article] Evaluation of DNA/RNA quality from cell block of malignant mesothelioma and lung adenocarcinoma. (2022)
  • Author(s): Tatsumi S, Takeuchi M, Fujii T, Sugimoto A, Sekita-Hatakeyama Y, Nishikawa T, Uchiyama T, Morita K, Itami H, Takeda M, Yamazaki M, Ohbayashi C.
  • Journal Title: Diagn Cytopathol. Volume: 50 Issue: 6 Pages: 273-283
  • DOI: 10.1002/dc.24960
  • Peer Reviewed
• [Journal Article] Carcinoma showing thymus‐like differentiation of the parotid gland: The brief report of cytomorphology and review of the literature (2022)
  • Author(s): Uchiyama Tomoko、Terada Chiyoko、Tachibana Yuma、Nishiura Hirokazu、Takeda Maiko、Fujii Tomomi、Kimura Takahiro、Tsutsumi Masahiro、Ohbayashi Chiho
  • Journal Title: Diagnostic Cytopathology Volume: 51 Issue: 3 Pages: 98-104
  • DOI: 10.1002/dc.25088
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• [Journal Article] Liver metastases of a neuroendocrine tumor arising from a tailgut cyst treated with interventional locoregional therapies: a case report and review of the literature on recurrent cases (2022)
  • Author(s): Fujimoto Kosuke、Koyama Fumikazu、Kuge Hiroyuki、Obara Shinsaku、Iwasa Yosuke、Takei Takeshi、Takagi Tadataka、Sadamitsu Tomomi、Harada Suzuka、Uchiyama Tomoko、Ohbayashi Chiho、Nishiofuku Hideyuki、Tanaka Toshihiro、Sho Masayuki
  • Journal Title: International Cancer Conference Journal Volume: 12 Issue: 2 Pages: 93-99
  • DOI: 10.1007/s13691-022-00587-2
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• [Journal Article] Imaging features of β-catenin-activated hepatocellular adenoma with weak β-catenin activation: A rare case report (2022)
  • Author(s): Minamiguchi Kiyoyuki、Marugami Nagaaki、Uchiyama Tomoko、Kusano Hironori、Yasuda Satoshi、Sho Masayuki、Tanaka Toshihiro
  • Journal Title: Acta Radiologica Open Volume: 11 Issue: 11 Pages: 205846012211422-205846012211422
  • DOI: 10.1177/20584601221142241
  • Peer Reviewed / Open Access
• [Journal Article] Correlation of MTAP Immunohistochemistry With CDKN2A Status Assessed by Fluorescence In Situ Hybridization and Clinicopathological Features in CNS WHO Grade 2 and 3 Meningiomas: A Single Center Cohort Study. (2022)
  • Author(s): Sasaki S, Takeda M, Hirose T, Fujii T, Itami H, Uchiyama T, Morita K, Matsuda R, Yamada S, Nakagawa I, Ohbayashi C.
  • Journal Title: Neuropathol Exp Neurol. Volume: 81 Issue: 2 Pages: 117-126
  • DOI: 10.1093/jnen/nlab127
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• [Journal Article] Evaluation and diagnostic value of next-generation sequencing analysis of residual liquid-based cytology specimens of pancreatic masses. (2022)
  • Author(s): Sekita-Hatakeyama Y, Fujii T, Nishikawa T, Mitoro A, Sawai M, Itami H, Morita K, Uchiyama T, Takeda M, Sho M, Yoshiji H, Hatakeyama K, Ohbayashi C.
  • Journal Title: Cancer Cytopathol. Volume: 130 Issue: 3 Pages: 202-214
  • DOI: 10.1002/cncy.22525
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• [Journal Article] Preference of grade and lymphovascular invasion over invasive size measurement in stage I lung adenocarcinoma (2022)
  • Author(s): Itami Hiroe、Kawaguchi Takeshi、Yoshikawa Daiki、Watanabe Takashi、Terada Chiyoko、Okada Fumi、Uchiyama Tomoko、Takeda Maiko、Ishida Eiwa、Nishimoto Yuko、Okada Hiroshi、Kushibe Keiji、Sawabata Noriyoshi、Ohbayashi Chiho
  • Journal Title: Journal of Clinical Pathology Volume: - Issue: 7 Pages: 486-491
  • DOI: 10.1136/jclinpath-2021-208053
  • Peer Reviewed
• [Journal Article] Effects of Intermittent Hypoxia on Cytokine Expression Involved in Insulin Resistance (2021)
  • Author(s): Uchiyama Tomoko、Ota Hiroyo、Ohbayashi Chiho、Takasawa Shin
  • Journal Title: International Journal of Molecular Sciences Volume: 22 Issue: 23 Pages: 12898-12898
  • DOI: 10.3390/ijms222312898
  • Peer Reviewed / Open Access
• [Journal Article] Diffuse large B-cell lymphoma (DLBCL) with significant intravascular invasion. Close resemblance of its clinicopathological features to intravascular large B-cell lymphoma, but not to DLBCL-not otherwise specified (2021)
  • Author(s): Itami Hiroe、Nakamine Hirokazu、Kubo Masayuki、Ogawa Kohei、Tani Rina、Nakamura Shinji、Takeda Maiko、Nitta Yuji、Uchiyama Tomoko、Fujii Tomomi、Hatakeyama Kinta、Ohbayashi Chiho
  • Journal Title: Journal of Clinical and Experimental Hematopathology Volume: 61 Issue: 3 Pages: 152-161
  • DOI: 10.3960/jslrt.20066
  • NAID: 130008087360
  • ISSN: 1346-4280, 1880-9952
  • Peer Reviewed / Open Access
• [Journal Article] Immunohistochemical Expression Status of p53, CD44v9, and Ki-67 in a Series of Fallopian Tube Lesions of High-grade Serous Carcinoma (2021)
  • Author(s): Sugimoto Sumire、Uchiyama Tomoko、Kawahara Naoki、Ohbayashi Chiho、Kobayashi Hiroshi
  • Journal Title: International Journal of Gynecological Pathology Volume: 40 Issue: 5 Pages: 419-426
  • DOI: 10.1097/pgp.0000000000000738
  • Peer Reviewed
• [Presentation] Intermittent Hypoxia Increased the Expression of Intercellular Adhesion Molecule-1 and Endothelial Cell-Specific Molecule 1 in Vascular Endothelial Cells via the Downregulation of MicroRNA-181a1 (2023)
  • Author(s): Shin Takasawa, Mai Makino, Akiyo Yamauchi, Sumiyo Sakuramoto-Tsuchida, Rina Hirota, Ryusei Fujii, Keito Asai, Yoshinori Takeda, Tomoko Uchiyama, Ryogo Shobatake, Hiroyo Ota
  • Organizer: 83rd ADA Scientific Sessions
  • Int'l Joint Research
• [Presentation] Attenuation of the CD38-cyclic ADP-ribose signaling in cardiomyocytes by intermittent hypoxia via upregulation of Pten (2022)
  • Author(s): Takasawa, S., Makino, M., Yamauchi, A., Sakuramoto-Tsuchida, S., Itaya-Hironaka, A., Uchiyama, T., Takeda, Y., Asai, K., Shobatake, R. and Ota, H
  • Organizer: IDF2022
  • Int'l Joint Research
• [Presentation] Up-regulation of dopamine β-hydroxylase and phenylethanolamine N-methyltransferase by intermittent hypoxia via down-regulation of microRNA-375 in neuronal cells (2022)
  • Author(s): Takasawa, S., Shobatake, R., Takeda, Y., Uchiyama, T., Yamauchi, A., Makino, M., Sakuramoto-Tsuchida, S., Asai, K., Hirota, R., Fujii, R., Ota, H. and Itaya-Hironaka, A
  • Organizer: 58th EASD Annual Meeting
  • Int'l Joint Research
• [Presentation] 自施設における子宮平滑筋肉腫23症例の臨床病理学的検討 (2022)
  • Author(s): 内山智子, 武田麻衣子, 藤井智美, 新田勇治, 岡田文美, 杉本澄美玲, 佐々木翔, 森田剛平, 伊丹弘恵, 大林千穂
  • Organizer: 第111回日本病理学会
• [Presentation] 間歇的低酸素は神経細胞においてカテコールアミン生合成酵素群mRNAを増加させる (2022)
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  • Organizer: 第65回日本糖尿病学会
• [Presentation] FFPEを用いた次世代シークエンス解析による肉腫の融合遺伝子の検出 (2021)
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  • Organizer: 第80回癌学会
• [Presentation] 尿路上皮癌の浸潤増殖能に関わるmiRNA-27a/NACC1の機能解析 (2021)
  • Author(s): 藤井 智美, 内山 智子, 森田 剛平, 伊丹 弘恵, 武田 麻衣子, 島田 啓司, 三宅 牧人, 藤本 清秀, 大林 千穂
  • Organizer: 第110回日本病理学会
• [Book] Advantages of Noncoding RNAs in Molecular Diagnosis (2022)
  • Author(s): Tomomi Fujii, Tomoko Uchiyama and Maiko Takeda
  • Total Pages: 22
  • Publisher: IntechOpen
• [Book] 泌尿器領域 画像診断の勘ドコロNEO (2021)
  • Author(s): 藤井智美、内山智子
  • Total Pages: 312
  • Publisher: 株式会社メジカルビュー社
  • ISBN: 9784758316170