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 – 2025-03-31
• Project Status: Granted (Fiscal Year 2023)
• 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

子宮平滑筋肉腫について、腫瘍の発生や増殖に関与する候補分子を推定し、分子生物学的機序に貢献するmicroRNAの特定とその発現による機能的役割を解明するため、1.FFPEを用いたATRXタンパク・mRNA・miRNA発現の検討、2.テロメア特異的FISH法によるALTの検討、3.子宮平滑筋肉腫細胞を用いた、ATRX発現解析及び分子生物学的機能解析、4.miRNA発現とATRX mRNA発現に与える影響の検証、について検討を進めてきた。今年度は引き続き、平滑筋肉腫細胞株（SK-UT-1, SK-LMS-1, SKN）を用いたin vitroの実験により上皮間葉移行が生じていることが推定されたmiR-367,675の機能解析を進めた。特に影響の強かったmiR-367に着目し、Target scan for Human(Release8.0:2021)を用いて、選択した推定標的候補分子のHIPK3, MYO1B, RAB23遺伝子の発現動態をそれぞれのsiRNAを導入し検討した。平滑筋肉腫由来の細胞株であるSK-LMS-1, SKNにおいては、この3つの遺伝子がmiR-367の制御を受けるとともにRAB23を上流としてMYO1B, HIPK3の発現を調節していることからこれらの分子の増殖能、浸潤能に着目し機能解析を行ったところ、MTS assayではsiRNAを用いたHIPK3, MYO1B, RAB23の抑制によって細胞増殖能は低下した。浸潤能についてInvasion assayによって検討したところ、Matrigel assayではmiR-367の過剰発現およびHIPK3, MYO1B, RAB23の抑制によって種々の程度に浸潤能は抑制された。この結果からこれらの分子が細胞増殖能や浸潤能に直接的に関与していることが明らかとなった。

Current Status of Research Progress

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

Reason

NGSを用いた解析結果を現在論文にまとめているところである。細胞実験についても今後論文化を予定している。

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う。HIPK3，MYO1BおよびRAB23といずれも子宮平滑筋肉腫において増殖能、浸潤能いずれも直接関与していることから、これらの分子の相互作用や特に重要な機能についてさらに解析を進める。
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] Hepatobiliary-phase gadolinium ethoxybenzyl-diethylenetriaminepentaacetic acid MRI for pretreatment prediction of efficacy-to-standard-therapies based on Barcelona Clinic Liver Cancer algorithm: an up-to-date review (2023)
  • Author(s): Minamiguchi Kiyoyuki、Irizato Mariko、Uchiyama Tomoko、Taiji Ryosuke、Nishiofuku Hideyuki、Marugami Nagaaki、Tanaka Toshihiro
  • Journal Title: European Radiology Volume: 33 Issue: 12 Pages: 8764-8775
  • DOI: 10.1007/s00330-023-09950-0
  • Peer Reviewed
• [Journal Article] Intermittent hypoxia increased the expression of <scp>ESM1</scp> and <scp>ICAM‐1</scp> in vascular endothelial cells via the downregulation of <scp>microRNA</scp>‐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
  • Journal Title: Journal of Cellular and Molecular Medicine Volume: 28 Issue: 1
  • DOI: 10.1111/jcmm.18039
  • Peer Reviewed / Open Access
• [Journal Article] Overexpression of MicroRNA-138 Affects the Proliferation and Invasion of Urothelial Carcinoma Cells by Suppressing SOX9 Expression (2023)
  • Author(s): Nitta Yuji、Fujii Tomomi、Uchiyama Tomoko、Sugimoto Aya、Nishikawa Takeshi、Takeda Maiko、Miyake Makito、Shimada Keiji、Fujimoto Kiyohide
  • Journal Title: Biomedicines Volume: 11 Issue: 11 Pages: 3064-3064
  • DOI: 10.3390/biomedicines11113064
  • Peer Reviewed / Open Access
• [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
  • Peer Reviewed
• [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
  • Peer Reviewed / Open Access
• [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
  • Peer Reviewed / Open Access
• [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
  • Peer Reviewed
• [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
  • Peer Reviewed
• [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
  • Peer Reviewed
• [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] 間歇的低酸素はヒト単球細胞においてmicroRNA-146a低下を介してIL-1β, IL-6, IL-8を増加させる (2024)
  • Author(s): 高沢 伸, 牧野 舞, 正畠 良悟, 山内 晶世, 土田 澄代, 内山智子, 竹田善紀, 水田啓太, 朝井啓斗, 太田 浩世
  • Organizer: 第67回日本糖尿病学会年次学術集会
• [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): Takasawa, S., Makino, M., Yamauchi, A., Sakuramoto-Tsuchida, S., Hirota, R., Fujii, R., Asai, K., Takeda, Y., Uchiyama, T., Shobatake, R. and Ota, H.
  • Organizer: 83rd ADA Scientific Sessions, San Diego, CA,
  • Int'l Joint Research
• [Presentation] Up-regulation of ICAM-1 and ESM 1 by intermittent hypoxia via down-regulation of microRNA-181a1 in vascular endothelial cells (2023)
  • Author(s): S. Takasawa, M. Makino, A. Yamauchi, S. Sakuramoto-Tsuchida, R. Hirota, R. Fujii, K. Asai, Y. Takeda, T. Uchiyama, R. Shobatake, H. Ota
  • Organizer: the 59th EASD Annual Meeting
  • Int'l Joint Research
• [Presentation] Gene expression of cardiomyocytes by intermittent hypoxia: the Cd38-cyclic ADP-ribose (cADPR) signaling for functioning and the Reg-Reg receptor system for regeneration. (2023)
  • Author(s): Takasawa, S., Makino, M., Uchiyama, T., Yamauchi, A., Sakuramoto-Tsuchida, S., Itaya-Hironaka, A., Takeda, Y., Asai, K., Shobatake, R. and Ota, H.
  • Organizer: 4th Edition of World Congress on ENDOCRINOLOGY, DIABETES AND METABOLISM
  • Int'l Joint Research
• [Presentation] 間歇的低酸素は血管内皮細胞でmicroRNA-181a1の発現低下を介してICAM-1, ESM1の発現増加を引き起こす (2023)
  • Author(s): 高沢 伸1,2、牧野 舞1、山内晶世1、土田澄代1、廣田梨奈1、藤井龍星1、朝井啓斗1、竹田善紀1,3、内山智子1,2、正畠良悟1,4、太田浩世1,5
  • Organizer: 第96回日本生化学会大会
• [Presentation] 間歇的低酸素は血管内皮細胞のICAM-1, ESM1 mRNAを増加させる (2023)
  • Author(s): 高沢 伸、牧野 舞、山内晶世、土田澄代、廣田梨奈、藤井龍星、朝井啓斗、竹田善紀、内山智子、正畠良悟、太田浩世
  • Organizer: 第66回日本糖尿病学会年次学術集会
• [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)
  • Author(s): 高沢 伸, 正畠 良悟, 竹田 善紀,内山智子, 山内 晶世, 牧野 舞,土田 澄代, 朝井啓斗, 太田 浩世, 広中 安佐子
  • 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)
  • Author(s): 藤井智美、武田麻衣子、内山智子、森田剛平、三宅牧人、大林千穂
  • 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