Analysis of the induced cancer stem-like sphere cells-specific molecules to prevent the metastatic properties of hepatocellular carcinom

• Project/Area Number: 16K10574
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Digestive surgery
• Research Institution: Yamaguchi University
• Principal Investigator: TSUNEDOMI Ryouichi 山口大学, 医学部附属病院, 講師 (10420514)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2018: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2017: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2016: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: 癌 / 癌幹細胞

Outline of Final Research Achievements

Cancer stem cells (CSCs) are thought to play important roles in carcinogenesis, recurrence, metastasis, and therapy-resistance. We have successfully induced cancer stem-like sphere cells (CSLCs) from hepatoma cell lines using a unique medium. Our induced CSLCs showed characteristic phenotypes of CSCs such as chemo- and immune-resistances. As a result of integrated analysis with RNA-seq, a RAB3B was identified as an up-regulated gene in both CSLCs in vitro and poor prognostic HCCs in vivo. Cells harboring mono-allelic RAB3B knock-out showed altered sphere formation and significantly lower chemoresistance compared to those of parental cells. The mono-allelic RAB3B knock-out cells also showed decreased PD-1 ligands in sphere induction medium than those in cells cultured with normal medium. Therefore, the up-regulation of RAB3B may plays important roles in CSLCs.

Academic Significance and Societal Importance of the Research Achievements

本研究から，がん幹細胞特性，特に治療抵抗性及び転移能に関与する遺伝子としてRAB3Bを同定した．また，本研究における大量調製可能な誘導がん幹細胞が，免疫逃避機構も備えることが新たに明らかとなり，がん幹細胞を標的としたさらなる免疫療法の開発への展開が可能となった．本研究は，がんの予後不良の主因とも言える術後再発を抑制するための意義ある成果をもたらした．

Research Products

• [Journal Article] Efficacy of perioperative immunonutrition in esophageal cancer patients undergoing esophagectomy (2019)
  • Author(s): Kanekiyo Shinsuke、Takeda Shigeru、Iida Michihisa、Nishiyama Mitsuo、Kitahara Masahiro、Shindo Yoshitaro、Tokumitsu Yukio、Tomochika Shinobu、Tsunedomi Ryoichi、Suzuki Nobuaki、Abe Toshihiro、Yoshino Shigefumi、Hazama Shoichi、Ueno Tomio、Nagano Hiroaki
  • Journal Title: Nutrition Volume: 59 Pages: 96-102
  • DOI: 10.1016/j.nut.2018.08.006
  • Peer Reviewed
• [Journal Article] Overexpression of miR221 and miR222 in the cancer stroma is associated with malignant potential in colorectal cancer (2018)
  • Author(s): Iida Michihisa、Hazama Shoichi、Tsunedomi Ryouichi、Tanaka Hironori、Takenouchi Hiroko、Kanekiyo Shinsuke、Tokumitsu Yukio、Tomochika Shinobu、Tokuhisa Yoshihiro、Sakamoto Kazuhiko、Suzuki Nobuaki、Takeda Shigeru、Ueno Tomio、Yamamoto Shigeru、Yoshino Shigefumi、Fujita Koji、Kuroda Masahiko、Nagano Hiroaki
  • Journal Title: Oncology Reports Volume: 40 Pages: 1621-1631
  • DOI: 10.3892/or.2018.6575
  • Peer Reviewed / Open Access
• [Journal Article] Pharmacogenetics-based area-under-curve model can predict efficacy and adverse events from axitinib in individual patients with advanced renal cell carcinoma (2018)
  • Author(s): Yamamoto Yoshiaki、Tsunedomi Ryouichi、Fujita Yusuke、Otori Toru、Ohba Mitsuyoshi、Kawai Yoshihisa、Hirata Hiroshi、Matsumoto Hiroaki、Haginaka Jun、Suzuki Shigeo、Dahiya Rajvir、Hamamoto Yoshihiko、Matsuyama Kenji、Hazama Shoichi、Nagano Hiroaki、Matsuyama Hideyasu
  • Journal Title: Oncotarget Volume: 9 Issue: 24 Pages: 17160-17170
  • DOI: 10.18632/oncotarget.24715
  • Peer Reviewed / Open Access
• [Journal Article] Hedgehog Signals Mediate Anti-Cancer Drug Resistance in Three-Dimensional Primary Colorectal Cancer Organoid Culture (2018)
  • Author(s): Usui Tatsuya、Sakurai Masashi、Umata Koji、Elbadawy Mohamed、Ohama Takashi、Yamawaki Hideyuki、Hazama Shoichi、Takenouchi Hiroko、Nakajima Masao、Tsunedomi Ryouichi、Suzuki Nobuaki、Nagano Hiroaki、Sato Koichi、Kaneda Masahiro、Sasaki Kazuaki
  • Journal Title: International Journal of Molecular Sciences Volume: 19 Issue: 4 Pages: 1098-1098
  • DOI: 10.3390/ijms19041098
  • Peer Reviewed / Open Access
• [Journal Article] IgG response to MHC class I epitope peptides is a quantitative predictive biomarker in the early course of treatment of colorectal cancer using therapeutic peptides (2018)
  • Author(s): Kanekiyo S, Hazama S, Takenouchi H, Nakajima M, Shindo Y, Matsui H, Tokumitsu Y, Tomochika S, Tsunedomi R, Tokuhisa Y, Iida M, Sakamoto K, Suzuki N, Takeda S, Yamamoto S, Yoshino S, Okuno K, Udaka K, Kawakami Y, Matsueda S, Ito K, Nagano H.
  • Journal Title: Oncology Reports Volume: 39 Pages: 2385-2392
  • DOI: 10.3892/or.2018.6288
  • Peer Reviewed / Open Access
• [Journal Article] Metastatic ability and the epithelial-mesenchymal transition in induced cancer stem-like hepatoma cells (2018)
  • Author(s): Nishiyama M、Tsunedomi R、Yoshimura K、Hashimoto Ni、Matsukuma S、Ogihara H、Kanekiyo S、Iida M、Sakamoto K、Suzuki N、Takeda S、Yamamoto S、Yoshino S、Ueno T、Hamamoto Y、Hazama S、Nagano H
  • Journal Title: Cancer Science Volume: 109 Issue: 4 Pages: 1101-1109
  • DOI: 10.1111/cas.13527
  • Peer Reviewed / Open Access
• [Journal Article] Rapid and sensitive detection of UGT1A1 polymorphisms associated with irinotecan toxicity by a novel DNA microarray (2017)
  • Author(s): Tsunedomi Ryouichi、Hazama Shoichi、Okayama Naoko、Oka Masaaki、Nagano Hiroaki
  • Journal Title: Cancer Science Volume: 108 Issue: 7 Pages: 1504-1509
  • DOI: 10.1111/cas.13272
  • NAID: 130008102268
  • Peer Reviewed / Open Access
• [Journal Article] miR-196b, miR-378a and miR-486 are predictive biomarkers for the efficacy of vaccine treatment in colorectal cancer (2017)
  • Author(s): Shindo Yoshitaro、Hazama Shoichi、Nakamura Yusuke、Inoue Yuka、Kanekiyo Shinsuke、Suzuki Nobuaki、Takenouchi Hiroko、Tsunedomi Ryouichi、Nakajima Masao、Ueno Tomio、Takeda Shigeru、Yoshino Shigefumi、Okuno Kiyotaka、Fujita Yusuke、Hamamoto Yoshihiko、Kawakami Yutaka、Oka Masaaki、Nagano Hiroaki
  • Journal Title: Oncology Letters Volume: 14 Issue: 2 Pages: 1355-1362
  • DOI: 10.3892/ol.2017.6303
  • Peer Reviewed / Open Access
• [Journal Article] miR-125b-1 and miR-378a are predictive biomarkers for the efficacy of vaccine treatment against colorectal cancer (2017)
  • Author(s): Tanaka Hironori、Hazama Shoichi、Iida Michihisa、Tsunedomi Ryouichi、Takenouchi Hiroko、Nakajima Masao、Tokumitsu Yukio、Kanekiyo Shinsuke、Shindo Yoshitaro、Tomochika Shinobu、Tokuhisa Yoshihiro、Sakamoto Kazuhiko、Suzuki Nobuaki、Takeda Shigeru、Yamamoto Shigeru、Yoshino Shigefumi、Ueno Tomio、et. al.
  • Journal Title: Cancer Science Volume: 108 Issue: 11 Pages: 2229-2238
  • DOI: 10.1111/cas.13390
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] High secreted protein acidic and rich in cysteine expression in peritumoral fibroblasts predicts better prognosis in patients with resectable gastric cancer (2017)
  • Author(s): Nakajima Masao、Yoshino Shigefumi、Kanekiyo Shinsuke、Maeda Noriko、Sakamoto Kazuhiko、Tsunedomi Ryoichi、Suzuki Nobuaki、Takeda Shigeru、Yamamoto Shigeru、Hazama Shoichi、Hoshii Yoshinobu、Oga Atsunori、Itoh Hiroshi、Ueno Tomio、Nagano Hiroaki
  • Journal Title: Oncology Letters Volume: 15 Pages: 803-812
  • DOI: 10.3892/ol.2017.7418
  • Peer Reviewed / Open Access
• [Journal Article] MicroRNA-6826 and -6875 in plasma are valuable non invasive biomarkers that predict the efficacy of vaccine treatment against metastatic colorectal cancer. (2017)
  • Author(s): Kijima T, Hazama S, Tsunedomi R, Tanaka H, Takenouchi H, Kanekiyo S, Inoue Y, Nakashima M, Iida M, Sakamoto K, Suzuki N, Takeda S, Ueno T, Yamamoto S, Yoshino S, Okuno K, Nagano H.
  • Journal Title: Oncology Reports Volume: 37 Issue: 1 Pages: 23-30
  • DOI: 10.3892/or.2016.5267
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Cetuximab strongly enhances immune cell infiltration into liver metastatic sites in colorectal cancer. (2017)
  • Author(s): Inoue Y, Hazama S, Suzuki N, Tokumitsu Y, Kanekiyo S, Tomochika S,Tsunedomi R, Tokuhisa Y, Iida M, Sakamoto K, Takeda S, Ueno T, Yoshino S and Nagano H．
  • Journal Title: Cancer Sci. Volume: 108(3) Issue: 3 Pages: 455-460
  • DOI: 10.1111/cas.13162
  • Peer Reviewed / Open Access
• [Journal Article] Calreticulin is highly expressed in pancreatic cancer stem-like cells. (2016)
  • Author(s): Satoshi Matsukuma, Kiyoshi Yoshimura, Tomio Ueno, Atsunori Oga, Moeko Inoue, Yusaku Watanabe, Atsuo Kuramasu, Masanori Fuse, Ryouichi Tsunedomi, Satoshi Nagaoka, Hidetoshi Eguchi, Hiroto Matsui, Yoshitaro Shindo, Noriko Maeda, Yoshihiro Tokuhisa, Reo Kawano, Tomoko Furuya-Kondo, Hiroshi Itoh, Shigefumi Yoshino, et. al.
  • Journal Title: Cancer Science Volume: 107 Issue: 11 Pages: 1599-1609
  • DOI: 10.1111/cas.13061
  • Peer Reviewed / Open Access
• [Journal Article] Prediction of the efficacy of immunotherapy by measuring the integrity of cell-free DNA in plasma in colorectal cancer. (2016)
  • Author(s): Masahiro Kitahara, Shoichi Hazama, Ryouichi Tsunedomi, Hiroko Takenouchi, Shinsuke Kanekiyo, Yuka Inoue, Masao Nakajima, Shinobu Tomochika, Yoshihiro Tokuhisa, Michihisa Iida, Kazuhiko Sakamoto, Nobuaki Suzuki, Shigeru Takeda, Tomio Ueno, Shigeru Yamamoto, Shigefumi Yoshino, Hiroaki Nagano.
  • Journal Title: Cancer Science Volume: 107 Issue: 12 Pages: 1825-1829
  • DOI: 10.1111/cas.13085
  • Peer Reviewed / Open Access
• [Journal Article] Establishment of a novel model for anti-cancer drug resistance in three-dimensional primary culture of tumor microenvironment. (2016)
  • Author(s): Usui T, Sakurai M, Enjoji S, Kawasaki H, Umata K, Ohama T, Fujiwara N, Yabe R, Tsuji S, Yamawaki H, Hazama S, Takenouchi H, Nakajima M, Tsunedomi R, Suzuki N, Nagano H and Sato K.
  • Journal Title: Stem Cells International. Volume: - Issue: 1 Pages: 7053872-7053872
  • DOI: 10.1155/2016/7053872
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] がん幹細胞様細胞と臨床サンプルとの統合解析により同定されたがん幹細胞関連遺伝子 (2018)
  • Author(s): 恒富亮一
  • Organizer: 第26回日本消化器関連学会週間 (JDDW2018)
• [Presentation] Identification of responsible genes for evasion of immune surveillance in induced cancer stem-like cells (2018)
  • Author(s): 恒富亮一
  • Organizer: 第56回日本癌治療学会学術集会
• [Presentation] RAB3B gene was identified as a gene involved in chemoresistance of induced cancer stem-like sphere cells (2018)
  • Author(s): 恒富亮一
  • Organizer: 第77回日本癌学会学術総会
• [Presentation] RAB3B was identified as a gene involved in characteristics of induced cancer stem-like sphere cells (2018)
  • Author(s): 恒富亮一
  • Organizer: 23rd World Congress on Advances in Oncology and 22nd International Symposium on Molecular Medicine
  • Int'l Joint Research
• [Presentation] 誘導癌幹細胞における免疫逃避機構の検討 (2018)
  • Author(s): 恒富亮一
  • Organizer: 第73回日本消化器外科学会総会
• [Presentation] がん幹細胞様細胞におけるCD44 isoforms発現 (2018)
  • Author(s): 恒富亮一
  • Organizer: 第6回がんと代謝研究会
• [Presentation] 肝転移能亢進を示す肝細胞癌幹細胞様細胞の特性 (2017)
  • Author(s): 恒富 亮一、吉村 清、西山 光郎、松隈 聰、徳光 幸生、兼清 信介、友近忍、徳久 善弘、飯田 通久、坂本 和彦、鈴木 伸明、武田 茂、山本 滋、吉野 茂文、硲 彰一、上野 富雄，永野 浩昭
  • Organizer: 第26回日本がん転移学会学術集会・総会
• [Presentation] Identification of genes involved in chemoresistance of induced cancer stem-like sphere cells (2017)
  • Author(s): Tsunedomi Ryouichi, Yoshimura Kiyoshi, Matsukuma Satoshi, Kanekiyo Shinsuke, Michihisa Iida, Suzuki Nobuaki, Takeda Shigeru, Yoshino Shigefumi, Hazama Shoichi, Ueno Tomio, Nagano Hiroaki
  • Organizer: 第76回日本癌学会学術総会
• [Presentation] 膵癌幹細胞様細胞に高発現した分子の同定と解析 (2017)
  • Author(s): 松隈 聰、吉村 清、坂本 和彦、小賀 厚徳、恒富 亮一、布施 雅規、鈴木 伸明、武田 茂、吉野 茂文、硲 彰一、江口 英利、上野 富雄、永野 浩昭
  • Organizer: 第76回日本癌学会学術総会
• [Presentation] 膵癌幹細胞の新たな表面マーカーcalreticulinの同定 (2017)
  • Author(s): 松隈 聰, 吉村 清, 江口 英利, 恒富 亮一, 松井 洋人, 兼清 信介, 徳光 幸生, 友近 忍, 徳久 善弘, 飯田 通久, 坂本 和彦, 鈴木 伸明, 武田 茂, 吉野 茂文, 硲 彰一, 上野 富雄, 永野 浩昭
  • Organizer: 第25回日本消化器関連学会週間
• [Presentation] 癌幹細胞様細胞における抗癌剤耐性に影響を及ぼす遺伝子の同定 (2017)
  • Author(s): 恒富亮一、吉村 清、松隈 聰、兼清信介、徳光幸生、友近 忍、徳久善弘、飯田通久、坂本和彦、鈴木伸明、武田 茂、山本 滋、吉野茂文、硲 彰一、上野富雄、永野浩昭
  • Organizer: 第55回日本癌治療学会学術集会
• [Presentation] RESPONSIBLE GENES FOR CHARACTERISTICS OF THE INDUCED CANCER STEM-LIKE SPHERE CELLS OF HEPATOCELLULAR CARCINOMA (2017)
  • Author(s): Ryouichi Tsunedomi, Kiyoshi Yoshimura, Mitsuo Nishiyama, Satoshi Matsukuma, Shinsuke Kanekiyo, Yukio Tokumitsu, Shinobu Tomochika, Yoshihiro Tokuhisa, Michihisa Iida, Kazuhiko Sakamoto, Nobuaki Suzuki, Shigeru Takeda, Shigeru Yamamoto, Shigehumi Yoshino, Shoichi Hazama, Tomio Ueno, Hiroaki Nagano
  • Organizer: 25th United European Gastroenterology Week
  • Int'l Joint Research
• [Presentation] 癌幹細胞様特性を示すSphere 細胞における抗癌剤耐性に関与する遺伝子の同定 (2017)
  • Author(s): 恒富 亮一、西山 光郎、藤原 信行、松隈 聰、兼清 信介、徳久 善弘、徳光 幸生、飯田 通久、坂本 和彦、鈴木 伸明、武田 茂、山本 滋、吉野 茂文、硲 彰一、上野 富雄、永野 浩昭
  • Organizer: 第28回日本消化器癌発生学会総会
• [Presentation] 肝転移能亢進を示す癌幹細胞様細胞におけるCD44 isoformsの発現 (2016)
  • Author(s): 恒富 亮一、松隈 聰、坂本 和彦、徳久 善弘、徳光 幸生、飯田 通久、鈴木 伸明、武田 茂、上野 富雄、吉野 茂文、硲 彰一、永野 浩昭
  • Organizer: 第27回日本消化器癌発生学会総会
  • Place of Presentation: 城山観光ホテル (鹿児島)
• [Presentation] 膵癌幹細胞様細胞に高発現する分子の同定と同分子の臨床的意義に関する検討 (2016)
  • Author(s): 松隈 聰、吉村 清、小賀 厚徳、井上 萌子、倉増 敦朗、布施 雅規、渡邊裕策、恒富 亮一、長岡 彗、江口 英利、松井 洋人、徳光 幸生、徳久 善弘、坂本 和彦、近藤 智子、上野 富雄、吉野 茂文、硲 彰一、伊藤 浩史、永野 浩昭
  • Organizer: 第27回日本消化器癌発生学会総会
  • Place of Presentation: 城山観光ホテル (鹿児島)
• [Presentation] 癌間質におけるmiR221およびmiR222過剰発現は大腸癌の悪性度と相関する (2016)
  • Author(s): 飯田 通久、恒富 亮一、田中 宏典、兼清 信介、徳久 善弘、鈴木 伸明、武田 茂、上野 富雄、硲 彰一、吉野 茂文、永野 浩昭
  • Organizer: 第27回日本消化器癌発生学会総会
  • Place of Presentation: 城山観光ホテル (鹿児島)
• [Presentation] Metastatic potentials and expressions of CD44 isoforms in hepatocellular carcinoma stem-like sphere cells (2016)
  • Author(s): Ryouichi Tsunedomi, Satoshi Matsukuma, Shinsuke Kanekiyo, Kiyoshi Yoshimura, Nobuaki Suzuki, Sigeru Takeda, Tomio Ueno, Shigefumi Yoshino, Shoichi Hazama, Hiroaki Nagano
  • Organizer: 第75回日本癌学会学術総会
  • Place of Presentation: パシフィコ横浜 (神奈川)
• [Presentation] Calreticulin highly expressed in pancreatic cancer stem-like cells is a prognostic factor for pancreatic cancer (2016)
  • Author(s): Satoshi Matsukuma, Kiyoshi Yoshimura, Tomio Ueno, Atsunori Oga, Moeko Inoue, Atsuo Kuramasu, Masanori Fuse, Ryoichi Tsunedomi, Hidetoshi Eguchi, Hiroto Matsui, Shigefumi Yoshino, Shoichi Hazama, Hiroaki Nagano
  • Organizer: 第75回日本癌学会学術総会
  • Place of Presentation: パシフィコ横浜 (神奈川)
• [Presentation] Overexpression of miR221/222 in cancer stroma are associated with malignant potentials in colorectal cancer (2016)
  • Author(s): Michihisa Iida, Shoichi Hazama, Ryoichi Tsunedomi, Shigeru Takeda, Tomio Ueno, Shigeru Yamamoto, Shigefumi Yoshino, Hiroaki Nagano
  • Organizer: 第75回日本癌学会学術総会
  • Place of Presentation: パシフィコ横浜 (神奈川)
• [Presentation] 肝細胞癌 (HCC) に対する術後補助療法としてのHSP70-mRNA導入樹状細胞療法I/II相試験 (2016)
  • Author(s): 松井 洋人、硲 彰一、中島 正夫、松隈聰、井上 由佳、兼清 信介、徳光 幸生、新藤 芳太郎、恒富 亮一、友近 忍、飯田 通久、徳久 善弘、坂本 和彦、鈴木 伸明、武田 茂、山本 滋、上野 富雄、吉野 茂文、永野 浩昭
  • Organizer: 第29回日本バイオセラピィ学会学術集会総会
  • Place of Presentation: 久留米シティプラザ (福岡)
• [Presentation] 大腸癌ワクチン療法におけるmiR125b-1, miR-378aの治療効果予測マーカーとしての有用性 ~Laser capture microdissection法による癌細胞・間質細胞分離の応用~ (2016)
  • Author(s): 田中 宏典、硲 彰一、松井 洋人、徳光 幸生、兼清 信介、友近 忍、飯田 通久、恒富 亮一、徳久 善弘、坂本 和彦、鈴木 伸明、武田 茂、上野 富雄、山本 茂、吉野 茂文、藤田 浩司、黒田 雅彦、奥野 清隆、河上 裕、永野 浩昭
  • Organizer: 第29回日本バイオセラピィ学会学術集会総会
  • Place of Presentation: 久留米シティプラザ (福岡)
• [Patent(Industrial Property Rights)] 癌幹細胞における薬物耐性の低減剤、癌幹細胞における転移能の抑制剤及び癌の転移性再発リスクを予測する方法 (2018)
  • Inventor(s): 恒富亮一、永野浩昭
  • Industrial Property Rights Holder: 国立大学法人山口大学
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2018-153470
  • Filing Date: 2018
• [Patent(Industrial Property Rights)] 癌細胞における免疫チェックポイント因子の発現抑制剤及び癌治療用医薬組成物 (2018)
  • Inventor(s): 恒富亮一、永野浩昭
  • Industrial Property Rights Holder: 国立大学法人山口大学
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2018-173611
  • Filing Date: 2018
• [Patent(Industrial Property Rights)] 癌幹細胞における薬物耐性の低減剤及び癌の転移性再発リスクを予測する方法 (2017)
  • Inventor(s): 恒富 亮一、永野 浩昭
  • Industrial Property Rights Holder: 山口大学
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2017-157349
  • Filing Date: 2017