骨肉腫幹細胞モデルと微小環境のクロストークの解明とそれらを標的にした治療戦略

• Project/Area Number: 21K09317
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 56020:Orthopedics-related
• Research Institution: Kanazawa University
• Principal Investigator: 武内 章彦 金沢大学, 医学系, 協力研究員 (70512218)
• Co-Investigator(Kenkyū-buntansha): 山本 靖彦 金沢大学, 医学系, 教授 (20313637)
• Project Period (FY): 2021-04-01 – 2024-03-31
• Project Status: Completed (Fiscal Year 2023)
• Budget Amount: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000); Fiscal Year 2023: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2022: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2021: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
• Keywords: 骨肉腫 / がん幹細胞 / エクソソーム / 微小環境

Outline of Research at the Start

骨肉腫幹細胞モデルが、微小環境を形成する細胞にどのように作用しているかの解析を進めるとともに、mir34a、Wnt5aそれぞれの作用も解析する。また、免疫系保持マウスにマウス骨肉腫細胞を移植し、合成mir34aとWnt5a阻害剤および免疫チェックポイント阻害剤(ICI) やマルチキナーゼ阻害剤(MKI)で治療し、微小環境を含めた効果を解析する。さらに、臨床検体において、微小環境を形成している細胞と臨床経過・予後との関連があるかを明らかにすることで、新たな病態解明および治療ターゲットの創出を行うことが研究目的である

Outline of Annual Research Achievements

これまでにRAGE発現により誘導される骨肉腫細胞の幹細胞化がmiR34aの発現低下とWNT5aの高発現によることを明らかにしてきた。本研究では、この骨肉腫細胞の幹細胞化機構について、腫瘍細胞と腫瘍微小環境の両視点から研究を進める。昨年度までに、マウス骨肉腫細胞の一つであるDunn細胞は、肺高転移株LM8細胞より膜型RAGE（mRAGE）および内在性分泌型RAGE（esRAGE）の発現が高く、スフェロイド形成能についても高いことを明らかにした。LM8細胞にmRAGEを過剰発現させたLM8-RAGE細胞の腫瘍形成能は、対照のLM8-mock細胞よりも高いことが分かった。ヒト骨肉腫組織（手術検体）を用いてmiR34a発現の解析を行ったが、臨床経過、原発巣や転移巣でのmiR34a陽性率には有意な相関は見られなかった。しかし、miR34a発現パターンが症例ごとに腫瘍内と周辺組織で異なっていることが分かった。
以上の結果をふまえ、本年度はmRAGE高発現Dunn（Dunn-RAGE）細胞とその対照のDunn-mock細胞の皮下移植一次腫瘍形成能、スフェロイド形成能、miR34aおよびWnt5a発現の検討を行った。その結果、Dunn-RAGE細胞の一次腫瘍形成能とスフェロイド形成能はDunn-mock細胞よりも有意に高く、Dunn-RAGE細胞においてmiR34aの発現低下とともにWnt5aの発現亢進が観察できた。続いてDunn-RAGE細胞とDunn-mock細胞のエクソソーム中のmiR34a発現について解析を行ったが、予想に反して両細胞において有意な差は観察されなかった。
今後、骨肉腫細胞に加え、腫瘍微小環境形成細胞のエクソソームの網羅的解析、腫瘍細胞と線維芽細胞などとの共培養の解析を行い、骨肉腫細胞の幹細胞化の分子機構を明らかにし、新たな骨肉腫治療戦略の創出へつなげていく予定である。

Research Products

• [Journal Article] Graft Survivals after Reconstruction Using Tumor-Bearing Frozen Bone in the Extremities (2023)
  • Author(s): Miwa Shinji、Yamamoto Norio、Hayashi Katsuhiro、Takeuchi Akihiko、Igarashi Kentaro、Bangcoy Martin Louie S.、Taniguchi Yuta、Morinaga Sei、Asano Yohei、Tsuchiya Hiroyuki
  • Journal Title: Cancers Volume: 15 Issue: 15 Pages: 3926-3926
  • DOI: 10.3390/cancers15153926
  • Peer Reviewed / Open Access
• [Journal Article] What Are the Complications, Function, and Survival of Tumor-devitalized Autografts Used in Patients With Limb-sparing Surgery for Bone and Soft Tissue Tumors? A Japanese Musculoskeletal Oncology Group Multi-institutional Study (2023)
  • Author(s): Takeuchi Akihiko、Tsuchiya Hiroyuki、Setsu Nokitaka、Gokita Tabu、Tome Yasunori、Asano Naofumi、Minami Yusuke、Kawashima Hiroyuki、Fukushima Suguru、Takenaka Satoshi、Outani Hidetatsu、Nakamura Tomoki、Tsukushi Satoshi、Kawamoto Teruya、et al
  • Journal Title: Clinical Orthopaedics & Related Research Volume: 481 Issue: 11 Pages: 2110-2124
  • DOI: 10.1097/corr.0000000000002720
  • Peer Reviewed
• [Journal Article] Needle tract seeding of a sclerosing epithelioid fibrosarcoma in a biopsy tract: a case report (2023)
  • Author(s): Kawai Masafumi、Miwa Shinji、Yamamoto Norio、Hayashi Katsuhiro、Takeuchi Akihiko、Igarashi Kentaro、Taniguchi Yuta、Araki Yoshihiro、Yonezawa Hirotaka、Nojima Takayuki、Tsuchiya Hiroyuki
  • Journal Title: BMC Musculoskeletal Disorders Volume: 24 Issue: 1 Pages: 454-454
  • DOI: 10.1186/s12891-023-06553-0
• [Journal Article] Clinical Outcome of Low-Grade Myofibroblastic Sarcoma in Japan: A Multicenter Study from the Japanese Musculoskeletal Oncology Group (2023)
  • Author(s): Kito Munehisa、Ae Keisuke、Okamoto Masanori、Endo Makoto、Ikuta Kunihiro、Takeuchi Akihiko、Yasuda Naohiro、Yasuda Taketoshi、Imura Yoshinori、Morii Takeshi、Kikuta Kazutaka、Kawamoto Teruya、Nezu Yutaka、Baba Ichiro、Ohshika Shusa、Uehara Takeshi、Ueda Takafumi、Takahashi Jun、Kawano Hirotaka
  • Journal Title: Cancers Volume: 15 Issue: 8 Pages: 2314-2314
  • DOI: 10.3390/cancers15082314
  • Peer Reviewed / Open Access
• [Journal Article] A validation study for the utility of serum microRNA as a diagnostic and prognostic marker in patients with osteosarcoma (2023)
  • Author(s): Yoshihiro Araki, Naofumi Asano, Norio Yamamoto, Katsuhiro Hayashi, Akihiko Takeuchi, Shinji Miwa, Kentaro Igarashi, Takashi Higuchi, Kensaku Abe, Yuta Taniguchi, Hirotaka Yonezawa, Sei Morinaga, Yohei Asano, Takeshi Yoshida, Rikinari Hanayama, Juntaro Matsuzaki, Takahiro Ochiya, Akira Kawai, Hiroyuki Tsuchiya
  • Journal Title: Oncology Letters Volume: 25(6); 222 Issue: 6 Pages: 222-222
  • DOI: 10.3892/ol.2023.13808
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Clinical Benefits of LigaSure? for Surgical Amputation Without Tourniquet Use in Patients With Musculoskeletal Tumor (2023)
  • Author(s): BANGCOY MARTIN LOUIE S.、MIWA SHINJI、YAMAMOTO NORIO、HAYASHI KATSUHIRO、TAKEUCHI AKIHIKO、IGARASHI KENTARO、TANIGUCHI YUTA、MORINAGA SEI、ASANO YOHEI、TSUCHIYA HIROYUKI
  • Journal Title: Anticancer Research Volume: 43 Issue: 5 Pages: 2127-2133
  • DOI: 10.21873/anticanres.16374
• [Journal Article] Antibacterial effect and biological reaction of calcium phosphate cement impregnated with iodine for use in bone defects (2023)
  • Author(s): Morinaga Sei、Yamamoto Norio、Tokoro Masaharu、Hayashi Katsuhiro、Takeuchi Akihiko、Miwa Shinji、Igarashi Kentaro、Taniguchi Yuta、Asano Yohei、Nojima Takayuki、Tsuchiya Hiroyuki
  • Journal Title: Journal of Biomaterials Applications Volume: 37 Issue: 10 Pages: 1716-1723
  • DOI: 10.1177/08853282231164827
  • Peer Reviewed
• [Journal Article] A nonsteroidal anti-inflammatory drug, zaltoprofen, inhibits the growth of extraskeletal chondrosarcoma cells by inducing PPARγ, p21, p27, and p53 (2022)
  • Author(s): Higuchi T, Takeuchi A, Munesue S, Yamamoto N, Hayashi K, Harashima A, Yamamoto Y, Tsuchiya H
  • Journal Title: Cell Cycle Volume: 22 Issue: 8 Pages: 939-950
  • DOI: 10.1080/15384101.2023.2166195
  • Peer Reviewed / Open Access
• [Journal Article] Primary Osteosarcoma in Patients Over 75 Years of Age: A Case Series and Literature Review (2022)
  • Author(s): ARAKI YOSHIHIRO、YAMAMOTO NORIO、HAYASHI KATSUHIRO、TAKEUCHI AKIHIKO、MIWA SHINJI、IGARASHI KENTARO、HIGUCHI TAKASHI、ABE KENSAKU、TANIGUCHI YUTA、YONEZAWA HIROTAKA、MORINAGA SEI、ASANO YOHEI、NOJIMA TAKAYUKI、TSUCHIYA HIROYUKI
  • Journal Title: In Vivo Volume: 36 Issue: 6 Pages: 2899-2909
  • DOI: 10.21873/invivo.13031
  • Peer Reviewed
• [Journal Article] Pretreatment Neutrophil Count and Platelet-lymphocyte Ratio as Predictors of Metastasis in Patients With Osteosarcoma (2022)
  • Author(s): ARAKI YOSHIHIRO、YAMAMOTO NORIO、HAYASHI KATSUHIRO、TAKEUCHI AKIHIKO、MIWA SHINJI、IGARASHI KENTARO、HIGUCHI TAKASHI、ABE KENSAKU、TANIGUCHI YUTA、YONEZAWA HIROTAKA、MORINAGA SEI、ASANO YOHEI、TSUCHIYA HIROYUKI
  • Journal Title: Anticancer Research Volume: 42 Issue: 2 Pages: 1081-1089
  • DOI: 10.21873/anticanres.15570
  • Peer Reviewed
• [Journal Article] Clinical Outcomes of Soft Tissue Sarcoma around the Elbow Joint: A Retrospective Single Institution Study (2022)
  • Author(s): Azurin Edelyn S.、Yamamoto Norio、Hayashi Katsuhiro、Takeuchi Akihiko、Tada Kaoru、Miwa Shinji、Igarashi Kentaro、Higuchi Takashi、Yonezawa Hirotaka、Morinaga Sei、Asano Yohei、Saito Shiro、Tsuchiya Hiroyuki
  • Journal Title: Sarcoma Volume: 2022 Pages: 1-8
  • DOI: 10.1155/2022/1087726
  • Peer Reviewed / Open Access
• [Journal Article] Monostotic Fibrous Dysplasia in the Femur Strongly Expressing RANKL With Concomitant Osteoporotic Vertebral Compression Fracture: A Case Report (2022)
  • Author(s): AZURIN EDELYN S.、YAMAMOTO NORIO、HAYASHI KATSUHIRO、TAKEUCHI AKIHIKO、MIWA SHINJI、IGARASHI KENTARO、HIGUCHI TAKASHI、YONEZAWA HIROTAKA、MORINAGA SEI、ASANO YOHEI、SAITO SHIRO、TSUCHIYA HIROYUKI
  • Journal Title: Cancer Diagnosis & Prognosis Volume: 2 Issue: 1 Pages: 93-100
  • DOI: 10.21873/cdp.10082
  • Peer Reviewed / Open Access
• [Journal Article] Clinical Outcomes of Limb-sparing Tumor Surgery With Vascular Reconstruction for Bone and Soft-tissue Tumors (2022)
  • Author(s): ASANO YOHEI、YAMAMOTO NORIO、HAYASHI KATSUHIRO、TAKEUCHI AKIHIKO、MIWA SHINJI、IGARASHI KENTARO、TANIGUCHI YUTA、MORINAGA SEI、TADA KAORU、NOJIMA TAKAYUKI、TSUCHIYA HIROYUKI
  • Journal Title: Anticancer Research Volume: 42 Issue: 9 Pages: 4619-4626
  • DOI: 10.21873/anticanres.15966
  • Peer Reviewed
• [Journal Article] The Therapeutic Effect and Clinical Outcome of Immune Checkpoint Inhibitors on Bone Metastasis in Advanced Non-Small-Cell Lung Cancer (2022)
  • Author(s): Asano Yohei、Yamamoto Norio、Demura Satoru、Hayashi Katsuhiro、Takeuchi Akihiko、Kato Satoshi、Miwa Shinji、Igarashi Kentaro、Higuchi Takashi、Yonezawa Hirotaka、Araki Yoshihiro、Morinaga Sei、Saito Shiro、Sone Takashi、Kasahara Kazuo、Tsuchiya Hiroyuki
  • Journal Title: Frontiers in Oncology Volume: 12 Pages: 871675-871675
  • DOI: 10.3389/fonc.2022.871675
  • Peer Reviewed / Open Access
• [Journal Article] Inhibitory Effects of Saururus chinensis Extract on Receptor for Advanced Glycation End-Products-Dependent Inflammation and Diabetes-Induced Dysregulation of Vasodilation (2022)
  • Author(s): Hayashi Kenjiro、Sato Koichi、Ochi Seishi、Kawano Shuhei、Munesue Seiichi、Harashima Ai、Oshima Yu、Kimura Kumi、Kyoi Takashi、Yamamoto Yasuhiko
  • Journal Title: International Journal of Molecular Sciences Volume: 23 Issue: 10 Pages: 5757-5757
  • DOI: 10.3390/ijms23105757
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Soluble receptor for advanced glycation end products protects from ischemia- and reperfusion-induced acute kidney injury (2022)
  • Author(s): Miyagawa Taro、Iwata Yasunori、Oshima Megumi、Ogura Hisayuki、Sato Koichi、Nakagawa Shiori、Yamamura Yuta、Kamikawa Yasutaka、Miyake Taito、Kitajima Shinji、Toyama Tadashi、Hara Akinori、Sakai Norihiko、Shimizu Miho、Furuichi Kengo、Munesue Seiichi、Yamamoto Yasuhiko、Kaneko Shuichi、Wada Takashi
  • Journal Title: Biology Open Volume: 11 Issue: 1 Pages: 1-8
  • DOI: 10.1242/bio.058852
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] A scoring system combining clinical, radiological, and histopathological examinations for differential diagnosis between lipoma and atypical lipomatous tumor/well-differentiated liposarcoma (2022)
  • Author(s): Asano Yohei、Miwa Shinji、Yamamoto Norio、Hayashi Katsuhiro、Takeuchi Akihiko、Igarashi Kentaro、Yonezawa Hirotaka、Araki Yoshihiro、Morinaga Sei、Nojima Takayuki、Ikeda Hiroko、Tsuchiya Hiroyuki
  • Journal Title: Scientific Reports Volume: 12 Issue: 1 Pages: 237-237
  • DOI: 10.1038/s41598-021-04004-1
  • Peer Reviewed / Open Access
• [Journal Article] Therapeutic Targets and Emerging Treatments in Advanced Chondrosarcoma (2022)
  • Author(s): Miwa Shinji、Yamamoto Norio、Hayashi Katsuhiro、Takeuchi Akihiko、Igarashi Kentaro、Tsuchiya Hiroyuki
  • Journal Title: International Journal of Molecular Sciences Volume: 23 Issue: 3 Pages: 1096-1096
  • DOI: 10.3390/ijms23031096
  • Peer Reviewed / Open Access
• [Journal Article] Primary total knee arthroplasty assisted by computed tomography-free navigation for secondary knee osteoarthritis following massive calcium phosphate cement packing for distal femoral giant-cell bone tumor treatment: a case report (2022)
  • Author(s): Takeuchi A, Yamamoto N, Ohmori T, Hayashi K, Miwa S, Igarashi K, Higuchi T, Abe K, Yonezawa H, Morinaga S, Araki Y, Asano Y, Saito S, Tsuchiya H.
  • Journal Title: BMC Musculoskeletal Disorders Volume: 22 Issue: 1 Pages: 170-170
  • DOI: 10.1186/s12891-022-05131-0
  • Peer Reviewed / Open Access
• [Journal Article] Impact of Tumor Growth Speed of Primary Lesions on the Clinical Outcomes of Appendicular Skeletal Metastases (2021)
  • Author(s): ASANO YOHEI、YAMAMOTO NORIO、HAYASHI KATSUHIRO、TAKEUCHI AKIHIKO、MIWA SHINJI、IGARASHI KENTARO、YONEZAWA HIROTAKA、ARAKI YOSHIHIRO、MORINAGA SEI、SAITO SHIRO、NOJIMA TAKAYUKI、TSUCHIYA HIROYUKI
  • Journal Title: Anticancer Research Volume: 42 Issue: 1 Pages: 229-236
  • DOI: 10.21873/anticanres.15477
  • Peer Reviewed
• [Journal Article] Clinical characteristics of patients with undergoing unplanned excisions of malignant soft tissue tumors (2021)
  • Author(s): Morinaga Sei、Miwa Shinji、Yamamoto Norio、Hayashi Katsuhiro、Takeuchi Akihiko、Igarashi Kentaro、Tada Kaoru、Langit Mickhael B、Yonezawa Hirotaka、Araki Yoshihiro、Asano Yohei、Tsuchiya Hiroyuki
  • Journal Title: Journal of Orthopaedic Surgery Volume: 29 Issue: 3 Pages: 1-12
  • DOI: 10.1177/23094990211057597
  • Peer Reviewed / Open Access
• [Journal Article] A Radiological Scoring System for Differentiation between Enchondroma and Chondrosarcoma (2021)
  • Author(s): Miwa Shinji、Yamamoto Norio、Hayashi Katsuhiro、Takeuchi Akihiko、Igarashi Kentaro、Tada Kaoru、Yonezawa Hirotaka、Morinaga Sei、Araki Yoshihiro、Asano Yohei、Saito Shiro、Okuda Miho、Taki Junichi、Ikeda Hiroko、Nojima Takayuki、Tsuchiya Hiroyuki
  • Journal Title: Cancers Volume: 13 Issue: 14 Pages: 3558-3558
  • DOI: 10.3390/cancers13143558
  • Peer Reviewed / Open Access
• [Journal Article] Efficacy of perioperative chemotherapy for synovial sarcoma: a retrospective analysis of a Nationwide database in Japan (2021)
  • Author(s): Xu Gang、Aiba Hisaki、Yamamoto Norio、Hayashi Katsuhiro、Takeuchi Akihiko、Miwa Shinji、Higuchi Takashi、Abe Kensaku、Taniguchi Yuta、Araki Yoshihiro、Saito Shiro、Yoshimura Kenichi、Murakami Hideki、Tsuchiya Hiroyuki、Kawai Akira
  • Journal Title: BMC Cancer Volume: 21 Issue: 1 Pages: 773-773
  • DOI: 10.1186/s12885-021-08485-1
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Delayed Initiation of Treatment Is Associated With Metastasis of Malignant Bone Tumor (2021)
  • Author(s): ARAKI YOSHIHIRO、YAMAMOTO NORIO、HAYASHI KATSUHIRO、TAKEUCHI AKIHIKO、MIWA SHINJI、IGARASHI KENTARO、HIGUCHI TAKASHI、ABE KENSAKU、TANIGUCHI YUTA、YONEZAWA HIROTAKA、MORINAGA SEI、ASANO YOHEI、TSUCHIYA HIROYUKI
  • Journal Title: Anticancer Research Volume: 41 Issue: 6 Pages: 2993-2999
  • DOI: 10.21873/anticanres.15081
  • Peer Reviewed
• [Journal Article] Osteosarcoma-Derived Small Extracellular Vesicles Enhance Tumor Metastasis and Suppress Osteoclastogenesis by miR-146a-5p (2021)
  • Author(s): Yoshihiro Araki, Hisaki Aiba, Takeshi Yoshida, Norio Yamamoto, Katsuhiro Hayashi, Akihiko Takeuchi, Shinji Miwa, Kentaro Igarashi, Tuan D. Nguyen, Kiyo-aki Ishii, Takayuki Nojima, Satoru Takahashi, Hideki Murakami, Hiroyuki Tsuchiya and Rikinari Hanayama
  • Journal Title: Frontiers in Oncology Volume: 11 Pages: 667109-667109
  • DOI: 10.3389/fonc.2021.667109
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Associations of PD-L1, PD-L2, and HLA class I expression with responses to immunotherapy in patients with advanced sarcoma: post hoc analysis of a phase 1/2 trial (2021)
  • Author(s): Miwa S.、Nojima T.、Alomesen A. A.、Ikeda H.、Yamamoto N.、Nishida H.、Hayashi K.、Takeuchi A.、Igarashi K.、Higuchi T.、Yonezawa H.、Araki Y.、Morinaga S.、Asano Y.、Tsuchiya H.
  • Journal Title: Clinical and Translational Oncology Volume: 23 Issue: 8 Pages: 1620-1629
  • DOI: 10.1007/s12094-021-02559-z
  • Peer Reviewed / Open Access
• [Presentation] Over 7-years follow-up of tumor-devitalized frozen autografts of long bones for the patients with malignant bone tumors (2021)
  • Author(s): Akihiko Takeuchi, Norio Yamamoto, Katsuhiro Hayashi, Shinji Miwa, Kentaro Igarashi, Yuta Taniguchi, Yoshihiro Araki, Hirotaka Yonezawa, Sei Morinaga, Hiroyuki Tsuchiya
  • Organizer: The 13th Asia Pacific Musculoskeletal Tumor Society Meeting
  • Int'l Joint Research
• [Presentation] Randomized placebo-contorolled double blind phase II study of zaltoprofen for patient with diffuse-type and unresectable localized tenosynovial giant cell tumors (2021)
  • Author(s): Akihiko Takeuchi,Makoto Endo, Akira Kawai, Yoshihiro Nishida, Ryu Terauchi, Akihiko Matsumine, Hisaki Aiba, Tomoki Nakamura, Susumu Tandai, Toshifumi Ozaki, Manabu Hoshi, Kenichi Yoshimura, Akihiro Nomura,Toshinori Murayama, Hiroyuki Tsuchiya
  • Organizer: The Connective Tissue Oncology Society Annual Meeting
  • Int'l Joint Research