虚血性骨壊死におけるToll-like receptor 4の役割の検討

• Project/Area Number: 19K18471
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 56020:Orthopedics-related
• Research Institution: Nagoya City University
• Principal Investigator: 黒柳 元 名古屋市立大学, 医薬学総合研究院(医学), 講師 (80790831)
• Project Period (FY): 2019-04-01 – 2024-03-31
• Project Status: Granted (Fiscal Year 2022)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2021: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2020: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2019: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
• Keywords: 大腿骨頭壊死症 / 虚血性骨壊死 / Toll-like receptor 4 / TLR4阻害剤 / マウスモデル / 間葉系幹細胞 / Toll-様受容体 / Toll-like reecptor 4 / 大腿骨頭壊死

Outline of Research at the Start

本研究では、生体内におけるTLR4の骨壊死への機能解析のために、TLR4遺伝子ノックアウトマウスに、虚血性骨壊死を誘導し、野生型マウスと比較することで、TLR4欠損が骨壊死にどのように影響するのかを詳細に検討する。また、骨壊死を誘導したマウスモデルに、TLR4阻害剤（TAK242）を投与し、生理食塩水を投与した群と比較することで、TAK242が骨壊死を制御するのかについても詳細に検討する。

Outline of Annual Research Achievements

我が国では小児や成人において大腿骨が腐る（大腿骨頭壊死）は広く知られているが、有効な治療法がないため難病指定を受けている。申請者は全米第1位の小児整形外科の専門病院であるTexas Scottish Rite Hospital for Children（TSRH）と共同研究を開始し、同グループが開発した虚血性骨壊死マウスモデルを用いて、interleukin-6（IL-6）が骨壊死後の骨リモデリングを抑制する分子機序を世界で初めて報告した。また、TSRH研究グループは、骨壊死発生後に壊死に陥った細胞から放出されるdamage associated molecular patterns（DAMPs）がtoll-like receptor 4（TLR4）を介したマクロファージでのinnate immune systemを引き起こし、IL-6などの炎症性サイトカインの放出を促進することが骨壊死の病態に深く関与していると報告している。本研究の目的は骨壊死におけるTLR4の役割のさらなる解明を進め骨壊死の新たな治療法を確立することである。
骨壊死におけるTLR4の機能が解明されれば、骨壊死を制御しうる可能性があり、新たな治療薬開発などの可能性を秘めている。本研究が、現在までの骨壊死の治療体系に大きな変化をもたらす可能性があり、整形外科学の発展に資する重要な研究であると確信している。

Current Status of Research Progress

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

Reason

今年度は、虚血性骨壊死を誘導したマウスへTLR4阻害薬のResatorvidを腹腔内投与し、術後4週の検体を用いたマイクロCTにおいて、骨壊死後の骨量低下にどのように影響するのかを詳細に検討した。その結果、生理食塩水を投与した群と比較して、Resatorvidが骨壊死による骨量低下を抑制する可能性を見出した。

Strategy for Future Research Activity

今後は、生体内におけるTLR4の骨壊死への機能解析のためTLR4遺伝子ノックアウトマウス（B6.129P2-Tlr4tm1Aki/Obs5）に骨壊死を誘導し、野生型マウス（B6）と比較することで、TLR4欠損が骨壊死にどのように影響するのかを検討する。

Research Products

• [Journal Article] Pin and Wire System Fixation for the Treatment of Comminuted Patella Fractures (2023)
  • Author(s): Iwata Hidetoshi、Takada Naoya、Kuroyanagi Gen、Usami Takuya、Sekiya Isato、Murakami Hideki
  • Journal Title: Orthopedics Volume: 1 Issue: 5 Pages: 1-6
  • DOI: 10.3928/01477447-20230310-01
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Fixation of intra-articular calcaneal fractures: A comparative study of the postoperative outcome between HA/PPLA screws and locking plates (2023)
  • Author(s): Usami Takuya、Takada Naoya、Nishida Kazuki、Sakai Hiroaki、Iwata Hidetoshi、Yonezu Hiroki、Sekiya Isato、Nagaya Yuko、Ueki Yoshino、Murakami Hideki、Kuroyanagi Gen
  • Journal Title: Heliyon Volume: 9 Issue: 3 Pages: e14046-e14046
  • DOI: 10.1016/j.heliyon.2023.e14046
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Oncostatin M attenuates tumor necrosis factor-α-induced synthesis of macrophage-colony stimulating factor via suppression of Akt in osteoblasts. (2023)
  • Author(s): Hioki T, Kuroyanagi G, Matsushima-Nishiwaki R, Kozawa O, Tokuda H.
  • Journal Title: Connect. Tissue Res. Volume: 64 Issue: 2 Pages: 139-147
  • DOI: 10.1080/03008207.2022.2109468
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] HSP70 inhibitors upregulate prostaglandin E1-induced synthesis of interleukin-6 in osteoblasts (2022)
  • Author(s): Kuroyanagi Gen、Tachi Junko、Fujita Kazuhiko、Kawabata Tetsu、Sakai Go、Nakashima Daiki、Kim Woo、Tanabe Kumiko、Matsushima-Nishiwaki Rie、Otsuka Takanobu、Iida Hiroki、Kozawa Osamu、Tokuda Haruhiko
  • Journal Title: PLOS ONE Volume: 17 Issue: 12 Pages: e0279134-e0279134
  • DOI: 10.1371/journal.pone.0279134
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Evaluation of lower extremity gait analysis using Kinect V2^? tracking system (2022)
  • Author(s): Usami Takuya、Nishida Kazuki、Iguchi Hirotaka、Okumura Taro、Sakai Hiroaki、Ida Ruido、Horiba Mitsuya、Kashima Shuuto、Sahashi Kento、Asai Hayato、Nagaya Yuko、Murakami Hideki、Ueki Yoshino、Kuroyanagi Gen
  • Journal Title: SICOT-J Volume: 8 Pages: 27-27
  • DOI: 10.1051/sicotj/2022027
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Upregulation of TGF-β-induced HSP27 by HSP90 inhibitors in osteoblasts (2022)
  • Author(s): Kuroyanagi Gen、Tokuda Haruhiko、Fujita Kazuhiko、Kawabata Tetsu、Sakai Go、Kim Woo、Hioki Tomoyuki、Tachi Junko、Matsushima-Nishiwaki Rie、Otsuka Takanobu、Iida Hiroki、Kozawa Osamu
  • Journal Title: BMC Musculoskeletal Disorders Volume: 23 Issue: 1 Pages: 495-495
  • DOI: 10.1186/s12891-022-05419-1
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Clinical and radiographic outcomes of primary total hip arthroplasty with the revelation hip system using density mapping (2022)
  • Author(s): Sakai Hiroaki、Watanabe Nobuyuki、Kuroyanagi Gen、Fukuoka Muneyoshi、Usami Takuya、Ueki Yoshino、Murakami Hideki
  • Journal Title: European Journal of Orthopaedic Surgery and Traumatology Volume: 33 Issue: 2 Pages: 435-440
  • DOI: 10.1007/s00590-022-03210-5
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Attenuation by HSP90 inhibitors of EGF-elicited migration of osteoblasts: involvement of p44/p42 MAP kinase (2022)
  • Author(s): Kuroyanagi Gen、Kawabata Tetsu、Tokuda Haruhiko、Fujita Kazuhiko、Matsushima-Nishiwaki Rie、Sakai Go、Tachi Junko、Hioki Tomoyuki、Kim Woo、Iida Hiroki、Otsuka Takanobu、Kozawa Osamu
  • Journal Title: Connective Tissue Research Volume: 63 Issue: 4 Pages: 359-369
  • DOI: 10.1080/03008207.2021.1939323
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Impact of social support on severity of depressive symptoms by remission status in patients with rheumatoid arthritis (2021)
  • Author(s): Yasuoka Mikako、Kojima Toshihisa、Waguri-Nagaya Yuko、Saito Tami、Takahashi Nobunori、Asai Shuji、Sobue Yasumori、Nishiume Tsuyoshi、Suzuki Mochihito、Mitsui Hiroto、Kawaguchi Yohei、Kuroyanagi Gen、Kamiji Koto、Watanabe Miki、Suzuki Sadao、Kondo Katsunori、Ojima Toshiyuki、Kojima Masayo
  • Journal Title: Modern Rheumatology Volume: - Issue: 3 Pages: 528-533
  • DOI: 10.1093/mr/roab001
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Diagnosis of medial meniscal ramp lesion is difficult by pre-operative magnetic resonance imaging evaluation and needs a methodical arthroscopic exploration (2021)
  • Author(s): Yasuma Sanshiro、Kobayashi Makoto、Kawanishi Yusuke、Fukushima Hiroaki、Kato Jiro、Takenaga Tetsuya、Yoshida Masahito、Kuroyanagi Gen、kawaguchi Yohei、Nagaya Yuko、Murakami Hideki、Nozaki Masahiro
  • Journal Title: Journal of Orthopaedic Science Volume: - Issue: 6 Pages: 1271-1277
  • DOI: 10.1016/j.jos.2021.07.020
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Development of a murine model of ischemic osteonecrosis to study the effects of aging on bone repair (2021)
  • Author(s): Yamaguchi Ryosuke、Kamiya Nobuhiro、Kuroyanagi Gen、Ren Yinshi、Kim Harry K. W.
  • Journal Title: Journal of Orthopaedic Research Volume: - Issue: 12 Pages: 2663-2670
  • DOI: 10.1002/jor.25006
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Banding with lesser trochanter fragment using nonabsorbable tape in trochanteric femoral fractures (2021)
  • Author(s): Usami Takuya、Takada Naoya、Nishida Kazuki、Sakai Hiroaki、Iwata Hidetoshi、Sekiya Isato、Ueki Yoshino、Murakami Hideki、Kuroyanagi Gen
  • Journal Title: SICOT-J Volume: 7 Pages: 33-33
  • DOI: 10.1051/sicotj/2021032
  • Peer Reviewed
• [Journal Article] Olive polyphenols attenuate TNF-α-stimulated M-CSF and IL-6 synthesis in osteoblasts: Suppression of Akt and p44/p42 MAP kinase signaling pathways (2021)
  • Author(s): Hioki Tomoyuki、Tokuda Haruhiko、Kuroyanagi Gen、Kim Woo、Tachi Junko、Matsushima-Nishiwaki Rie、Iida Hiroki、Kozawa Osamu
  • Journal Title: Biomedicine & Pharmacotherapy Volume: 141 Pages: 111816-111816
  • DOI: 10.1016/j.biopha.2021.111816
  • Peer Reviewed
• [Journal Article] Anterolateral ligament reconstruction in addition to primary double-bundle anterior cruciate ligament reconstruction for grade 3 pivot shift improves residual knee instability during surgery (2021)
  • Author(s): Kawanishi Yusuke、Kobayashi Makoto、Yasuma Sanshiro、Fukushima Hiroaki、Kato Jiro、Murase Atsunori、Takenaga Tetsuya、Yoshida Masahito、Kuroyanagi Gen、Kawaguchi Yohei、Nagaya Yuko、Murakami Hideki、Nozaki Masahiro
  • Journal Title: Journal of Experimental Orthopaedics Volume: 8 Issue: 1 Pages: 51-51
  • DOI: 10.1186/s40634-021-00369-4
  • Peer Reviewed
• [Journal Article] Effect of hydroxyapatite tubes on the lag screw intraoperative insertion torque for the treatment of intertrochanteric femoral fractures (2021)
  • Author(s): Iwata Hidetoshi、Takada Naoya、Kuroyanagi Gen、Ikuta Kenji、Usami Takuya、Sekiya Isato、Murakami Hideki
  • Journal Title: Injury Volume: 52 Issue: 11 Pages: 3377-3381
  • DOI: 10.1016/j.injury.2021.07.032
  • Peer Reviewed
• [Journal Article] An Analysis of the Femoral Drilling Angle to Avoid Tunnel Collision during Double-Bundle Anterior Cruciate Ligament and Anterolateral Ligament Reconstruction on the Knee (2021)
  • Author(s): Kawanishi Yusuke、Kobayashi Makoto、Yasuma Sanshiro、Fukushima Hiroaki、Kato Jiro、Murase Atsunori、Takenaga Tetsuya、Yoshida Masahito、Kuroyanagi Gen、Kawaguchi Yohei、Nagaya Yuko、Murakami Hideki、Nozaki Masahiro
  • Journal Title: The Journal of Knee Surgery Volume: - Issue: 05 Pages: 0-0
  • DOI: 10.1055/s-0041-1736196
  • Peer Reviewed
• [Journal Article] HSP22 (HSPB8) positively regulates PGF2α-induced synthesis of interleukin-6 and vascular endothelial growth factor in osteoblasts (2021)
  • Author(s): Kuroyanagi Gen、Sakai Go、Otsuka Takanobu、Yamamoto Naohiro、Fujita Kazuhiko、Kawabata Tetsu、Matsushima-Nishiwaki Rie、Kozawa Osamu、Tokuda Haruhiko
  • Journal Title: Journal of Orthopaedic Surgery and Research Volume: 16 Issue: 1 Pages: 72-72
  • DOI: 10.1186/s13018-021-02209-8
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Resveratrol suppresses insulin-like growth factor I-induced osteoblast migration: attenuation of the p44/p42 MAP kinase pathway (2020)
  • Author(s): Hioki Tomoyuki、Kawabata Tetsu、Sakai Go、Fujita Kazuhiko、Kuroyanagi Gen、Matsushima-Nishiwaki Rie、Kim Woo、Otsuka Takanobu、Iida Hiroki、Tokuda Haruhiko、Kozawa Osamu
  • Journal Title: Bioscience, Biotechnology, and Biochemistry Volume: 84 Issue: 12 Pages: 2428-2439
  • DOI: 10.1080/09168451.2020.1809987
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Tramadol enhances PGF 2α-stimulated osteoprotegerin synthesis in osteoblasts (2020)
  • Author(s): Kim W, Tanabe K, Kuroyanagi G, Matsushima-Nishiwaki R, Fujita K, Kawabata T, Sakai G, Tachi J, Hioki T, Nakashima D, Yamaguchi S, Otsuka T, Tokuda H, Kozawa O, Iida H
  • Journal Title: Heliyon Volume: 24 Issue: 8 Pages: e04779-e04779
  • DOI: 10.1016/j.heliyon.2020.e04779
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Depression, physical function, and disease activity associated with frailty in patients with rheumatoid arthritis (2020)
  • Author(s): Kojima Masayo、Kojima Toshihisa、Waguri-Nagaya Yuko、Takahashi Nobunori、Asai Shuji、Sobue Yasumori、Nishiume Tsuyoshi、Suzuki Mochihito、Mitsui Hiroto、Kawaguchi Yohei、Kuroyanagi Gen、Yasuoka Mikako、Watanabe Miki、Suzuki Sadao、Arai Hidenori
  • Journal Title: Modern Rheumatology Volume: 2 Issue: 5 Pages: 1-8
  • DOI: 10.1080/14397595.2020.1838402
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Preoperative Knee Instability Affects Residual Instability as Evaluated by Quantitative Pivot-Shift Measurements During Double-Bundle ACL Reconstruction (2020)
  • Author(s): Kawanishi Yusuke、Nozaki Masahiro、Kobayashi Makoto、Yasuma Sanshiro、Fukushima Hiroaki、Murase Atsunori、Takenaga Tetsuya、Yoshida Masahito、Kuroyanagi Gen、Kawaguchi Yohei、Nagaya Yuko、Murakami Hideki
  • Journal Title: Orthopaedic Journal of Sports Medicine Volume: 8 Issue: 10 Pages: 1-7
  • DOI: 10.1177/2325967120959020
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Treatment of patellar fractures using bioresorbable forged composites of raw particulate unsintered hydroxyapatite/poly-L-lactide cannulated screws and nonabsorbable sutures (2020)
  • Author(s): Usami Takuya、Takada Naoya、Sakai Hiroaki、Endo Shinichiro、Sekiya Isato、Ueki Yoshino、Murakami Hideki、Kuroyanagi Gen
  • Journal Title: Injury Volume: Dec 29 Issue: 6 Pages: 31045-7
  • DOI: 10.1016/j.injury.2020.12.015
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Incretin accelerates platelet-derived growth factor-BB-induced osteoblast migration via protein kinase A: The upregulation of p38 MAP kinase. (2020)
  • Author(s): Kawabata T, Tokuda H, Kuroyanagi G, Fujita K, Sakai G, Kim W, Matsushima-Nishiwaki R, Iida H, Yata KI, Wang S, Mizoguchi A, Otsuka T, Kozawa O.
  • Journal Title: Sci Rep Volume: 10 Issue: 1 Pages: 2341-2341
  • DOI: 10.1038/s41598-020-59392-7
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Anterolateral ligament reconstruction as an augmented procedure for double-bundle anterior cruciate ligament reconstruction restores rotational stability: Quantitative evaluation of the pivot shift test using an inertial sensor. (2020)
  • Author(s): Yasuma S, Nozaki M, Murase A, Kobayashi M, Kawanishi Y, Fukushima H, Takenaga T, Yoshida M, Kuroyanagi G, Kawaguchi Y, Nagaya Y, Murakami H.
  • Journal Title: Knee Volume: S0968-0160 Issue: 2 Pages: 30053-3
  • DOI: 10.1016/j.knee.2020.02.015
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] IL6 receptor blockade preserves articular cartilage and increases bone volume following ischemic osteonecrosis in immature mice (2019)
  • Author(s): Kamiya N.、Kuroyanagi G.、Aruwajoye O.、Kim H.K.W.
  • Journal Title: Osteoarthritis and Cartilage Volume: 27 Issue: 2 Pages: 326-335
  • DOI: 10.1016/j.joca.2018.10.010
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Enhancement by HSP90 inhibitor of PGD2-stimulated HSP27 induction in osteoblasts: Suppression of SAPK/JNK and p38 MAP kinase. (2019)
  • Author(s): Kim W, Tokuda H, Kawabata T, Fujita K, Sakai G, Nakashima D, Tachi J, Kuroyanagi G, Matsushima-Nishiwaki R, Tanabe K, Otsuka T, Iida H, Kozawa O.
  • Journal Title: Prostaglandins Other Lipid Mediat Volume: 143 Pages: 106327-106327
  • DOI: 10.1016/j.prostaglandins.2019.03.002
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] 虚血性骨壊死におけるインターロイキン6の役割の検討 (2022)
  • Author(s): 黒柳 元, 神谷宣広, 山口亮介, Harry K.W.Kim
  • Organizer: 第259回整形外科集談会東海地方会
• [Presentation] 虚血性骨壊死におけるインターロイキン6の役割の検討 (2021)
  • Author(s): 黒柳元、神谷宣広、山口亮介、Harry K. W. Kim
  • Organizer: 第14回東海関節研究会
• [Presentation] 関節リウマチの過去と未来 関節リウマチに関する鏡視下滑膜切除術の意義 (2021)
  • Author(s): 黒柳元、松井宣夫、坂井宏章、宇佐美琢也、永谷祐子、川口洋平、植木美乃、村上英樹
  • Organizer: 第36回日本臨床リウマチ学会
  • Invited
• [Presentation] Interleukin-6 Deletion Induces Revascularization and New Bone Formation in a Mouse Model of Ischemic Osteonecrosis (2019)
  • Author(s): Kuroyanagi G, Kamiya N, Yamaguchi R, Nagaya Y, Wada I, Kim HKW
  • Organizer: 第92回日本整形外科学会学術総会
• [Presentation] Interleukin-6 Receptor Blocker Increases Revascularization and New Bone Formation in a Murine Model of Ischemic Osteonecrosis (2019)
  • Author(s): Kuroyanagi G, Nagaya Y, Yamaguchi R, Kim HKW
  • Organizer: 第63回日本リウマチ学会総会・学術集会
• [Presentation] パーキンソン病患者の急速破壊型股関節症に対してデュオドーパ導入後人工股関節置換術を行った1例 (2019)
  • Author(s): 黒柳 元、植木 美乃、三井 裕人、堀場 充哉、村上 里奈、青山 公紀、井口 普敬、村上 英樹、佐藤 豊大、松川 則之
  • Organizer: 第45回日本リハビリテーション医学会中部・東海地方会