The clarification of bone formation and absorption mechanism in the bone marrow microenvironment

• Project/Area Number: 19H03842
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 57050:Prosthodontics-related
• Research Institution: Okayama University
• Principal Investigator: Ono Mitsuaki 岡山大学, 医歯薬学域, 准教授 (60613156)
• Co-Investigator(Kenkyū-buntansha): 窪木 拓男 岡山大学, 医歯薬学域, 教授 (00225195) ; 宝田 剛志 岡山大学, 医歯薬学域, 教授 (30377428) ; ハラ エミリオ・サトシ 岡山大学, 医歯薬学域, 研究准教授 (40779443) ; 渡辺 亮 京都大学, 医学研究科, 特定准教授 (60506765) ; 秋山 謙太郎 岡山大学, 大学病院, 講師 (70423291) ; 淺田 騰 岡山大学, 大学病院, 研究准教授 (70803055) ; 枝松 緑 岡山大学, 医歯薬学総合研究科, 助教 (10735343)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥17,550,000 (Direct Cost: ¥13,500,000、Indirect Cost: ¥4,050,000); Fiscal Year 2021: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2020: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2019: ¥8,320,000 (Direct Cost: ¥6,400,000、Indirect Cost: ¥1,920,000)
• Keywords: BMP-2 / 骨形成 / 骨吸収 / 1細胞解析 / 骨髄 / 口腔インプラント / 骨形成・抑制メカニズム

Outline of Research at the Start

BMP-2は，有効な骨再生療法を提供するとして大変期待されている．一方，我々は，骨髄腔内にBMP-2を投与すると逆に，骨形成が抑制され，骨髄腔が拡大するという大変興味深い知見を得た．本申請研究では，①骨髄細胞のシングルセル解析にて，BMP-2投与による骨形成抑制・骨髄腔拡大に関わっている細胞を抽出し，②これら候補細胞が，BMP-2投与下で骨髄ニッチや骨髄腔の維持にどの様にして関わっているのかを解析する．そして，上記の解析より，③骨髄ニッチや骨髄腔の維持に関わりが深い細胞や分子を抽出し，その欠損マウスを用いてBMP-2にて骨形成が誘導可能か検証し，骨髄腔の維持に関わる細胞やその分子を同定する．

Outline of Final Research Achievements

BMP-2 has been widely studied for its potent ability to induce osteoblast differentiation and ectopic bone formation, and is already in clinical use worldwide.　On the other hand, we have shown that BMP-2 inhibits bone formation in the bone marrow.　 However, the mechanism by which BMP-2 suppresses bone formation remains unclear. Therefore, we analyzed the detail of BMP-2 induced bone. Single cell RNA-seq analysis revealed that BMP-2-induced bone formed bone marrow with hematopoietic function. These results suggest that BMP-2 has the ability to regenerate bone marrow as an organ, and may suppress bone formation in the bone marrow in order to reserve space for hematopoiesis.

Academic Significance and Societal Importance of the Research Achievements

BMP-2が臨床応用され，10数年が経過し，その有効性と共に様々な副作用が報告されるようになった．その一つが，BMP-2による骨溶解/吸収作用である．また，Jeppsonらは，BMP-2をラット大腿骨周囲に移植した場合，骨形成を抑制する事を報告しており (Acta Orthop Scand, 1999)，臨床医，基礎研究者共にBMP-2の骨吸収作用に長年疑問を抱いていた．そのため，BMP-2による骨髄での骨吸収メカニズムが明らかになれば，その機能を抑制する中和抗体や活性化することができる因子を同定し，新たな治療法の開発に繋げると考える．

Research Products

• [Journal Article] Preclinical bioequivalence study of E.coli-derived rhBMP-2/β-TCP and autogenous bone in a canine guided-bone regeneration model (2022)
  • Author(s): Nosho Shuji, Ono Mitsuaki, Komori Taishi, Mikai Akihiro, Tosa Ikue, Ishibashi Kei, Tanaka Yukie, Kimura-Ono Aya, Hara Emilio S, Oohashi Toshitaka, Kuboki Takuo
  • Journal Title: Journal of Prosthodontic Research Volume: 66 Issue: 1 Pages: 124-130
  • DOI: 10.2186/jpr.JPR_D_20_00226
  • NAID: 130008140800
  • ISSN: 1883-1958, 1883-9207
  • Peer Reviewed / Open Access
• [Journal Article] Suppression of Bone Necrosis around Tooth Extraction Socket in a MRONJ-like Mouse Model by E-rhBMP-2 Containing Artificial Bone Graft Administration (2021)
  • Author(s): Tanaka Yukie、Aung Kyaw Thu、Ono Mitsuaki、Mikai Akihiro、Dang Anh Tuan、Hara Emilio Satoshi、Tosa Ikue、Ishibashi Kei、Ono-Kimura Aya、Nawachi Kumiko、Kuboki Takuo、Oohashi Toshitaka
  • Journal Title: International Journal of Molecular Sciences Volume: 22 Issue: 23 Pages: 12823-12823
  • DOI: 10.3390/ijms222312823
  • NAID: 120007183234
  • Peer Reviewed / Open Access
• [Journal Article] Imatinib has minimal effects on inflammatory and osteopenic phenotypes in a murine cherubism model (2021)
  • Author(s): Mukai Tomoyuki、Akagi Takahiko、Hiramatsu Asano Sumie、Tosa Ikue、Ono Mitsuaki、Kittaka Mizuho、Ueki Yasuyoshi、Yahagi Ayano、Iseki Masanori、Oohashi Toshitaka、Ishihara Katsuhiko、Morita Yoshitaka
  • Journal Title: Oral Diseases Volume: － Issue: 3 Pages: 0-0
  • DOI: 10.1111/odi.14073
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Distinct Osteogenic Potentials of BMP-2 and FGF-2 in Extramedullary and Medullary Microenvironments (2020)
  • Author(s): Nosho Shuji、Tosa Ikue、Ono Mitsuaki、Hara Emilio Satoshi、Ishibashi Kei、Mikai Akihiro、 Tanaka Yukie、Kimura-Ono Aya、Komori Taishi、Maekawa Kenji、Kuboki Takuo、Oohashi Toshitaka
  • Journal Title: International Journal of Molecular Sciences Volume: 21 Issue: 21 Pages: 7967-7967
  • DOI: 10.3390/ijms21217967
  • NAID: 120006937500
  • Peer Reviewed / Open Access
• [Journal Article] BMP-2/β-TCP Local Delivery for Bone Regeneration in MRONJ-Like Mouse Model (2020)
  • Author(s): Mikai Akihiro、Ono Mitsuaki、Tosa Ikue、Nguyen Ha Thi Thu、Hara Emilio Satoshi、Nosho Shuji、Kimura-Ono Aya、Nawachi Kumiko、Takarada Takeshi、Kuboki Takuo、Oohashi Toshitaka
  • Journal Title: International Journal of Molecular Sciences Volume: 21 Issue: 19 Pages: 7028-7028
  • DOI: 10.3390/ijms21197028
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] DNA Methylation-Based Regulation of Human Bone Marrow-Derived Mesenchymal Stem/Progenitor Cell Chondrogenic Differentiation. (2019)
  • Author(s): Nomura Y, Hara E.S, Yoshioka Y, Nguyen H.T, Nosho S, Komori T, Ishibashi K, Oohashi T, Ono M, Kuboki T.
  • Journal Title: Cells Tissues Organs Volume: 207 Issue: 3-4 Pages: 115-126
  • DOI: 10.1159/000502885
  • Peer Reviewed
• [Presentation] 薬剤関連顎骨壊死様モデルマウスにおけるE-rhBMP-2の治療効果の検討 (2021)
  • Author(s): 三海晃弘，大野充昭，土佐郁恵，納所秋二，大野 彩，縄稚久美子，田仲由希恵，大橋俊孝，窪木拓男
  • Organizer: 公益社団法人日本補綴歯科学会第130回学術大会
• [Presentation] rhBMP-2，rhFGF-2の骨形成能は移植部位に存在する骨髄組織の影響を受ける. (2021)
  • Author(s): 納所秋二，大野充昭，土佐郁恵，石橋 啓，田仲由希恵，大野 彩，大橋俊孝，窪木拓男
  • Organizer: 第51回公益社団法人日本口腔インプラント学会学術大会
• [Presentation] 大腸菌由来rhBMP-2含有人工骨の開発と硬組織再生の未来 (2021)
  • Author(s): 大野充昭
  • Organizer: 日本矯正歯科学会大会
  • Invited
• [Presentation] 骨髄存在・非存在下にてrhBMP-2，rhFGF-2が骨形成に与える影響の検討 (2020)
  • Author(s): 納所秋二，大野充昭，土佐郁恵，三海晃弘，石橋 啓，大野 彩，窪木拓男，大橋俊孝
  • Organizer: 公益社団法人日本補綴歯科学会第129回学術大会
• [Presentation] BMP-2，FGF-2が骨髄内・外における骨形成に及ぼす影響の検討 (2020)
  • Author(s): 納所秋二，大野充昭，Ha Nguyen，三海晃弘, 窪木拓男，大橋俊孝
  • Organizer: 岡山大学次世代研究拠点シンポジウム 2020
• [Presentation] マウス薬剤関連性顎骨壊死モデルにおける BMP-2の骨形成能の検討 (2020)
  • Author(s): 三海晃弘, 大野充昭, 納所秋二, 土佐郁恵, 縄稚久美子, 大野 彩, 窪木拓男, 大橋俊孝
  • Organizer: 岡山大学次世代研究拠点シンポジウム 2020
• [Presentation] 大型動物歯槽骨欠損モデルを用いた大腸菌由来rhBMP-2の骨再生能の検討 (2020)
  • Author(s): 田仲由希恵 ，大野充昭，納所秋二 ，石橋 啓，三海晃弘，土佐郁恵， 笈田育尚 ，大野 彩 ，秋山謙太郎 ，大橋俊孝，入江洋之，窪木拓男
  • Organizer: 岡山大学次世代研究拠点シンポジウム 2020
• [Presentation] scRNA-Seq解析を応用したマウス歯胚発生メカニズムの解明の試み (2020)
  • Author(s): 枝松 緑，小川美帆，井出亮太郎，窪木拓男，大橋俊孝，辻 孝，大野充昭
  • Organizer: 岡山大学次世代研究拠点シンポジウム 2020
• [Presentation] Bone-inducing effects of rhBMP-2 and rhFGF-2 in bone marrow microenvironment (2020)
  • Author(s): Shuji Nosho, Mitsuaki Ono, Akihiro Mikai, Kumiko Nawachi, Aya Kimura-Ono, Emilio S Hara, Takuo Kuboki, Toshitaka Oohashi
  • Organizer: International Association for Dental Research (IADR) 98th General Session and Exhibition and 49th Annual Meeting of the American Association for Dental Research (AADR)
  • Int'l Joint Research
• [Presentation] BMP-2 local administration for MRONJ-mice induced partial bone regeneration (2020)
  • Author(s): Akihiro Mikai, Mitsuaki Ono, Shuji Nosho, Ikue Tosa, Emilio S Hara, Aya Kimura-Ono, Kumiko Nawachi, Takuo Kuboki, Toshitaka Oohashi
  • Organizer: International Association for Dental Research (IADR) 98th General Session and Exhibition and 49th Annual Meeting of the American Association for Dental Research (AADR)
  • Int'l Joint Research
• [Presentation] Type XVIII collagen modulates the keratinization of oral mucosa (2020)
  • Author(s): Ha Thi Thu Nguyen, Mitsuaki Ono, Emilio Satoshi Hara, Taishi Komori, Midori Edamatsu, Tomoko Yonezawa, Aya Kimura-Ono, Kenji Maekawa, Takuo Kuboki, Toshitaka Oohashi
  • Organizer: International Association for Dental Research (IADR) 98th General Session and Exhibition and 49th Annual Meeting of the American Association for Dental Research (AADR)
  • Int'l Joint Research
• [Presentation] 骨髄環境におけるBMP-2誘導性骨形成・骨芽細胞分化抑制メカニズムの解明 (2019)
  • Author(s): 納所秋二，大野充昭，Ha Nguyen，笈田育尚，小盛大志，秋山謙太郎，大橋俊孝, 窪木拓男
  • Organizer: 公益社団法人日本補綴歯科学会第128回学術大会
• [Presentation] A new function of Type XVIII collagen in Basement Membrane of Oral Mucosa: a regulator of Keratinization (2019)
  • Author(s): Ha Thi Thu Nguyen, Mitsuaki Ono, Emilio Satoshi Hara, Taishi Komori, Midori Edamatsu, Tomoko Yonezawa, Aya Kimura-Ono, Kenji Maekawa, Takuo Kuboki, Toshitaka Oohashi
  • Organizer: 11th international conference on proteoglycans in Kanazawa
  • Int'l Joint Research
• [Presentation] Postnatal global deletion of the aggrecan gene impairs endochondral ossification (2019)
  • Author(s): Kei Ishibashi, Hidemi Torihara, Mitsuaki Ono, Emilio Satoshi Hara, Midori Edamatsu, Tomoko Yonezawa, Shuji Nosho, Ikue Tosa, Takeshi Takarada, Takuo Kuboki, Attila Aszodi, Hiroshi Kamioka, Toshitaka Oohashi
  • Organizer: 11th international conference on proteoglycans in Kanazawa
  • Int'l Joint Research