Comparative Analysis of Epigenomic Dynamics in Bone Development and Repair and its Application to Bone Regeneration

• Project/Area Number: 20H03885
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 57060:Surgical dentistry-related
• Research Institution: The University of Tokyo
• Principal Investigator: Hojo Hironori 東京大学, 大学院医学系研究科(医学部), 准教授 (80788422)
• Co-Investigator(Kenkyū-buntansha): 大庭 伸介 大阪大学, 大学院歯学研究科, 教授 (20466733)
• Project Period (FY): 2020-04-01 – 2024-03-31
• Project Status: Completed (Fiscal Year 2023)
• Budget Amount: ¥17,550,000 (Direct Cost: ¥13,500,000、Indirect Cost: ¥4,050,000); Fiscal Year 2023: ¥3,380,000 (Direct Cost: ¥2,600,000、Indirect Cost: ¥780,000); Fiscal Year 2022: ¥3,380,000 (Direct Cost: ¥2,600,000、Indirect Cost: ¥780,000); Fiscal Year 2021: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2020: ¥5,720,000 (Direct Cost: ¥4,400,000、Indirect Cost: ¥1,320,000)
• Keywords: 骨再生 / １細胞解析 / シグナルネットワーク / 骨修復 / 一細胞解析 / 遺伝子発現機構 / 細胞系譜 / 遺伝子発現制御 / 骨芽細胞 / 細胞系譜実験 / エピゲノム

Outline of Research at the Start

骨組織は一定の自己修復能を有し、小規模骨欠損に対しては自然治癒するのに対し、大規模骨欠損は不可逆的な骨喪失を引き起こし、生活の質を著しく低下させる。本研究では「骨はどのように治るのか？」という骨再生研究における根源的な問いに対して、マウス遺伝学と一細胞解析技術を統合することで、骨欠損部位での幹細胞の振る舞いと細胞分化機構を解明する。さらに、本研究で明らかになった骨修復機構と骨発生機構との比較解析により骨再生性シグナルを同定し、組織再生への応用を検討する。これにより、組織損傷部位においてde novoで起こる生体反応の理解に基づく、新しい骨再生医療戦略の研究基盤の確立を目指す。

Outline of Final Research Achievements

In this study, we genetically labeled skeletal progenitors and their daughter cells by using cell lineage analysis and visualized the behavior of skeletal progenitors at bone defect sites. Single-cell analysis revealed that skeletal progenitors exhibited a bidirectional differentiation to adipocytes as well as osteoblasts at the site of bone repair. Ligand-receptor analysis further identified candidate of signaling factors involved in this directional differentiation. Among them, CCL9 inhibitor had a positive impact on bone regeneration in vivo. These findings provide an insight into the regulatory mechanism of skeletal progenitor cells locally at the site of bone repair.

Academic Significance and Societal Importance of the Research Achievements

骨再生の確立のためには、骨に存在する骨格系前駆細胞の役割を理解し、その制御方法を明らかにすることが重要である。本研究では、骨欠損部位に存在する骨格系細胞を１細胞レベルで詳細に解析することで、その性質や骨再生への寄与のメカニズムを明らかにした。さらに、局所ではたらくシグナルネットワークの一端を解明し、新規骨再生性薬剤の候補を得た。今後、さらなる解析により、骨格系前駆細胞の制御方法を理解し、その活性を操作することができれば、新たな骨再生戦略の確立に寄与することが期待される。

Research Products

• [Int'l Joint Research] 南カリフォルニア大学(米国)
• [Journal Article] Injectable phase-separated tetra-armed poly(ethylene glycol) hydrogel scaffold allows sustained release of growth factors to enhance the repair of critical bone defects (2024)
  • Author(s): Nepal Shant、Si Jinyan、Ishikawa Shohei、Nishikawa Masaki、Sakai Yasuyuki、Akimoto Aya M.、Okada Hiroyuki、Ohba Shinsuke、Chung Ung-il、Sakai Takamasa、Hojo Hironori
  • Journal Title: Regenerative Therapy Volume: 25 Pages: 24-34
  • DOI: 10.1016/j.reth.2023.11.008
  • Peer Reviewed / Open Access
• [Journal Article] Stem cell-based modeling and single-cell multiomics reveal gene-regulatory mechanisms underlying human skeletal development (2023)
  • Author(s): Tani Shoichiro, Okada Hiroyuki, Onodera Shoko, Chijimatsu Ryota, Seki Masahide, Suzuki Yutaka, Xin Xiaonan, Rowe David W., Saito Taku, Tanaka Sakae, Chung Ung-il, Ohba Shinsuke, Hojo Hironori
  • Journal Title: Cell Reports Volume: 42 Issue: 4 Pages: 112276-112276
  • DOI: 10.1016/j.celrep.2023.112276
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Runt-related Transcription Factors and Gene Regulatory Mechanisms in Skeletal Development and Diseases (2023)
  • Author(s): Hojo Hironori、Ohba Shinsuke
  • Journal Title: Current Osteoporosis Reports Volume: 21 Issue: 5 Pages: 485-492
  • DOI: 10.1007/s11914-023-00808-4
  • Peer Reviewed
• [Journal Article] Modeling of intramembranous ossification using human pluripotent stem cell-derived paraxial mesoderm derivatives. (2023)
  • Author(s): Ikeda Y., Tani S., Moriishi T., Kuroda A., Matsuo Y., Saeki N., Inui-Yamamoto C., Abe M., Maeda T., Rowe DW., Chung UI., Hojo H., Matsushita Y., Sawase T., Ohba S.
  • Journal Title: Regenerative Therapy Volume: 24 Pages: 536-546
  • DOI: 10.1016/j.reth.2023.09.017
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Percolation-induced gel-gel phase separation in a dilute polymer network (2023)
  • Author(s): Ishikawa Shohei、Iwanaga Yasuhide、Uneyama Takashi、Li Xiang、Hojo Hironori、Fujinaga Ikuo、Katashima Takuya、Saito Taku、Okada Yasushi、Chung Ung-il、Sakumichi Naoyuki、Sakai Takamasa
  • Journal Title: Nature Materials Volume: 22 Issue: 12 Pages: 1564-1570
  • DOI: 10.1038/s41563-023-01712-z
  • Peer Reviewed / Open Access
• [Journal Article] Single-cell RNA sequencing unravels heterogeneity of skeletal progenitors and cell-cell interactions underlying the bone repair process (2022)
  • Author(s): Nakayama Mika、Okada Hiroyuki、Seki Masahide、Suzuki Yutaka、Chung Ung-il、Ohba Shinsuke、Hojo Hironori
  • Journal Title: Regenerative Therapy Volume: 21 Pages: 9-18
  • DOI: 10.1016/j.reth.2022.05.001
  • Peer Reviewed / Open Access
• [Journal Article] Runx2 regulates chromatin accessibility to direct the osteoblast program at neonatal stages (2022)
  • Author(s): Hojo Hironori, Saito Taku, He Xinjun, Guo Qiuyu, Onodera Shoko, Azuma Toshifumi, Koebis Michinori, Nakao Kazuki, Aiba Atsu, Seki Masahide, Suzuki Yutaka, Okada Hiroyuki, Tanaka Sakae, Chung Ung-il, McMahon Andrew P., Ohba Shinsuke
  • Journal Title: Cell Reports Volume: 40 Issue: 10 Pages: 111315-111315
  • DOI: 10.1016/j.celrep.2022.111315
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Sp7 Action in the Skeleton: Its Mode of Action, Functions, and Relevance to Skeletal Diseases (2022)
  • Author(s): Hojo Hironori、Ohba Shinsuke
  • Journal Title: International Journal of Molecular Sciences Volume: 23 Issue: 10 Pages: 5647-5647
  • DOI: 10.3390/ijms23105647
  • Peer Reviewed / Open Access
• [Journal Article] Conditional inactivation of the L‐type amino acid transporter LAT1 in chondrocytes models idiopathic scoliosis in mice (2022)
  • Author(s): Iwahashi Sayuki、Lyu Jiajun、Tokumura Kazuya、Osumi Ryoma、Hiraiwa Manami、Kubo Takuya、Horie Tetsuhiro、Demura Satoru、Kawakami Noriaki、Saito Taku、Park Gyujin、Fukasawa Kazuya、Iezaki Takashi、Suzuki Akane、Tomizawa Akane、Ochi Hiroki、Hojo Hironori、Ohba Shinsuke、Hinoi Eiichi
  • Journal Title: Journal of Cellular Physiology Volume: 237 Issue: 11 Pages: 4292-4302
  • DOI: 10.1002/jcp.30883
  • Peer Reviewed / Open Access
• [Journal Article] Runx2 and Runx3 differentially regulate articular chondrocytes during surgically induced osteoarthritis development (2022)
  • Author(s): Nagata K, Hojo H, Chang SH, Okada H, Chijimatsu R, Omata Y, Mori D, Makii Y, Kawata M, Kaneko T, Iwanaga Y, Nakamoto H, Maenohara Y, Tachibana N, Ishikura H, Higuchi J, Taniguchi Y, Ohba S, Chung Ul, Tanaka S, Saito T
  • Journal Title: Nature Communications Volume: 13 Issue: 1 Pages: 6187-6204
  • DOI: 10.1038/s41467-022-33744-5
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Involvement of activator protein-1 family members in β-catenin and p300 association on the genome of PANC-1 cells (2022)
  • Author(s): Doi Tomomitsu、Hojo Hironori、Ohba Shinsuke、Obayashi Kunie、Endo Motoyoshi、Ishizaki Toshimasa、Katoh Akira、Kouji Hiroyuki
  • Journal Title: Heliyon Volume: 8 Issue: 2 Pages: e08890-e08890
  • DOI: 10.1016/j.heliyon.2022.e08890
  • Peer Reviewed / Open Access
• [Journal Article] A neomorphic variant in SP7 alters sequence specificity and causes a high-turnover bone disorder (2022)
  • Author(s): Lui Julian C.、Raimann Adalbert、Hojo Hironori、Dong Lijin、Roschger Paul、Kikani Bijal、Wintergerst Uwe、Fratzl-Zelman Nadja、Jee Youn Hee、Haeusler Gabriele、Baron Jeffrey
  • Journal Title: Nature Communications Volume: 13 Issue: 1 Pages: 700-700
  • DOI: 10.1038/s41467-022-28318-4
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Control of osteocyte dendrite formation by Sp7 and its target gene osteocrin (2021)
  • Author(s): Wang JS, Kamath T, Mazur CM, Mirzamohammadi F, Rotter D, Hojo H, Castro CD, Tokavanich N, Patel R, Govea N, Enishi T, Wu Y, da Silva Martins J, Bruce M, Brooks DJ, Bouxsein ML, Tokarz D, Lin CP, Abdul A, Macosko EZ, Fiscaletti M, Munns CF, Ryder P, Kost-Alimova M, Byrne P, Cimini B, Fujiwara M, Kronenberg HM, Wein MN.
  • Journal Title: Nature Communications Volume: 12 Issue: 1 Pages: 6271-6271
  • DOI: 10.1038/s41467-021-26571-7
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Induction and expansion of human PRRX1+ limb-bud-like mesenchymal cells from pluripotent stem cells (2021)
  • Author(s): Yamada Daisuke、Nakamura Masahiro、Takao Tomoka、Takihira Shota、Yoshida Aki、Kawai Shunsuke、Miura Akihiro、Ming Lu、Yoshitomi Hiroyuki、Gozu Mai、Okamoto Kumi、Hojo Hironori、Kusaka Naoyuki、Iwai Ryosuke、Nakata Eiji、Ozaki Toshifumi、Toguchida Junya、Takarada Takeshi
  • Journal Title: Nature Biomedical Engineering Volume: 5 Issue: 8 Pages: 926-940
  • DOI: 10.1038/s41551-021-00778-x
  • Peer Reviewed
• [Journal Article] Mesenchymal stromal cells in the bone marrow niche consist of multi-populations with distinct transcriptional and epigenetic properties (2021)
  • Author(s): Kanazawa Sanshiro、Okada Hiroyuki、Hojo Hironori、Ohba Shinsuke、Iwata Junichi、Komura Makoto、Hikita Atsuhiko、Hoshi Kazuto
  • Journal Title: Scientific Reports Volume: 11 Issue: 1 Pages: 15811-15811
  • DOI: 10.1038/s41598-021-94186-5
  • Peer Reviewed / Open Access
• [Journal Article] The Progress of Stem Cell Technology for Skeletal Regeneration (2021)
  • Author(s): Tani Shoichiro、Okada Hiroyuki、Chung Ung-il、Ohba Shinsuke、Hojo Hironori
  • Journal Title: International Journal of Molecular Sciences Volume: 22 Issue: 3 Pages: 1404-1404
  • DOI: 10.3390/ijms22031404
  • Peer Reviewed / Open Access
• [Journal Article] Understanding paraxial mesoderm development and sclerotome specification for skeletal repair (2020)
  • Author(s): Tani Shoichiro、Chung Ung-il、Ohba Shinsuke、Hojo Hironori
  • Journal Title: Experimental & Molecular Medicine Volume: 52 Issue: 8 Pages: 1166-1177
  • DOI: 10.1038/s12276-020-0482-1
  • Peer Reviewed / Open Access
• [Journal Article] Hedgehog Activation Regulates Human Osteoblastogenesis (2020)
  • Author(s): Onodera Shoko、Saito Akiko、Hojo Hironori、Nakamura Takashi、Zujur Denise、Watanabe Katsuhito、Morita Nana、Hasegawa Daigo、Masaki Hideki、Nakauchi Hiromitsu、Nomura Takeshi、Shibahara Takahiko、Yamaguchi Akira、Chung Ung-il、Azuma Toshifumi、Ohba Shinsuke
  • Journal Title: Stem Cell Reports Volume: 15 Issue: 1 Pages: 125-139
  • DOI: 10.1016/j.stemcr.2020.05.008
  • Peer Reviewed / Open Access
• [Journal Article] Gene regulatory landscape in osteoblast differentiation (2020)
  • Author(s): Hojo Hironori、Ohba Shinsuke
  • Journal Title: Bone Volume: 137 Pages: 115458-115458
  • DOI: 10.1016/j.bone.2020.115458
  • NAID: 120006988169
  • Peer Reviewed
• [Presentation] Injectable phase-separated tetra-armed polyethylene glycol (PEG) hydrogel scaffold allows sustained release of signaling factors to enhance the repair of cranial bone defects (2023)
  • Author(s): Shant Nepal, Jinyan Si, Shohei Ishikawa, Hiroyuki Okada, Shinsuke Ohba, Ung-il Chung, Takamasa Sakai, Hironori Hojo
  • Organizer: ASBMR 2023
  • Int'l Joint Research
• [Presentation] Emerging gene regulatory networks in skeletal development in the mouse and human models (2023)
  • Author(s): 北條 宏徳
  • Organizer: 第41回 日本骨代謝学会
  • Invited
• [Presentation] Modeling of skeletal development using human pluripotent stem cells and exploring the gene regulatory networks (2023)
  • Author(s): Hironori Hojo
  • Organizer: The Korean Society for Biotechnology and Bioengineering (KSBB) 2023
  • Int'l Joint Research / Invited
• [Presentation] Exploring Gene Regulatory Networks (GRNs) in skeletal development using human Pluripotent Stem Cells (PSCs) and future perspective for clinical applications (2023)
  • Author(s): Hironori Hojo
  • Organizer: International Congress of Osteoporosis 2023
  • Int'l Joint Research / Invited
• [Presentation] Elucidating gene regulatory mechanisms underlying human skeletal development using human pluripotent stem cell-derived bone tissuesand single-cell multiome analysis (2022)
  • Author(s): Shoichiro Tani, Hiroyuki Okada, Masahide Seki, Yutaka Suzuki, Taku Saito, Sakae Tanaka, Ung-il Chung, Hironori Hojo and Shinsuke Ohba
  • Organizer: ECTS 2022
  • Int'l Joint Research
• [Presentation] Understanding mechanisms underlying cell fate specification of dental mesenchymal cells in the developing tooth germ through single-cell transcriptome (2022)
  • Author(s): Dahlia Eldeeb, Hironori Hojo, Hiroyuki Okada, Yutaka Suzuki, Masahide Seki, Junichi Tanaka, Shinsuke Ohba, Ung-il Chung
  • Organizer: MBSJ 2022
• [Presentation] Understanding a Mechanism Underlying Bone Repair Process by Combinational Analysis of Lineage Tracing and Single-Cell RNA Sequencing (2021)
  • Author(s): Mika Nakayama, Hiroyuki Okada, Masahide Seki, Yutaka Suzuki, Ung-il Chung, Shinsuke Ohba, Hironori Hojo
  • Organizer: ISSCR 2021
• [Presentation] 骨格形成における遺伝子制御ランドスケープの解明 (2021)
  • Author(s): 北條宏徳
  • Organizer: 17th Meeting of Bone Biology Forum
  • Invited
• [Presentation] Understanding a mechanism underlying bone repair process and application for bone regeneration (2021)
  • Author(s): Hironori Hojo
  • Organizer: SICEM 2021
  • Int'l Joint Research / Invited
• [Presentation] Runx2は骨格発生においてクロマチンランドスケープを規定する (2020)
  • Author(s): 北條宏徳、齋藤琢、小野寺晶子、東俊文、鄭雄一、大庭伸介
  • Organizer: 第38回日本骨代謝学会
• [Presentation] Runx2 defines the osteoblast chromatin landscape in skeletal development (2020)
  • Author(s): Hojo H, Saito T, Onodera S, Azuma T, Chung UI, Seki M, Suzuki Y, McMahon AP, Ohba S
  • Organizer: American Society for Bone and Mineral Research
  • Int'l Joint Research
• [Remarks] ヒト軟骨内骨化の再現とシングルセルマルチオーム解析を用いた転写制御ネットワークの探求
  • URL: https://www.m.u-tokyo.ac.jp/news/press.html
• [Remarks] 骨形成に必須の転写因子Runx2によるゲノムDNAの制御機構が明らかに
  • URL: https://www.m.u-tokyo.ac.jp/news/press.htmlv