Analysis of mechanism of bone fragility and effects of drugs in osteogenesis imperfecta using a graft model for enchondral bone formation

• Project/Area Number: 18H02780
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 52050:Embryonic medicine and pediatrics-related
• Research Institution: Osaka University
• Principal Investigator: Ozono Keiichi 大阪大学, 医学系研究科, 教授 (20270770)
• Co-Investigator(Kenkyū-buntansha): 道上 敏美 地方独立行政法人大阪府立病院機構大阪母子医療センター(研究所), 骨発育疾患研究部門, 部長 (00301804) ; 妻木 範行 京都大学, iPS細胞研究所, 教授 (50303938)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥17,420,000 (Direct Cost: ¥13,400,000、Indirect Cost: ¥4,020,000); Fiscal Year 2020: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2019: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2018: ¥7,800,000 (Direct Cost: ¥6,000,000、Indirect Cost: ¥1,800,000)
• Keywords: 骨形成不全症 / 骨脆弱性 / 疾患特異的iPS細胞 / 脆弱性骨折 / ノックインマウス / 骨石灰化 / 成長 / 成長軟骨帯 / 骨系統疾患 / 骨折 / iPS細胞 / モデルマウス / 遺伝子診断 / 薬剤開発

Outline of Final Research Achievements

We obtained fibroblasts and iPS cells from patients with osteogenesis imperfecta (OI). Collagen type 1 from fibroblasts and osteoblasts differentiated from iPS cells were analyzed by SDS-PAGE and LC/MS. The glycosylation-overmodification of collagen type 1 was found. 4-phenyl butyric acid improved overmodificatin and accumulation of collagen in ER. TGF-b signaling was slightly different between osteoblasts induced from normal and OI patients derived iPS cells. The concentration of sclerostin was related to height and weight in 19 OI patients. Variants responsible for OI was found in 36 out of 47 OI patients. The significant genotype-phenotype correlation was reported. The model mice having p.G810S were generated by the knock-in method and proved to show fragile bone phenotypes.

Academic Significance and Societal Importance of the Research Achievements

まだ、有効な治療法のない重症骨形成不全症の治療薬として、ブフェニールを見出した。独自の骨形成不全症モデルマウスを作成し、ブフェニールの効果を判定できる段階となっている。ブフェニールは、他の疾患に適応が取れており、いわゆるdrug repositioningにより、薬剤開発の面でも有利であると考えられる。

Research Products

• [Journal Article] 4-Phenylbutyric acid enhances the mineralization of osteogenesis imperfecta iPSC-derived osteoblasts (2021)
  • Author(s): Takeyari Shinji、Kubota Takuo、Ohata Yasuhisa、Fujiwara Makoto、Kitaoka Taichi、Taga Yuki、Mizuno Kazunori、Ozono Keiichi
  • Journal Title: Journal of Biological Chemistry Volume: 296 Pages: 100027-100027
  • DOI: 10.1074/jbc.ra120.014709
  • Peer Reviewed / Open Access
• [Journal Article] Comprehensive genetic analyses using targeted next-generation sequencing and genotype-phenotype correlations in 53 Japanese patients with osteogenesis imperfecta (2019)
  • Author(s): Ohata Y., Takeyari S., Nakano Y., Kitaoka T., Nakayama H., Bizaoui V., Yamamoto K., Miyata K., Yamamoto K., Fujiwara M., Kubota T., Michigami T., Yamamoto K., Yamamoto T., Namba N., Ebina K., Yoshikawa H., Ozono K.
  • Journal Title: Osteoporosis International Volume: 30 Issue: 11 Pages: 2333-2342
  • DOI: 10.1007/s00198-019-05076-6
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] 患者由来線維芽細胞、iPS細胞を用いた骨形成不全症の病態解析と4-フェニル酪酸の効果（若手研究奨励賞受賞） (2021)
  • Author(s): 武鑓真司, 大幡泰久, 藤原誠, 北岡太一, 窪田拓生, 多賀裕喜, 水野一乗, 大薗恵一.
  • Organizer: 第94回 日本内分泌学会学術総会
• [Presentation] 4-phenylbutyric acid likely improves the quality of extracellular matrix and promotes mineralization in patients with osteogenesis imperfecta-derived cells. (2020)
  • Author(s): Takeyari S, Ohata Y, Fujiwara M, Kitaoka T, Kubota T, Taga Y, Mizuno K, Ozono K.
  • Organizer: ICCBH VIRTUAL FORUM 2020 Bone Fragility Disorders in Children
  • Int'l Joint Research
• [Presentation] 患者由来線維芽細胞、iPS細胞を用いた骨形成不全症の病態解析と4-フェニル酪酸の効果 (2019)
  • Author(s): 武鑓真司，大幡泰久，窪田拓生，多賀祐喜，水野一乘，大薗惠一
  • Organizer: 第51回 日本結合組織学会学術大会
• [Presentation] Analysis of osteogenesis imperfecta in pathology and the effects of 4-phenylbutyric acid using patient-derived fibroblasts and induced pluripotent stem c (2019)
  • Author(s): Shinji Takeyari, Yasuhisa Ohata, Takuo Kubota, Yuki Taga, Kazunori Mizuno, Keiichi Ozono
  • Organizer: ICCBH 2019
  • Int'l Joint Research
• [Presentation] 患者由来線維芽細胞、iPS細胞を用いた骨形成不全症の病態解析と4-フェニル酪酸の効果 (Young Investigator Award) (2019)
  • Author(s): 武鑓真司, 大幡泰久, 窪田拓生, 多賀祐喜, 水野一乗, 大薗恵一
  • Organizer: 第51回 日本結合組織学会学術大会
• [Presentation] Analysis of osteogenesis imperfecta in pathology and the effects of 4-phenylbutyric acid using patient-derived fibroblasts and induced pluripotent stem cells (New Investigator Award) (2019)
  • Author(s): Takeyari S, Ohata Y, Kubota T, Taga Y, Mizuno K, Ozono K.
  • Organizer: 9th International Conference on Children's Bone Health
  • Int'l Joint Research
• [Presentation] ターゲットエクソーム解析を用いた日本人骨形成不全患者47名の網羅的遺伝子解析と表現型解析 (2018)
  • Author(s): 大幡泰久，武鑓真司，中野由佳子，山本賢一，中山尋文，北岡太一，窪田拓生，道上敏美，山本威久，大薗恵一
  • Organizer: 第36回 日本骨代謝学会学術集会
• [Presentation] Comprehensive Genetic Analysis by Targeted Next Generation Sequencing and Genotype-phenotype Correlation of 47 Japanese Patients with Osteogenesis Imperfecta (2018)
  • Author(s): Ohata Y， Takeyari S， Kitaoka T, Nakayama H, Varoona Bizaoui, Nakano N, Yamamoto K, Miyata K, Yamamoto K, Kubota T, Yamamoto K, Michigami T, Yamamoto T, Ozono K
  • Organizer: ASBMR2018 Auual Meeting
  • Int'l Joint Research
• [Presentation] 患者由来iPS細胞、線維芽細胞を用いた骨形成不全症新規治療薬の探索（最優秀演題賞） (2018)
  • Author(s): 武鑓真司，大幡泰久，窪田拓生，多賀祐喜，水野一乘，大薗恵一
  • Organizer: 第52回 日本小児内分泌学会学術集会
• [Presentation] ターゲットエクソーム解析を用いた日本人骨形成不全患者52名の網羅的遺伝子解析と 表現型解析 (2018)
  • Author(s): 大幡泰久，武鑓真司，中野由佳子，山本賢一，宮田 京，中山尋文 Varoona Bizaoui，北岡太一，窪田拓生，山本勝輔，道上敏美，山本威久，大薗恵一
  • Organizer: 第52回 日本小児内分泌学会学術集会