Development of a therapeutic platform for the treatment of craniosynostosis using exosome-delivered siRNA.

• Project/Area Number: 19K10400
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 57070:Developmental dentistry-related
• Research Institution: Tokyo Medical and Dental University
• Principal Investigator: KOBAYASHI YUKIHO 東京医科歯科大学, 大学院医歯学総合研究科, 助教 (20596233)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2021: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2020: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2019: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: 頭蓋縫合部早期癒合症 / 頭蓋骨縫合癒合症 / Apert症候群 / アペール症候群 / siRNA / Fgfr2 / 頭蓋骨縫合早期癒合症 / エキソソーム / ドラッグデリバリーシステム

Outline of Research at the Start

生体内のあらゆる細胞が分泌し血中を循環するナノ分泌顆粒をエクソソームと呼び、新規ドラッグデリバリーシステム（DDS）の候補とされている。一方、頭蓋骨縫合癒合症を主徴とするアペール症候群は、FGFR2の機能獲得型点突然変異によるアミノ酸置換により発症する。本研究は、アペール症候群モデルマウス点変異特異的siRNAをエクソソームにより輸送し、マウス頭蓋顎顔面縫合発生に対する影響をin vitroおよびin vivoにて解析することを目的とした。得られる結果は、広く先天性疾患を対象としたsiRNA創薬の発展へと波及することが期待される。

Outline of Final Research Achievements

To construct Ex-siRNAFgfr2S252W, exosomes were isolated and purified from mouse serum using an exosome isolation kit, and siRNA specific for the mouse Fgfr2 point mutation was introduced into the exosomes using the sRNA-into-Exome kit. The craniofacial sutures of embryonic wild-type mice and Apert syndrome model mice developing craniosynostosis were harvested at each developmental stage, and osteoblasts were isolated from the tissues and cultured. The mRNA and MAPK pathway factor phosphorylation that were specifically up-regulated and down-regulated in the cranial coronal sutures of Apert disease model mice were analyzed.

Academic Significance and Societal Importance of the Research Achievements

得られる結果は、広く先天性疾患を対象としたsiRNA創薬の発展へと波及することが期待される。

Research Products

• [Journal Article] Three-dimensional analysis of the palatal morphology in growing patients with Apert syndrome and Crouzon syndrome (2022)
  • Author(s): Ogura, K. Kobayashi, Y. Hikita, R. Tsuji, M. Moriyama, K.
  • Journal Title: Congenit Anom (Kyoto) Volume: なし Issue: 4 Pages: 153-160
  • DOI: 10.1111/cga.12470
  • Peer Reviewed / Open Access
• [Journal Article] Expression pattern of transcriptional enhanced associate domain family member 1 (Tead1) in developing mouse molar tooth (2021)
  • Author(s): Niki, Y. Kobayashi, Y. Moriyama, K.
  • Journal Title: Gene Expr Patterns Volume: 40 Pages: 119182-119182
  • DOI: 10.1016/j.gep.2021.119182
  • Peer Reviewed
• [Journal Article] Mkx regulates the orthodontic tooth movement via osteoclast induction (2021)
  • Author(s): Miyazaki, T. Kurimoto, R. Chiba, T. Matsushima, T. Nakamichi, R. Tsutsumi, H. Takada, K. Yagasaki, L. Kato, T. Shishido, K. Kobayashi, Y. Matsumoto, T. Moriyama, K. Asahara, H.
  • Journal Title: J Bone Miner Metab Volume: 39 Issue: 5 Pages: 780-786
  • DOI: 10.1007/s00774-021-01233-2
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Aberrantly activated Wnt/β-catenin pathway co-receptors LRP5 and LRP6 regulate osteoblast differentiation in the developing coronal sutures of an Apert syndrome (Fgfr2 S252W /+ ) mouse model (2021)
  • Author(s): Nay Myo Min Swe, Yukiho Kobayashi, Hiroyuki Kamimoto, Keiji Moriyama.
  • Journal Title: Dev Dyn . Volume: 250 Issue: 3 Pages: 465-476
  • DOI: 10.1002/dvdy.239
  • Peer Reviewed
• [Journal Article] Craniofacial, oral, and cervical morphological characteristics in Japanese patients with Apert syndrome or Crouzon syndrome (2020)
  • Author(s): Kobayashi Y, Ogura K, Hikita R, Tsuji M, Moriyama K
  • Journal Title: Eur J Orthod Volume: ー Issue: 1 Pages: 36-44
  • DOI: 10.1093/ejo/cjaa015
  • Peer Reviewed / Open Access
• [Presentation] Expression pattern of transcriptional enhanced associate domain family member 1 in the developing mouse molar tooth (2021)
  • Author(s): Niki Y, Kobayashi Y, Moriyama K
  • Organizer: EOS 2021 Annual Virtual Conference
  • Int'l Joint Research
• [Presentation] "メカニカルストレスによる歯槽骨リモデリングに及ぼす転写制御因子Mohawk homeobox の作用 Effects of the transcription factor Mohawk homebox on alveolar bone remodeling induced by mechanical stress" (2021)
  • Author(s): "宮崎貴行、栗本遼太、松本力、小林起穂、浅原弘嗣、森山啓司 Miyazaki T, Kurimoto R, Matsumoto T, Kobayashi Y, Asahara H, Moriyama K"
  • Organizer: "第80回日本矯正歯科学会学術大会＆第5回国際会議 The 80th Annual Meeting of the Japanese Orthodontic Society & The 5th International Congress"
• [Presentation] "BWS患者に対する舌縮小術が気道および舌に与える影響：側面頭部X線規格写真を用いた検討 Effects of tongue reduction on the upper airway and tongue of BWS patients " (2021)
  • Author(s): "高橋義充、上園将慶、荒井英絵、鈴木彩子、吉澤英之、舩橋健太、小林起穂、森山啓司 Takahashi Y, Uezono M, Arai H, Suzuki A, Yoshizawa H, Funahashi K, Kobayashi Y, Moriyama K "
  • Organizer: "第80回日本矯正歯科学会学術大会＆第5回国際会議 The 80th Annual Meeting of the Japanese Orthodontic Society & The 5th International Congress"
• [Presentation] Sod3 regulates periodontal tissue remodeling through ROS modulation during orthodontic tooth movement of rat molars (2021)
  • Author(s): Jindarojanakul P, Kobayashi Y, Kamimoto H, Satraha S, Moriyama K
  • Organizer: "第80回日本矯正歯科学会学術大会＆第5回国際会議 The 80th Annual Meeting of the Japanese Orthodontic Society & The 5th International Congress"
• [Presentation] Apert症候群モデルマウスの頭蓋縫合早期癒合症におけるWnt/β-cateninシグナル伝達機構の解析 (2020)
  • Author(s): Kobayashi Y, Min Swe Nay Myo, Kamimoto H, Moriyama K
  • Organizer: 第60回日本先天異常学会学術集会
• [Presentation] Aberrant Wnt/β-catenin Signaling in Coronal Sutures of Apert Mouse Model (2020)
  • Author(s): Kobayashi Y, Min Swe Nay Myo, Kamimoto H, Moriyama K
  • Organizer: "The 9th Internarional Orthodontic Congress The 12th Asian Pacific Orthodontic Conference The 79th Annual Meeting of the Japanese Orthodontic Society"
  • Int'l Joint Research
• [Presentation] Aberrant Wnt/β-catenin Signaling in Coronal Sutures of Apert Mouse Model (2020)
  • Author(s): Min Swe Nay Myo, Kobayashi Yukiho, Kamimoto Hiroyuki, Moriyama Keiji
  • Organizer: 2020 IADR/AADR/CADR General Session (Washington, D.C., USA)
  • Int'l Joint Research
• [Presentation] 磁性制御型リポソームにより標的指向化されたリラキシンはラット正中蓋縫合拡大と骨形成を促進する (2019)
  • Author(s): 紙本裕幸、小林起穂、森山啓司
  • Organizer: 第84回口腔病学会学術大会
• [Presentation] "Aberrant activation of Wnt/β-catenin signaling in the coronal sutures of an Apert syndrome mouse model" (2019)
  • Author(s): Minswe NM, Kobayashi Y, Kamimoto H, Moriyama K
  • Organizer: 第78回日本矯正歯科学会学術大会
• [Presentation] 井上貴裕、庄司あゆみ、小倉健司、狩野桜子、佐川夕季、小林起穂、辻美千子、松本力、森山啓司 (2019)
  • Author(s): "トリーチャー・コリンズ症候群患者の下顎骨における antegonial notchの形態に関する検討 "
  • Organizer: 第78回日本矯正歯科学会学術大会
• [Presentation] "Hippo-Yap/Tazシグナル関連転写因子TEA domain family member 1 (Tead1)のマウス歯胚発生における発現解析" (2019)
  • Author(s): 仁木佑紀、小林起穂、森山啓司
  • Organizer: 第78回日本矯正歯科学会学術大会
• [Presentation] Analysis of dentocraniofacial morphology in patients with achondroplasia (2019)
  • Author(s): Tsuji M, Shoji A, Hirabayashi K, Kobayashi Y, Moriyama K
  • Organizer: 第59回日本先天異常学会・The 13th World Congress of International Cleft Lip and Palate oundation CLEFT 2019 ICPF合同学術集会
  • Int'l Joint Research