Investigation of epigenetic therapeutic targets for musculoskeletal diseases

• Project/Area Number: 15H04961
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Orthopaedic surgery
• Research Institution: Ehime University
• Principal Investigator: Yuuki Imai 愛媛大学, プロテオサイエンスセンター, 教授 (10423873)
• Co-Investigator(Renkei-kenkyūsha): Inoue Kazuki 愛媛大学, 学術支援センター, 助教 (60623725) ; Sakakibara Iori 愛媛大学, プロテオサイエンスセンター, 助教 (50734662)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥16,380,000 (Direct Cost: ¥12,600,000、Indirect Cost: ¥3,780,000); Fiscal Year 2017: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000); Fiscal Year 2016: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000); Fiscal Year 2015: ¥5,720,000 (Direct Cost: ¥4,400,000、Indirect Cost: ¥1,320,000)
• Keywords: エピジェネティクス / 軟骨 / ゲノムワイド解析 / エピジェネティック制御因子 / Uhrf1 / DNAメチル化 / 軟骨細胞 / 運動器 / 分化 / 遺伝子改変マウス

Outline of Final Research Achievements

The physiological functions of Uhrf1 in skeletal tissues remain unclear. We show that limb mesenchymal cell-specific Uhrf1 conditional knockout mice (cKO) exhibit remarkably shortened long bones that have morphological deformities due to dysregulated chondrocyte differentiation and proliferation. Especially, Mmp13 expression was significantly increased in both mRNA and protein levels in cKO mice. Integrative analyses using RNA-seq and MBD-seq revealed that Uhrf1 deficiency decreased genome-wide DNA methylation and increased gene expression through reduced DNA methylation in the promoter regions of 28 genes, including Hspb1. Hspb1 knockdown in cKO chondrocytes can normalize abnormal expression of genes involved in chondrocyte differentiation such as Mmp13. These results indicate that Uhrf1 governs cell-type specific transcriptional regulation by controlling the genome-wide DNA methylation status and regulating consequent cell differentiation and skeletal maturation.

Research Products

• [Journal Article] DANFIN functions as an inhibitor of transcription factor NF-κB and potentiates the antitumor effect of bortezomib in multiple myeloma (2018)
  • Author(s): Uematsu Atsushi、Kido Kohki、Manabe Erika、Takeda Hiroyuki、Takahashi Hirotaka、Hayashi Minoru、Imai Yuuki、Sawasaki Tatsuya
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 495 Issue: 3 Pages: 2289-2295
  • DOI: 10.1016/j.bbrc.2017.12.142
  • Peer Reviewed / Open Access
• [Journal Article] Uhrf1 is indispensable for normal limb growth by regulating chondrocyte differentiation through specific gene expression (2017)
  • Author(s): Yamashita Michiko、Inoue Kazuki、Saeki Noritaka、Ideta-Otsuka Maky、Yanagihara Yuta、Sawada Yuichiro、Sakakibara Iori、Lee Jiwon、Ichikawa Koichi、Kamei Yoshiaki、Iimura Tadahiro、Igarashi Katsuhide、Takada Yasutsugu、Imai Yuuki
  • Journal Title: Development Volume: 145 Issue: 1 Pages: 157412-157412
  • DOI: 10.1242/dev.157412
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] The HIV co-receptor CCR5 regulates osteoclast function. (2017)
  • Author(s): Lee JW, Hoshino A, Inoue K, Saitou T, Uehara S, Kobayashi Y, Ueha S, Matsushima K, Yamaguchi A, Imai Y, Iimura T.
  • Journal Title: Nat Commun Volume: 8 Issue: 1 Pages: 2226-2226
  • DOI: 10.1038/s41467-017-02368-5
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Genome-wide Association Study of Idiopathic Osteonecrosis of the Femoral Head (2017)
  • Author(s): Sakamoto Y, Yamamoto T, Sugano N, Takahashi D, Watanabe T, Atsumi T, Nakamura J, Hasegawa Y, Akashi K, Narita I, Miyamoto T, Takeuchi T, Ikari K, Amano K, Fujie A, Kubo T, Tada Y, Kaneuji A, Nakamura H, Miyamura T, Kabata T, Yamaji K, Okawa T, Sudo A, Ohzono K, Tanaka Y, Yasunaga Y, Matsuda S, Imai Y
  • Journal Title: Sci Rep. Volume: 7 Issue: 1 Pages: 15035-15035
  • DOI: 10.1038/s41598-017-14778-y
  • NAID: 120006817606
  • Peer Reviewed / Open Access
• [Journal Article] Co-administration of Systemic Zoledronate Promotes Osteogenesis Induced by a Local Co-delivery of Recombinant Human Bone Morphogenetic Protein-2 and Β-tricalcium Phosphate in the Bone Marrow of the Rabbit Femur (2017)
  • Author(s): Mamoto K, Ohta Y, Ichikawa K, Imai Y, Minoda Y, Takaoka K, Nakamura H.
  • Journal Title: J Musculoskelet Res. Volume: 19(3) Issue: 03 Pages: 1650015-1650015
  • DOI: 10.1142/s0218957716500159
  • Peer Reviewed
• [Journal Article] The CUL3-SPOP-DAXX axis is a novel regulator of VEGFR2 expression in vascular endothelial cells (2017)
  • Author(s): Sakaue T, Sakakibara I, Fujisaki A, Uesugi T, Nakashiro K, Hamakawa H, Kubota E, Joh T, Imai Y, Izutani H, Higashiyama S.
  • Journal Title: Sci Rep. Volume: 7 Issue: 1 Pages: 42845-42845
  • DOI: 10.1038/srep42845
  • Peer Reviewed / Open Access
• [Journal Article] Local co-application of zoledronate promotes long-term maintenance of newly formed bone induced by recombinant human bone morphogenetic protein 2. (2016)
  • Author(s): Ichikawa K, Ohta Y, Mamoto K, Mizokawa S, Minoda Y, Imai Y, Takaoka K, Nakamura H.
  • Journal Title: Biochem Biophys Res Commun. Volume: 480 Issue: 3 Pages: 314-320
  • DOI: 10.1016/j.bbrc.2016.10.034
  • Peer Reviewed
• [Journal Article] Identification of Nedd9 as a TGF-β-Smad2/3 target gene involved in RANKL-Induced osteoclastogenesis by comprehensive analysis (2016)
  • Author(s): Omata Y, Nakamura S, Koyama T, Yasui T, Hirose J, Izawa N, Matsumoto T, Imai Y, Seo S, Kurokawa M, Tsutsumi S, Kadono Y, Morimoto C, Aburatani H, Miyamoto T, Tanaka S.
  • Journal Title: PLoS One. Volume: 11 Issue: 6 Pages: e0157992-e0157992
  • DOI: 10.1371/journal.pone.0157992
  • Peer Reviewed / Open Access
• [Journal Article] sFRP4-dependent Wnt signal modulation is critical for bone remodeling during postnatal development and age-related bone loss. (2016)
  • Author(s): Haraguchi R, Kitazawa R, Mori K, Tachibana R, Kiyonari H, Imai Y, Abe T, Kitazawa S.
  • Journal Title: Sci Rep. Volume: 6:25198 Issue: 1 Pages: 1-14
  • DOI: 10.1038/srep25198
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Fatostatin, an SREBP inhibitor, prevented RANKL-induced bone loss by suppression of osteoclast differentiation (2015)
  • Author(s): Inoue K and Imai Y
  • Journal Title: BBA-Molecular Basis of Disease. Volume: 1852 Issue: 11 Pages: 2432-41
  • DOI: 10.1016/j.bbadis.2015.08.018
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] Epigenetic regulator, Uhrf1, positively controls skeletal muscle differentiation (2018)
  • Author(s): Yuichiro Sawada, Tadahiko Kikugawa, Iori Sakakibara, Yusuke Ono, Yuta Yanagihara, Noritaka Saeki, Takashi Saika, Yuuki Imai
  • Organizer: ASBMR 2018
  • Int'l Joint Research
• [Presentation] Uhrf1 deficiency attenuates genome-wide DNA methylation and consequent specific gene expression in chondrocytes in vivo. (2018)
  • Author(s): Michiko Yamashita, Noritaka Saeki, Yuta Yanagihara, Yuichiro Sawada, Jiwon Lee, Tadahiro Iimura, Yuuki Imai
  • Organizer: ECTS 2018
  • Int'l Joint Research / Invited
• [Presentation] エピゲノム制御因子Uhrf1による軟骨特異的遺伝子発現制御 (2017)
  • Author(s): 今井祐記
  • Organizer: 第18回運動器科学研究会
• [Presentation] 骨格筋におけるエピゲノム制御因子Uhrf1の機能解析 (2017)
  • Author(s): 沢田雄一郎,榊原伊織,小野悠介,柳原裕太,佐伯法学,菊川忠彦,雑賀隆史,今井祐記
  • Organizer: 第3回日本筋学会学術集会
• [Presentation] エピゲノム制御因子Uhrf1によるDNAメチル化制御を介した軟骨分化・骨格形成 (2017)
  • Author(s): 山下美智子,井上和樹,佐伯法学,榊原伊織,李智媛,大塚まき,亀井義明,五十嵐勝秀,高田泰次,飯村忠浩,今井祐記
  • Organizer: 第35回日本骨代謝学会学術集会
• [Presentation] Epigenetic Regulator, Uhrf1, Controls not Only Cell Proliferation but Also Chondrocyte Differentiation During Limb Development. (2016)
  • Author(s): Michiko Yamashita, Kazuki Inoue, Iori Sakakibara, Yuuki Imai
  • Organizer: The American Society for Bone and Mineral Research 2016 Annual Meeting
  • Place of Presentation: Atlanta, Georgia, USA
  • Year and Date: 2016-09-16
  • Int'l Joint Research
• [Presentation] エピゲノム制御因子Uhrf1は軟骨分化・骨格形成を制御する (2016)
  • Author(s): 山下美智子、今井祐記
  • Organizer: 第34回日本骨代謝学会学術集会
  • Place of Presentation: 大阪
  • Year and Date: 2016-07-20
• [Presentation] Epigenetic regulator, Uhrf1, is a positive regulator in chondrocyte differentiation. (2016)
  • Author(s): Michiko Yamashita, Kazuki Inoue, Iori Sakakibara, Akari Murakami, Yoshiaki Kamei, Yuuki Imai
  • Organizer: 第13回国際人類遺伝学会
  • Place of Presentation: 京都
  • Year and Date: 2016-04-03
  • Int'l Joint Research
• [Presentation] Epigenetic regulator, Uhrf1, is a positive regulator in chondrocyte differentiation (2016)
  • Author(s): Michiko Yamashita,Kazuki Inoue,Iori Sakakibara,Akari Murakami,Yoshiaki Kamei,Yuuki Imai
  • Organizer: 第13回国際人類遺伝学会
  • Place of Presentation: 国立京都国際会館（京都府京都市）
  • Year and Date: 2016-04-03
  • Int'l Joint Research
• [Presentation] Chromatin remodeling during osteoclast differentiation determined by DNase-seq. (2015)
  • Author(s): Yuuki Imai,Kazuki Inoue
  • Organizer: 第38回日本分子生物学会年会、第88回日本生化学会大会合同大会
  • Place of Presentation: 神戸ポートアイランド（兵庫県神戸市）
  • Year and Date: 2015-12-01
  • Invited
• [Presentation] DNase-seqによる破骨細胞分化を制御する新規転写因子の同定 (2015)
  • Author(s): 今井祐記
  • Organizer: 第30回日本整形外科学会基礎学術集会
  • Place of Presentation: 富山市民プラザ（富山県富山市）
  • Year and Date: 2015-10-22
  • Invited
• [Presentation] 骨粗鬆症の病態生理と治療標的探索の試み (2015)
  • Author(s): 今井祐記
  • Organizer: 第25回日本病態生理学会
  • Place of Presentation: 愛媛大学南加記念ホール（愛媛県松山市）
  • Year and Date: 2015-07-31
  • Invited
• [Presentation] Big data解析を用いた細胞分化制御因子の同定 (2015)
  • Author(s): 今井祐記
  • Organizer: 第33回日本骨代謝学会学術集会
  • Place of Presentation: 京王プラザホテル（東京都新宿区）
  • Year and Date: 2015-07-23
  • Invited
• [Presentation] DNase-seqによる破骨細胞分化を制御する新規転写因子の同定 (2015)
  • Author(s): 井上和樹、今井祐記
  • Organizer: 第1回日本骨免疫学会
  • Place of Presentation: ホテルブリーズベイマリーナ（沖縄県宮古島市）
  • Year and Date: 2015-06-30
• [Presentation] Inhibition of a cholesterol regulator, Srebp2, prevents RANKL-induced bone loss (2015)
  • Author(s): Kazuki Inoue, Yuuki Imai
  • Organizer: 4th Joint Meeting ECTS and IBMS
  • Place of Presentation: Rotterdam, Netherlands
  • Year and Date: 2015-04-25
  • Int'l Joint Research
• [Remarks] 愛媛大学プロテオサイエンスセンター
  • URL: http://www.pros.ehime-u.ac.jp/
• [Remarks] 愛媛大学プロテオサイエンスセンター病態生理解析部門
  • URL: https://www.m.ehime-u.ac.jp/school/imailab/
• [Remarks] 愛媛大学プロテオサイエンスセンター病態生理解析部門ホームページ
  • URL: http://www.m.ehime-u.ac.jp/school/imailab
• [Remarks] 愛媛大学プロテオサイエンスセンターホームページ
  • URL: http://www.pros.ehime-u.ac.jp