Molecular understanding of the accumulation of mesenchymal stem cells in vivo by photo-manipulation techniques and its application to bone-related diseases

• Project/Area Number: 17H04399
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Dental engineering/Regenerative dentistry
• Research Institution: Okayama University
• Principal Investigator: Takarada Takeshi 岡山大学, 医歯薬学総合研究科, 教授 (30377428)
• Co-Investigator(Kenkyū-buntansha): 窪木 拓男 岡山大学, 医歯薬学総合研究科, 教授 (00225195) ; 大野 充昭 岡山大学, 医歯薬学総合研究科, 准教授 (60613156) ; 佐藤 守俊 東京大学, 大学院総合文化研究科, 教授 (00323501) ; 戸村 道夫 大阪大谷大学, 薬学部, 教授 (30314321) ; 戸口田 淳也 京都大学, ウイルス・再生医科学研究所, 教授 (40273502) ; 渡辺 亮 京都大学, iPS細胞研究所, 特定拠点助教 (60506765)
• Project Period (FY): 2017-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥16,640,000 (Direct Cost: ¥12,800,000、Indirect Cost: ¥3,840,000); Fiscal Year 2020: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000); Fiscal Year 2019: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000); Fiscal Year 2018: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000); Fiscal Year 2017: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
• Keywords: 光操作 / 遺伝子改変マウス / 間葉系幹細胞 / 光操作技術 / 幹細胞

Outline of Final Research Achievements

In this study, we generated a tTA-dependent photoactivatable Cre-loxP recombinase knock-in mouse model (TRE-PA-Cre mice) using a CRISPR/Cas9 system. These mice were crossed with ROSA26-tdTomato mice (Cre reporter mouse) to visualize DNA recombination as marked by tdTomato expression. We demonstrated that external noninvasive LED blue light illumination allows efficient DNA recombination in the liver of TRE-PA-Cre:ROSA26-tdTomato mice transfected with tTA expression vectors using hydrodynamic tail vein injection. The TRE-PA-Cre mouse established here promises to be useful for optogenetic genome engineering in a noninvasive, spatiotemporal, and cell-type specific manner in vivo.

Academic Significance and Societal Importance of the Research Achievements

従来不可能であった「時間・空間（光照射時/部位）特異的」な精度を持つ生体内遺伝子操作が可能となり、今後、様々な組織・細胞で生物医学研究の新規戦略手法として幅広く使用されることが期待される。

Research Products

• [Int'l Joint Research] University of Rochester Medical Center(米国)
• [Int'l Joint Research] Monash University(オーストラリア)
• [Int'l Joint Research] University of Rochester Medical Center(米国)
• [Journal Article] PRRX1 promotes malignant properties in human osteosarcoma (2021)
  • Author(s): Joko Ryoji、Yamada Daisuke、Nakamura Masahiro、Yoshida Aki、Takihira Shota、Takao Tomoka、Lu Ming、Sato Kohei、Ito Tatsuo、Kunisada Toshiyuki、Nakata Eiji、Ozaki Toshifumi、Takarada Takeshi
  • Journal Title: Translational Oncology Volume: 14 Issue: 1 Pages: 100960-100960
  • DOI: 10.1016/j.tranon.2020.100960
  • NAID: 120007004820
  • Peer Reviewed / Open Access
• [Journal Article] RUNX2 regulates leukemic cell metabolism and chemotaxis in high-risk T cell acute lymphoblastic leukemia (2021)
  • Author(s): Matthijssens Filip、Sharma Nitesh D.、Takarada Takeshi、et al.
  • Journal Title: Journal of Clinical Investigation Volume: 131 Issue: 6
  • DOI: 10.1172/jci141566
  • Peer Reviewed / Int'l Joint Research
• [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] Core Binding Factors are essential for ovulation, luteinization, and female fertility in mice (2020)
  • Author(s): Lee-Thacker Somang、Jeon Hayce、Choi Yohan、Taniuchi Ichiro、Takarada Takeshi、Yoneda Yukio、Ko CheMyong、Jo Misung
  • Journal Title: Scientific Reports Volume: 10 Issue: 1 Pages: 9921-9921
  • DOI: 10.1038/s41598-020-64257-0
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] A RUNX2 stabilization pathway mediates physiologic and pathologic bone formation (2020)
  • Author(s): Kim Jung-Min、Yang Yeon-Suk、Park Kwang Hwan、Ge Xianpeng、Xu Ren、Li Na、Song Minkyung、Chun Hyunho、Bok Seoyeon、Charles Julia F.、Filhol-Cochet Odile、Boldyreff Brigitte、Dinter Teresa、Yu Paul B.、Kon Ning、Gu Wei、Takarada Takeshi、Greenblatt Matthew B.、Shim Jae-Hyuck
  • Journal Title: Nature Communications Volume: 11 Issue: 1 Pages: 2289-2289
  • DOI: 10.1038/s41467-020-16038-6
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Establishment of a tTA-dependent photoactiveatable Cre recombinease knock-in mouse model for optogenetic genome engineering. (2020)
  • Author(s): 9.Takao, T., Hiraoka, Y., Kawabe, K., Yamada, D., Ming, L., Tanaka, K., Sato, M., Takarada, T.
  • Journal Title: Biochem. Biophys. Res. Commun. Volume: 526 Issue: 1 Pages: 213-217
  • DOI: 10.1016/j.bbrc.2020.03.015
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Inhibition of the glutamine transporter SNAT1 confers neuroprotection in mice by modulating the mTOR-autophagy system (2019)
  • Author(s): Yamada Daisuke、Kawabe Kenji、Tosa Ikue、Tsukamoto Shunpei、Nakazato Ryota、Kou Miki、Fujikawa Koichi、Nakamura Saki、Ono Mitsuaki、Oohashi Toshitaka、Kaneko Mari、Go Shioi、Hinoi Eiichi、Yoneda Yukio、Takarada Takeshi
  • Journal Title: Communications Biology Volume: 2 Issue: 1
  • DOI: 10.1038/s42003-019-0582-4
  • Peer Reviewed / Open Access
• [Journal Article] Glutamatergic neurons in the medial prefrontal cortex mediate the formation and retrieval of cocaine-associated memories in mice (2019)
  • Author(s): Zhang Tong、Yanagida Junko、Kamii Hironori、Wada Shintaro、Domoto Masaki、Sasase Hitoki、Deyama Satoshi、Takarada Takeshi、Hinoi Eiichi、Sakimura Kenji、Yamanaka Akihiro、Maejima Takashi、Mieda Michihiro、Sakurai Takeshi、Nishitani Naoya、Nagayasu Kazuki、Kaneko Shuji、Minami Masabumi、Kaneda Katsuyuki
  • Journal Title: Addiction Biology Volume: 印刷中 Issue: 1 Pages: 1-11
  • DOI: 10.1111/adb.12723
  • Peer Reviewed / Open Access
• [Journal Article] Postnatal Runx2 deletion leads to low bone mass and adipocyte accumulation in mice bone tissues (2019)
  • Author(s): Tosa Ikue、Yamada Daisuke、Yasumatsu Misa、Hinoi Eiichi、Ono Mitsuaki、Oohashi Toshitaka、Kuboki Takuo、Takarada Takeshi
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 516 Issue: 4 Pages: 1229-1233
  • DOI: 10.1016/j.bbrc.2019.07.014
  • Peer Reviewed
• [Journal Article] Type IV collagen α6 chain is a regulator of keratin 10 in keratinization of oral mucosal epithelium (2018)
  • Author(s): Komori T, Ono M, Hara ES, Ueda J, Nguyen HTT, Nguyen HT, Yonezawa T, Maeba T, Ono A, Takarada T, Momota R, Maekawa K, Kuboki T, Oohashi T
  • Journal Title: Scientific Reports Volume: 8 Issue: 1 Pages: 2612-2612
  • DOI: 10.1038/s41598-018-21000-0
  • Peer Reviewed / Open Access
• [Journal Article] Core Binding Factor β Expression in Ovarian Granulosa Cells Is Essential for Female Fertility. (2018)
  • Author(s): Lee-Thacker S, Choi Y, Taniuchi I, Takarada T, Yoneda Y, Ko C, Jo M.
  • Journal Title: Endocrinology. Volume: 159 Issue: 5 Pages: 2094-2109
  • DOI: 10.1210/en.2018-00011
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] The MAPK Erk5 is necessary for proper skeletogenesis involving a Smurf-Smad-Sox9 molecular axis. (2018)
  • Author(s): Iezaki T, Fukasawa K, Horie T, Park G, Robinson S, Nakaya M, Fujita H, Onishi Y, Ozaki K, Kanayama T, Hiraiwa M, Kitaguchi Y, Kaneda K, Yoneda Y, Takarada T, Guo XE, Kurose H, Hinoi E.
  • Journal Title: Development Volume: 145 Issue: 14 Pages: 164004-164004
  • DOI: 10.1242/dev.164004
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Radiobrominated benzimidazole-quinoline derivatives as Platelet-derived growth factor receptor beta (PDGFRβ) imaging probes. (2018)
  • Author(s): Effendi N, Mishiro K, Takarada T, Makino A, Yamada D, Kitamura Y, Shiba K, Kiyono Y, Odani A, Ogawa K.
  • Journal Title: Sci Rep. Volume: 8 Issue: 1 Pages: 10369-10369
  • DOI: 10.1038/s41598-018-28529-0
  • NAID: 120007185660
  • Peer Reviewed / Open Access
• [Journal Article] Physiological role of urothelial cancer-associated one long noncoding RNA in human skeletogenic cell differentiation (2017)
  • Author(s): Ishikawa T, Nishida T, Ono M, Takarada T, Nguyen HT, Kurihara S, Furumatsu T, Murase Y, Takigawa M, Oohashi T, Kamioka H, Kubota S
  • Journal Title: J Cell. Physiol. Volume: 233(6) Issue: 6 Pages: 4825-4840
  • DOI: 10.1002/jcp.26285
  • Peer Reviewed / Open Access
• [Journal Article] Bone Resorption Is Regulated by Circadian Clock in Osteoblasts. (2017)
  • Author(s): Takarada T, Xu C, Ochi H, Nakazato R, Yamada D, Nakamura S, Kodama A, Shimba S, Mieda M, Fukasawa K, Ozaki K, Iezaki T, Fujikawa K, Yoneda Y, Numano R, Hida A, Tei H, Takeda S, Hinoi E.
  • Journal Title: J Bone Miner Res. Volume: 32 Issue: 4 Pages: 872-881
  • DOI: 10.1002/jbmr.3053
  • Peer Reviewed
• [Journal Article] Deletion of Runx2 in Articular Chondrocytes Decelerates the Progression of DMM-Induced Osteoarthritis in Adult Mice (2017)
  • Author(s): Liao Lifan、Zhang Shanxing、Gu Jianhong、Takarada Takeshi、Yoneda Yukio、Huang Jian、Zhao Lan、Oh Chun-do、Li Jun、Wang Baoli、Wang Meiqing、Chen Di
  • Journal Title: Scientific Reports Volume: 7 Issue: 1 Pages: 2371-2371
  • DOI: 10.1038/s41598-017-02490-w
  • Peer Reviewed
• [Journal Article] The transcriptional modulator Ifrd1 controls PGC-1α expression under short-term adrenergic stimulation in brown adipocytes (2017)
  • Author(s): Park Gyujin、Horie Tetsuhiro、Kanayama Takashi、Fukasawa Kazuya、Iezaki Takashi、Onishi Yuki、Ozaki Kakeru、Nakamura Yukari、Yoneda Yukio、Takarada Takeshi、Hinoi Eiichi
  • Journal Title: The FEBS Journal Volume: 284 Issue: 5 Pages: 784-795
  • DOI: 10.1111/febs.14019
  • Peer Reviewed
• [Presentation] 組織機能修復・再生・創薬を指向した間葉系幹細胞の幹細胞の階層性/系譜の分子理解 (2018)
  • Author(s): 宝田 剛志
  • Organizer: 第29回創薬・薬理フォーラム岡山
  • Invited
• [Presentation] Dissecting the hierarchy and lineage of mesenchymal stem cells by mouse genetics. (2018)
  • Author(s): Takeshi Takarada
  • Organizer: International Symposium on Immunology and Tissue Regeneration in Okayama / 12th URA International Symposium
• [Presentation] 間葉系幹細胞の階層性・系譜の分子理解と応用 (2017)
  • Author(s): 宝田 剛志
  • Organizer: 第26回硬組織再生生物学会
  • Invited
• [Presentation] 間葉系幹細胞の階層性・系譜の分子理解と骨疾患への応用 (2017)
  • Author(s): 宝田 剛志
  • Organizer: 第54回ネフロンフォーラム
  • Invited
• [Presentation] 骨格形成過程における細胞集団不均一性の理解 (2017)
  • Author(s): 宝田 剛志
  • Organizer: 第5回細胞凝集研究会
  • Invited
• [Presentation] 骨組織再生材料開発を指向した骨格形成過程の階層性・系譜の分子理解 (2017)
  • Author(s): 宝田 剛志
  • Organizer: 日本バイオマテリアル学会
  • Invited
• [Presentation] 生体内での間葉系幹細胞系譜の分子理解と、その応用 (2017)
  • Author(s): 宝田 剛志
  • Organizer: 日本薬学会第138年会
  • Invited