Direct reprogramming of fibroblasts to hemetopoietic stem cells

• Project/Area Number: 23659202
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Experimental pathology
• Research Institution: Kyoto University
• Principal Investigator: ITOH Katsuhiko 京都大学, 医学(系)研究科(研究院), 准教授 (90281097)
• Co-Investigator(Kenkyū-buntansha): MIYACHI Hitoshi 京都大学, ウイルス研究所, 技術専門職員 (90599200)
• Project Period (FY): 2011 – 2013
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2013: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000); Fiscal Year 2012: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2011: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: 再生医学 / 造血幹細胞検出法 / リプログラミング / 造血幹細胞

Research Abstract

We have done following experiments; (1) We generated SCL-reporter mouse strains and have screened "candidates genes" using fibroblasts derived from these mouse strains. (2) Although the transduction of a certain combination of "candidates genes" seemed to induce EGFP expression in a small population of transduced cells, these cells could not reconstitute hematopoiesis in recipient mice. (3) We have generated the mutant mouse strains harboring the mutations in the c-kit locus by Crispr and examined the hematopoietic reconstitution ability of the cells described in (2). These cells could not reconstitute hematopoiesis in recipient mice.
Thus, we have concluded that our "candidates genes" could not reprogram fibroblasts to hematopoietic stem cells.

Research Products

• [Journal Article] The BALB/c-specific polymorphic SIRPA enhances its affinity for human CD47, inhibiting phagocytosis against human cells to promote xenogenic engraftment (2014)
  • Author(s): Iwamoto C, Takenaka K, Urata S, Yamauchi T, Shima T, Kuriyama T, Daitoku S, Saito Y, Miyamoto T, Iwasaki H, Kitabayashi I. Itoh K, Kishimoto J, Kohda D, Matozaki T and Akashi K
  • Journal Title: Exp. Hematol Volume: 42 Pages: 163-171
• [Journal Article] The BALB/c-specific polymorphic SIRPA enhances its affinity for human CD47, inhibiting phagocytosis against human cells to promote xenogeneic engraftment. (2014)
  • Author(s): Iwamoto C
  • Journal Title: Exp Hematol Volume: 42 Issue: 3 Pages: 163-171
  • DOI: 10.1016/j.exphem.2013.11.005
  • Peer Reviewed
• [Journal Article] Overexpression of gankyrin in mouse hepatocytes induces hemangioma by suppressing factor inhibiting hypoxia-inducible factor-1 (FIH-1) and activating hypoxia-inducible factor-1 (2013)
  • Author(s): Liu Y, Higashitsuji H, Higashitsuji H, Itoh K, Sakurai T, Koike K, Hirota K, Fukumoto M and Fujit J
  • Journal Title: Biochem. Biophys. Res. Commun Volume: 432 Pages: 22-27
• [Journal Article] Hypothermia Protects against Fulminant Hepatitis in Mice by Reducing Reactive Oxygen Species Production (2013)
  • Author(s): Sakurai T, Kudo M, Watanabe T, Itoh K, Higashitsuji H, Arizumi T, Inoue T, Hagiwara S, Ueshima K, Nishida N, Fukumoto M and Fujita J
  • Journal Title: Dig. Dis. Volume: 31 Pages: 440-446
• [Journal Article] Oscillatory control of factors determining multipotency and fate in mouse neural progenitors (2013)
  • Author(s): Imayoshi, I., Isomura, A., Harima, Y., Kawaguchi, K., Kori, H., Miyachi, H., Fujiwara, T., Ishidate, F., and Kageyama, R
  • Journal Title: Science Volume: 342 Pages: 1203-1208
  • NAID: 120005343917
• [Journal Article] JMJD1C, a JmjC Domain-Containing Protein, Is Required for Long-Term Maintenance of Male Germ Cells in Mice (2013)
  • Author(s): Kuroki, S., Akiyoshi, M., Tokura, M., Miyachi, H., Nakai, Y., Kimura, H., Shinkai, Y., and Tachibana, M
  • Journal Title: Biol. Reprod
• [Journal Article] Epigenetic regulation of mouse sex determination by the histone demethylase Jmjd1a (2013)
  • Author(s): Kuroki, S., Matoba, S., Akiyoshi, M., Matsumura, Y., Miyachi, H., Mise, N., Abe, K., Ogura, A., Wilhelm, D., Koopman, P., Nozaki, M., Kanai, Y., Shinkai, Y., and Tachibana, M
  • Journal Title: Science Volume: 341 Pages: 1106-1109
  • NAID: 120005323045
• [Journal Article] Interleukin-7 receptor controls development and maturation of late stages of thymocyte subpopulations (2013)
  • Author(s): Tani-Ichi, S., Shimba, A., Wagatsuma, K., Miyachi, H., Kitano, S., Imai, K., Hara, T., and Ikuta, K
  • Journal Title: Proc. Natl. Acad. Sci. U.S.A Volume: 110 Pages: 612-617
  • NAID: 120005241695
• [Journal Article] Posttranscriptional regulation of histone lysine methyltransferase GLP in embryonic male mouse germ cells (2013)
  • Author(s): Deguchi, K., Nagamatsu, G., Miyachi, H., Kato, Y., Morita, S., Kimura, H., Kitano, S., Hatada, I., Saga, Y., Tachibana, M., and Shinkai, Y
  • Journal Title: Biol. Reprod Volume: 88 Pages: 36-36
• [Journal Article] In vivo analysis of Aicda gene regulation : a critical balance between upstream enhancers and intronic silencers governs appropriate expression (2013)
  • Author(s): Huong le, T., Kobayashi, M., Nakata, M., Shioi, G., Miyachi, H., Honjo, T., and Nagaoka, H
  • Journal Title: PLoS One Volume: 8
• [Journal Article] Hypothermia protects against fulminant hepatitis in mice by reducing reactive oxygen species production. (2013)
  • Author(s): Sakurai T, Kudo M, Watanabe T, Itoh K, Higashitsuji H, Arizumi T, Inoue T, Hagiwara S, Ueshima K, Nishida N, Fukumoto M, Fujita J.
  • Journal Title: Dig Dis Volume: 31 Issue: 5-6 Pages: 440-6
  • DOI: 10.1159/000355242
  • Peer Reviewed
• [Journal Article] Epigenetic regulation of sex determination by the histone demethylase Jmjd1a (2013)
  • Author(s): Kuroki S, Matoba S, Akiyoshi M, Matsumura Y, Miyachi H, Mise N, Abe K, Ogura A, Wilhelm D, Koopman P, Nozaki M, Kanai Y, Shinkai Y, and Tachibana, M
  • Journal Title: Science Volume: 341 Issue: 6150 Pages: 1106-1109
  • DOI: 10.1126/science.1239864
  • Peer Reviewed
• [Journal Article] Interleukin-7 receptor controls development and maturation of late stages of thymocyte subpopulations. (2013)
  • Author(s): Tani-Ichi, S., Shimba, A., Wagatsuma, K., Miyachi, H., Kitano, S., Imai, K., Hara, T., and Ikuta, K.
  • Journal Title: Proc. Natl. Acad. Sci. U.S.A. Volume: 110 Issue: 2 Pages: 612-617
  • DOI: 10.1073/pnas.1219242110
  • NAID: 120005241695
  • Peer Reviewed
• [Journal Article] Oscillatory control of factors determining multipotency and fate in mouse neural progenitors (2013)
  • Author(s): I. Imayoshi, A. Isomura, Y. Harima, K. Kawaguchi, H. Kori, H. Miyachi, T. Fujiwara, F. Ishidate, and R. Kageyama
  • Journal Title: Science Volume: 342 Issue: 6163 Pages: 1203-1208
  • DOI: 10.1126/science.1242366
  • Peer Reviewed
• [Journal Article] Overexpression of gankyrin in mouse hepatocytes induces hemangioma by suppressing factor inhibiting hypoxia-inducible factor-1 (FIH-1) and activating hypoxia-inducible factor-1 (2013)
  • Author(s): Y. Liu, H. Higashitsuji, K. Itoh, T. Sakurai, K. Koike, K. Hirota, M. Fukumoto, J. Fujita
  • Journal Title: Biochemical and biophysical research communications Volume: 432 Issue: 1 Pages: 22-27
  • DOI: 10.1016/j.bbrc.2013.01.093
  • Peer Reviewed
• [Journal Article] HTLV-I Tax regulates the cellular proliferation through the down-regulation of PIP3-phosphatase expressions via the NK-kB pathway (2012)
  • Author(s): Fukuda RI, Tsuchiya K, Suzuki K, Itoh K, Fujita J, Utsunomiya A and Tsuji T
  • Journal Title: Int. J. Biochem. Mol. Biol Volume: 3 Pages: 95-104
• [Journal Article] Stromal factors SDF1a, sFRP1, and VEGFD induce dopaminergic neuron differentiation of human pluripotent stem cells (2012)
  • Author(s): Schwartz CM, Tavakoli T, Jamias C, Park C-C, Maudsley S, Martin B, Phillips TM, Yao PJ, Itoh K, Ma W, Rao MS, Arenas E and Mattson MP
  • Journal Title: J. Neurosci. Res Volume: 90 Pages: 1367-1381
• [Journal Article] Cold-inducible RNA-binding protein (Cirp) interacts with Dyrk1b/Mirk and promotes proliferation of immature male germ cells in mice (2012)
  • Author(s): Masuda T, Itoh K, Higashitsuji H, Higashitsuji H, Nakazawa N, Sakurai T, Liu Y, Tokuchi H, Fujita T, Zhao Y, Nishiyama H, Tanaka T, Fukumoto M, Ikawa M, Okabe M and Fujita J
  • Journal Title: Proc. Natl. Acad. Sci. U.S.A Volume: 109 Pages: 10885-10890
• [Journal Article] In situ differentiation of CD8□□ T cells from CD4 T cells in peripheral lymphoid tissues (2012)
  • Author(s): Nambu Y, Hayashi T, Jang KJ, Aoki K, Mano H, Nakano K, Osato M, Takahashi K, Itoh K, Teramukai S, Komori T, Fujita J, Ito Y, Shimizu A and Sugai M
  • Journal Title: Sci. Rep Volume: 2 Pages: 642-642
• [Journal Article] Identification of a novel enhancer that binds Sp1 and contributes to induction of cold-inducible RNA-binding protein (cirp) expression in mammalian cells (2012)
  • Author(s): Sumitomo Y, Higashitsuji H, Higashitsuji H, Liu Y, Fujita T, Sakurai T, Candeias M, Itoh K, Chiba T and Fujita J
  • Journal Title: BMC Biotechnology Volume: 12 Pages: 72-72
• [Journal Article] Identification of IL-7-Producing Cells in Primary and Secondary Lymphoid Organs Using IL-7-GFP Knock-In Mice (2012)
  • Author(s): Hara, T., Shitara, S., Imai, K., Miyachi, H., Kitano, S., Yao, H., Tani-Ichi, S., and Ikuta, K
  • Journal Title: J.Immunol Volume: 189 Pages: 1577-1584
• [Journal Article] In vivo evaluation of PhiC31 recombinase activity in transgenic mice (2012)
  • Author(s): Imayoshi, I., Hirano, K., Kitano, S., Miyachi, H., and Kageyama, R
  • Journal Title: Neurosci.Res Volume: 73 Pages: 106-114
• [Journal Article] A multifunctional teal-fluorescent Rosa26 reporter mouse line for Creand Flp-mediated recombination (2012)
  • Author(s): Imayoshi, I., Hirano, K., Sakamoto, M., Miyoshi, G., Imura, T., Kitano, S., Miyachi, H., and Kageyama, R
  • Journal Title: Neurosci.Res Volume: 73 Pages: 85-91
• [Journal Article] Light-induced silencing of neural activity in Rosa26 knock-in mice conditionally expressing the microbial halorhodopsin eNpHR2.0 (2012)
  • Author(s): Imayoshi, I., Tabuchi, S., Hirano, K., Sakamoto, M., Kitano, S., Miyachi, H., Yamanaka, A., and Kageyama, R
  • Journal Title: Neurosci. Res
• [Journal Article] Role of hepatocyte-derived IL-7 in maintenance of intrahepatic NKT cells and T cells and development of B cells in fetal liver (2012)
  • Author(s): Liang, B., Hara, T., Wagatsuma, K., Zhang, J., Maki, K., Miyachi, H., Kitano, S., Yabe-Nishimura, C., Tani-Ichi, S., and Ikuta, K
  • Journal Title: J.Immunol Volume: 189 Pages: 4444-4450
• [Journal Article] HTLV-I Tax regulates the cellular proliferation through the down-regulation of PIP3-phosphatase expressions via the NK-kB pathway. (2012)
  • Author(s): Fukuda RI, Tsuchiya K, Suzuki K, Itoh K, Fujita J, Utsunomiya A and Tsuji T
  • Journal Title: Int. J. Biochem. Mol. Biol. Volume: 3 Pages: 95-104
  • Peer Reviewed
• [Journal Article] Stromal factors SDF1a, sFRP1, and VEGFD induce dopaminergic neuron differentiation of human pluripotent stem cells. (2012)
  • Author(s): Schwartz CM, Tavakoli T, Jamias C, Park C-C, Maudsley S, Martin B, Phillips TM, Yao PJ, Itoh K, Ma W, Rao MS, Arenas E and Mattson MP
  • Journal Title: J. Neurosci. Res. Volume: 90 Issue: 7 Pages: 1367-1381
  • DOI: 10.1002/jnr.23064
  • Peer Reviewed
• [Journal Article] Cold-inducible RNA-binding protein (Cirp) interacts with Dyrk1b/Mirk and promotes proliferation of immature male germ cells in mice (2012)
  • Author(s): Masuda T, et al.
  • Journal Title: Proc Natl Acad Sci USA Volume: 109 Issue: 27 Pages: 10885-108901
  • DOI: 10.1073/pnas.1121524109
  • Peer Reviewed
• [Journal Article] In situ differentiation of CD8αα T cells from CD4 T cells in peripheral lymphoid tissues (2012)
  • Author(s): Nambu Y., Hayashi T., Jang KJ, Aoki K., Mano H., Nakano K., Osato M., Takabashi K., Itoh K., Teramukai S., Komon T., Fujita J., Ito Y., Shimizu
  • Journal Title: Scientific Reports Volume: 2 Issue: 1 Pages: 930-930
  • DOI: 10.1038/srep00642
  • Peer Reviewed
• [Journal Article] Identification of a novel enhancer that binds Sp1 and contributes to induction of cold-inducible RNA-binding protein (cirp) expression in mammalian cells. (2012)
  • Author(s): Sumitomo Y, Higashitsuji H, Higashitsuji H, Liu Y, Fujita T, Sakurai T, et al
  • Journal Title: BMC Biotechnol Volume: 12 Issue: 1 Pages: 72-72
  • DOI: 10.1186/1472-6750-12-72
  • Peer Reviewed
• [Journal Article] Identification of IL-7-producing cells in primary and secondary lymphoid organs using IL-7-GFP knock-in mice. (2012)
  • Author(s): Hara, T., S. Shitara, K. Imai, H. Miyachi, S. Kitano, H. Yao, S. Tani-Ichi, and K. Ikuta
  • Journal Title: Journal of Immunology Volume: 189 Issue: 4 Pages: 1577-1584
  • DOI: 10.4049/jimmunol.1200586
  • Peer Reviewed
• [Journal Article] In vivo evaluation of PhiC31 recombinase activity in transgenic mice. (2012)
  • Author(s): Imayoshi, I., K. Hirano, S. Kitano, H. Miyachi, and R. Kageyama
  • Journal Title: Neuroscience Research Volume: 73 Issue: 2 Pages: 106-114
  • DOI: 10.1016/j.neures.2012.02.008
  • Peer Reviewed
• [Journal Article] A multifunctional teal-fluorescent Rosa26 reporter mouse line for Cre- and Flp-mediated recombination (2012)
  • Author(s): I.Imayoshi, K.Hirano, M.Sakamoto, G.Miyoshi, T.Imura, S.Kitano, H.Miyachi, R.Kageyama
  • Journal Title: Neuroscience Research Volume: 73 Issue: 1 Pages: 85-91
  • DOI: 10.1016/j.neures.2012.02.003
  • Peer Reviewed
• [Journal Article] Light-induced silencing of neural activity in Rosa26 knock-in mice conditionally expressing the microbial halorhodopsin eNpHR2.0 (2012)
  • Author(s): I.Imayoshi, S.Tabuchi, K.Hirano, M.Sakamoto, S.Kitano, H.Miyachi, A.Yamanaka, R.Kageyama, R.
  • Journal Title: Neuroscience Research Volume: (掲載確定)(印刷中) Issue: 1 Pages: 85-91
  • DOI: 10.1016/j.neures.2012.03.008
  • Peer Reviewed
• [Journal Article] Role of hepatocyte-derived IL-7 in maintenance of intrahepatic NKT cells and T cells and development of B cells in fetal liver. (2012)
  • Author(s): Liang, B., T. Hara, K. Wagatsuma, J. Zhang, K. Maki, H. Miyachi, S. Kitano, C. Yabe-Nishimura, S. Tani-Ichi, and K. Ikuta
  • Journal Title: Journal of Immunology Volume: 189 Issue: 9 Pages: 4444-4450
  • DOI: 10.4049/jimmunol.1201181
  • Peer Reviewed
• [Presentation] CRISPR/Cas9 システムを利用した遺伝子編集マウス作製の実際 (2014)
  • Author(s): 宮地均, 伊藤克彦, 他
  • Organizer: 第61回日本実験動物学会総会