Analysis of the transcription factor network during the early stage of iPSC production

• Project/Area Number: 25460354
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: General medical chemistry
• Research Institution: University of Tsukuba
• Principal Investigator: Hisatake Koji 筑波大学, 医学医療系, 教授 (70271236)
• Co-Investigator(Kenkyū-buntansha): NISHIMURA KEN 筑波大学, 医学医療系, 助教 (80500610)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2014: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2013: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: 転写因子 / 遺伝子発現制御 / リプログラミング / iPS細胞 / 間葉上皮転換 / 遺伝子発現 / クロマチン / iPS 細胞 / エピジェネティクス / 細胞初期化 / センダイウイルス

Outline of Final Research Achievements

Genome-wide expression analyses were used to identify transcription factors whose expression decrease at the early stage or reprogramming. They were further narrowed down by other data base to select for the genes that are likely to be regulated directly by Oct4 and Klf4. The selection resulted in 38 candidate genes whose expression changes are potentially important for the early stage of reprogramming. siRNA-mediated knockdown and over-expression of these genes showed that down-regulation of Osr1、Ebf1、Ebf3、Meox2、Smardc3 and Prrx is essential at the early stage of reprogramming. The involvement of these genes in EMT or MET was examined by the used of NMuMG cells, and the Ebf3 was found to have an activity in promoting EMT.

Research Products

• [Journal Article] The RNA-binding Complexes, NF45-NF90 and NF45-NF110, Associate Dynamically with the c-fos Gene and Function as Transcriptional Coactivators (2015)
  • Author(s): Nakadai T, Fukuda A, Shimada M, Nishimura K, Hisatake K
  • Journal Title: The Journal of Biological Chemistry Volume: 290 Issue: 44 Pages: 26832-26845
  • DOI: 10.1074/jbc.m115.688317
  • NAID: 120007135696
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] A Safeguard System for Induced Pluripotent Stem Cell-Derived Rejuvenated T Cell Therapy. (2015)
  • Author(s): Ando M, Nishimura T, Yamazaki S, Yamaguchi T, Kawana-Tachikawa A, Hayama T, Nakauchi Y, Ando J, Ota Y, Takahashi S, Nishimura K, Ohtaka M, Nakanishi M, Miles JJ, Burrows SR, Brenner MK, Nakauchi H.
  • Journal Title: Stem Cell Reports. Volume: 5(4) Issue: 4 Pages: 597-608
  • DOI: 10.1016/j.stemcr.2015.07.011
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Association of the Winged Helix Motif of the TFIIEα Subunit of TFIIE with Either the TFIIEβ Subunit or TFIIB Distinguishes Its Functions in Transcription. (2015)
  • Author(s): Tanaka, A., Akimoto, Y., Kobayashi, S., Hisatake, K., Hanaoka, F., and Ohkuma, Y.
  • Journal Title: Genes Cells Volume: 20 Issue: 3 Pages: 203-216
  • DOI: 10.1111/gtc.12212
  • Peer Reviewed / Open Access
• [Journal Article] Interspecific in vitro assay for the chimera-forming ability of human pluripotent stem cells. (2015)
  • Author(s): Masaki H, Kato-Itoh M, Umino A, Sato H, Hamanaka S, Kobayashi T, Yamaguchi T, Nishimura K, Ohtaka M, Nakanishi M, Nakauchi H.
  • Journal Title: Development. Volume: 142(18) Pages: 3222-30
  • DOI: 10.1242/dev.124016
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Prediction of inter-individual differences in hepatic functions and drug sensitivity by using human iPS-derived hepatocytes (2014)
  • Author(s): Takayama K, Morisaki Y, Kuno S, Nagamoto Y, Harada K, Furukawa N, Ohtaka M, Nishimura K, Imagawa K, Sakurai F, Tachibana M, Sumazaki R, Noguchi E, Nakanishi M, Hirata K, Kawabata K, Mizuguchi H
  • Journal Title: Proc. Natl. Acad. Sci. USA Volume: 111 Pages: 16772-16777
  • Peer Reviewed
• [Journal Article] Advanced Feeder-Free Generation of iPSC Directly From Blood Cells (2014)
  • Author(s): Trokovic R, Weltner J, Nishimura K, Ohtaka M, Nakanishi M, Salomaa V, Jalanko A, Otonkoski T, A Kyttälä
  • Journal Title: Stem Cells Trans. Med. Volume: 3 Issue: 12 Pages: 1402-1409
  • DOI: 10.5966/sctm.2014-0113
  • Peer Reviewed
• [Journal Article] Manipulation of KLF4 Expression Generates iPSCs Paused at Successive Stages of Reprogramming (2014)
  • Author(s): Nishimura K, Kato T, Chen C, Oinam L, Shiomitsu E, Ayakawa D, Ohtaka M, Fukuda A, Nakanishi M, Hisatake K
  • Journal Title: Stem Cell Reports Volume: 3 Pages: 915-929
  • NAID: 120007136516
  • Peer Reviewed / Open Access
• [Journal Article] Heterogeneous Nuclear Ribonucleoprotein R Cooperates with Mediator to Facilitate Transcription Reinitiation on the c-Fos Gene (2013)
  • Author(s): A. Fukuda, M. Shimada, T. Nakadai, K. Nishimura, K. Hisatake
  • Journal Title: PLOS ONE Volume: Vol. 8 Issue: 8 Pages: e72496-e72496
  • DOI: 10.1371/journal.pone.0072496
  • NAID: 120007136844
  • Peer Reviewed
• [Journal Article] The Paired-box Domain Transcription Factor Pax6 Binds to the Upstream Region of the TRAP Gene Promoter and Suppresses RANKL-induced Osteoclast Differentiation (2013)
  • Author(s): Kogawa M, Hisatake K, Atkins GJ, Findlay DM, Enoki Y, Sato T, Gray PC, Kanesaki-Yatsuka Y, Anerson PH, Wada S, Kato N, Fukuda A, Katayama S, Tsujimoto M, Yoda T, Suda T, Okazaki Y, Matsumoto M
  • Journal Title: J Biol Chem Volume: 288 Issue: 43 Pages: 31299-31312
  • DOI: 10.1074/jbc.m113.461848
  • Peer Reviewed
• [Presentation] Molecular mechanism of KLF4-dose dependent induction of pluripotency during iPSC production (2016)
  • Author(s): Nishimura K, Liliani NF, Shiomitsu E, Ohtaka M, Fukuda A, Nakanishi M, Hisatake K
  • Organizer: CiRA/ISSCR International Symposium
  • Place of Presentation: 京都大学 iPS 細胞研究所 (CiRA)
  • Year and Date: 2016-03-22
  • Int'l Joint Research
• [Presentation] Manipulation of KLF4 expression reveals a mechanistic insight into an intermediate state of reprogramming (2015)
  • Author(s): Hisatake K, Lilliani NF, Shiomitsu E, Ohtaka M, Fukuda A, Nakanishi M, Nishimura K
  • Organizer: 第38回分子生物学会
  • Place of Presentation: 神戸国際会議場
  • Year and Date: 2015-12-01
  • Invited
• [Presentation] Manipulation of KLF4 expression generates iPSCs paused at successive stages of reprogramming (2015)
  • Author(s): Nishimura K, Shiomitsu E, Liliani N.F., Kato T, Chen C, Ohtaka M, Fukuda A, Nakanishi M, Hisatake K
  • Organizer: International Society of Stem Cell Research 13th Annual Meeting
  • Place of Presentation: Stockholm, Sweden
  • Year and Date: 2015-06-24
  • Int'l Joint Research
• [Presentation] Paused iPSCを用いたiPS細胞誘導過程の解析 (2015)
  • Author(s): 西村健、加藤哲男、陳晨、Lalhaba Oinam、塩満鋭美、大高真奈美、福田綾、中西真人、久武幸司
  • Organizer: 第14回日本再生医療学会
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2015-03-19 – 2015-03-21
• [Presentation] iPS細胞誘導初期に関与する転写因子の同定とその機能解析 (2015)
  • Author(s): 加藤哲男、西村健、Nguyen Thuy Linh、大高真奈美、福田綾、中西真人、久武幸司
  • Organizer: 第14回日本再生医療学会
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2015-03-19 – 2015-03-21
• [Presentation] 細胞の初期化におけるリプログラム因子の機能解析 (2015)
  • Author(s): 塩満鋭美、西村健、Lalhaba Oinam、Fransiska Liliani、福田綾、久武幸司
  • Organizer: 第14回日本再生医療学会
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2015-03-19 – 2015-03-21
• [Presentation] Klf4遺伝子発現量調節による多能性が異なるiPS細胞の誘導 (2014)
  • Author(s): 西村健、加藤哲男、C. Chen、L. Oinam、塩満鋭美、大高真奈美、福田綾、中西真人、久武幸司
  • Organizer: 第37回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2014-11-25 – 2014-11-27
• [Presentation] 初期化が途中で停止した細胞を用いたiPS細胞誘導過程の解析 (2014)
  • Author(s): 西村健、加藤哲男、Lalhaba Oinam、塩満鋭美、成田孝仁、大高真奈美、福田綾、中西真人、久武幸司
  • Organizer: 第13回日本再生医療学会総会
  • Place of Presentation: 京都、国立京都国際会館
• [Presentation] iPS細胞発現誘導初期における体細胞特異的遺伝子の発現減少と誘導効率への影響 (2014)
  • Author(s): 加藤哲男、Lalhaba Oinam、塩満鋭美、福田綾、中西真人、西村健、久武幸司
  • Organizer: 第13回日本再生医療学会総会
  • Place of Presentation: 京都、国立京都国際会館
• [Presentation] Taking snap shots during iPSC production by SeVdp vectors (2013)
  • Author(s): Nishimura K, Kato T, Oinam L, Narita T, Ohtaka M, Fukuda A, Nakanishi M, Hisatake K.
  • Organizer: International Society of Stem Cell Research 11th Annual Meeting
  • Place of Presentation: 米国、ボストン、Boston Convention & Exhibition Center
• [Presentation] Regulated Induction of Partially Reprogrammed iPS Cells by SeVdp Vectors (2013)
  • Author(s): Hisatake K, Kato T, Oinam L, Narita T, Ohtaka M, Fukuda A, Nakanishi M, Nishimura K.
  • Organizer: Cold Spring Harbor Laboratory meeting on Stem Cell Biology
  • Place of Presentation: 米国、ニューヨーク、Cold Spring Harbor Laboratory
• [Presentation] Manipulation of Klf4 expression produces snap shots of iPSC induction (2013)
  • Author(s): Nishimura K, Kato T, Oinam L, Ohtaka M, Fukuda A, Nakanishi M, Hisatake K.
  • Organizer: CSHAsia 2013 Conference on CSHA/ISSCR Joint Meeting on Stem Cells in Science and Medicine
  • Place of Presentation: 中国、江蘇省、江蘇国際会議センター
• [Presentation] Klf4遺伝子発現量による初期化が途中で停止した細胞の誘導 (2013)
  • Author(s): 西村健、加藤哲男、Oinam Lalhaba、塩満鋭美、成田孝仁、大高真奈美、福田綾、中西真人、久武幸司
  • Organizer: 第36回日本分子生物学会年会
  • Place of Presentation: 神戸、神戸ポートアイランド
• [Presentation] iPS細胞誘導初期におけるメカニズム解析 (2013)
  • Author(s): 加藤哲男、西村健、塩満鋭美、綾川大介、福田綾、久武幸司
  • Organizer: 第36回日本分子生物学会年会
  • Place of Presentation: 神戸、神戸ポートアイランド
• [Presentation] Analysis of the Partially Reprogrammed iPS Cells (2013)
  • Author(s): Shiomitsu E, Nishimura K, Kato T, Oinam L, Fukuda A, Hisatake K
  • Organizer: The 12th NTU-Japan International Student Mini-Symposium on Life Science
  • Place of Presentation: 台湾、台北、国立台湾大学
• [Remarks] 遺伝子制御学研究室
  • URL: http://www.md.tsukuba.ac.jp/basic-med/biochem/gene/index.html
• [Remarks] 筑波大学 注目の研究
  • URL: https://www.tsukuba.ac.jp/attention-research/p20141003010.html