Mechanistic analysis of iPSC production using paused iPSC

• Project/Area Number: 26870074
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Developmental biology; General medical chemistry
• Research Institution: University of Tsukuba
• Principal Investigator: Nishimura Ken 筑波大学, 医学医療系, 助教 (80500610)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2015: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2014: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: Klf4 / iPS細胞 / Tcl1 / 発現量依存的

Outline of Final Research Achievements

We succeeded to produce paused iPSCs with different KLF4 expression levels remaining at distinct intermediate stages of reprogramming. Among the genes highly expressing in Klf4-high paused iPSC, Tcl1 related to induction of high pluripotency. The analyses of Klf4-dose dependent transcriptional regulation in paused iPSC indicated that Klf4 binds to Tcl1 promoter dose dependently and lead to histone modifications, resulting in the induction of Tcl1 expression and pluripotency.

Research Products

• [Journal Article] Genetic variability overrides the impact of parental cell-type and determines iPSCs differentiation potential (2016)
  • Author(s): Kyttala A, Moraghebi R, Valensisi C, Kettunen J, Andrus C, Pasumarthy KK, Nakanishi M, Nishimura K, Ohtaka M, Weltner J, Handel BV, Parkkonen O, Sinisalo J, Jalanko A, Hawkins RD, Woods NB, Otonkoski T, Trokovic R
  • Journal Title: Stem Cell Reports Volume: 6 Issue: 2 Pages: 200-212
  • DOI: 10.1016/j.stemcr.2015.12.009
  • NAID: 120007129665
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Functional Neurons Generated from T Cell-Derived Induced Pluripotent Stem Cells (2016)
  • Author(s): Matsumoto T, Fujimori K, Andoh-Noda T, Ando T, Kuzumaki N, Toyoshima M, Tada H, Imaizumi K, Ishikawa M, Yamaguchi R, Isoda M, Zhou Z, Sato S, Kobayashi T, Ohtaka M, Nishimura K, Kurosawa H, Yoshikawa T, Takahashi T, Nakanishi M, Ohyama M, Hattori N, Akamatsu W, Okano H.
  • Journal Title: Stem Cell Reports. Volume: 6 Issue: 3 Pages: 422-35
  • DOI: 10.1016/j.stemcr.2016.01.010
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Klf4遺伝子発現調節によるiPS細胞誘導中間体の作製 (2015)
  • Author(s): 西村健、久武幸司
  • Journal Title: 医学のあゆみ Volume: 255 Pages: 173-174
• [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] 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] An assessment of the effects of ectopic gp91phox expression in XCGD iPSC-derived neutrophils. (2015)
  • Author(s): Lin HT, Masaki H, Yamaguchi T, Wada T, Yachie A, Nishimura K, Ohtaka M, Nakanishi M, Nakauchi H, Otsu M.
  • Journal Title: Mol Ther Methods Clin Dev Volume: 2 Pages: 15046-15046
  • DOI: 10.1038/mtm.2015.46
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Generation of cleidocranial dysplasia-specific human induced pluripotent stem cells in completely serum-, feeder-, and integration-free culture. (2015)
  • Author(s): Yamasaki S, Hamada A, Akagi E, Nakatao H, Ohtaka M, Nishimura K, Nakanishi M, Toratani S, Okamoto T.
  • Journal Title: In Vitro Cell Dev Biol Anim. Volume: 52 Issue: 2 Pages: 252-264
  • DOI: 10.1007/s11626-015-9968-x
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Manipulation of KLF4 expression generates iPSCs paused at successive stages of reprogramming (2014)
  • Author(s): K. Nishimura, T. Kato, C. Chen, L. Oinam, E. Shiomitsu, D. Ayakawa, M. Ohtaka, A. Fukuda, M. Nakanishi, K. Hisatake
  • Journal Title: Stem Cell Reports Volume: 3 Issue: 5 Pages: 915-929
  • DOI: 10.1016/j.stemcr.2014.08.014
  • NAID: 120007136516
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [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 U S A. Volume: 111 Issue: 47 Pages: 16772-7
  • DOI: 10.1073/pnas.1413481111
  • Peer Reviewed / Acknowledgement Compliant
• [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
• [Presentation] Molecular mechanism of KLF4-dose dependent induction of pluripotency during iPSC production (2016)
  • Author(s): K. Nishimura, N.F. Liliani, E. Shiomitsu, M. Ohtaka, A. Fukuda, M. Nakanishi, K. Hisatake
  • Organizer: CiRA/ISSCR International Symposium
  • Place of Presentation: 京都大学百周年時計台記念館
  • Year and Date: 2016-03-22
  • Int'l Joint Research
• [Presentation] Application of novel RNA vector to stem cell engineering (2015)
  • Author(s): Ken Nishimura
  • Organizer: Gene-editing, transgenics & stem cells: Frontiers in medical and molecular therapeutics Workshop
  • Place of Presentation: Dublin, Ireland
  • Year and Date: 2015-09-07
  • Int'l Joint Research / Invited
• [Presentation] Manipulation of KLF4 expression generates iPSCs paused at successive stages of reprogramming (2015)
  • Author(s): K. Nishimura, E. Shiomitsu, N.F. Liliani, T. Kato, C. Chen, M. Ohtaka, A. Fukuda, M. Nakanishi, K. Hisatake
  • 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
  • Organizer: 第14回日本再生医療学会総会
  • Place of Presentation: 横浜
  • Year and Date: 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
  • Organizer: 第14回日本再生医療学会総会
  • Place of Presentation: 横浜
  • Year and Date: 2015-03-19
• [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
  • Organizer: 第14回日本再生医療学会総会
  • Place of Presentation: 横浜
  • Year and Date: 2015-03-19
• [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
  • Organizer: 第37回日本分子生物学会年会
  • Place of Presentation: 横浜
  • Year and Date: 2014-11-25
• [Presentation] Manipulation of Klf4 expression produces paused iPSC stalling at distinct intermediate stages of reprogramming (2014)
  • Author(s): 西村 健
  • Organizer: Keystone Symposia “Stem Cells and Reprogramming”
  • Place of Presentation: Olympic Valley, USA
  • Year and Date: 2014-04-08
• [Remarks] iPS細胞誘導の中間体の作製に成功 ～Klf4遺伝子の発現量によるiPS細胞誘導の調節～
  • URL: https://www.tsukuba.ac.jp/attention-research/p20141003010.html