幹細胞、再生医療

未分類

2024.03.03

iPS cells
再生医療

iPS cells

iPS was first made from skin cells. Any cells can be used but now routinely iPS cells can be produced from blood samples (lymphocytes are used).

A New Era of Medicine with iPS Cells – Lecture by Professor Shinya Yamanaka NobelPrizeII チャンネル登録者数 1.5万人

4 genes Yamanaka used for his paper: Oct3/4, Sox2, Klf4, c-Myc (later he found that c-Myc was not needed)

A New Era of Medicine with iPS Cells – Lecture by Professor Shinya Yamanaka (1:17:29) NobelPrizeII チャンネル登録者数 1.5万人
Induction of Pluripotency by Defined Factors (1:09:50) NIH VideoCast チャンネル登録者数 3.92万人 NIH Director’s Wednesday Afternoon Lectures Air date: Thursday, January 14, 2010, 3:00:00 PM

Review articles

Review Article GLIS1-3: emerging roles in reprogramming, stem and progenitor cell differentiation and maintenance David W. Scoville, Hong Soon Kang, Anton M. Jetten Published: 27 September 2017. doi: 10.21037/sci.2017.09.01 The GLI Similar 1-3 (GLIS) proteins form a subfamily of Krüppel-like zinc finger transcription factors that are closely-related to the GLI and ZIC subfamilies (1–8). Members of these three subfamilies share a highly homologous DNA binding domain (DBD) consisting of five Cys₂His₂-type zinc finger motifs. However, these proteins exhibit little homology outside their DBD region. … Initial overexpression of OCT3/4 (POU5F1), SOX2, and KLF4 (OSK) are widely used for the reprogramming of somatic cells into iPSCs (32). However, the efficiency of generating iPSCs is very low, which has been attributed to difficulties in overcoming epigenetics barriers in the starting cell (33). Co-expression of C-MYC increases the efficiency, but also enhances the potential tumorigenicity of iPSC-derived differentiated cells. Recently, using a screen analyzing 1,437 transcription factors for their ability to promote reprogramming efficiency, GLIS1 was found to greatly enhance the number of iPSC colonies generated when co-expressed with OSK (referred to as OSKG) in either human or mouse dermal fibroblasts (29,34). Inversely, down-regulation of GLIS1 expression by shRNAs reduced the OSK-induced generation of iPSC colonies in mouse fibroblasts suggesting that endogenous GLIS1 is able to promote OSK-mediated reprogramming.
Ye L, Swingen C, Zhang J. Induced pluripotent stem cells and their potential for basic and clinical sciences. Curr Cardiol Rev. 2013 Feb 1;9(1):63-72. doi: 10.2174/157340313805076278. PMID: 22935022; PMCID: PMC3584308. Later, it was shown that iPS cells can be generated from fibroblasts by viral integration of Oct4/Sox2/Klf4 without c-Myc [4]. Although these iPS cells showed reduced tumorigenicity in chimeras and progeny mice, the reprogramming process is much slower, and efficiency is substantially reduced.
Schmidt R, Plath K. The roles of the reprogramming factors Oct4, Sox2 and Klf4 in resetting the somatic cell epigenome during induced pluripotent stem cell generation. Genome Biol. 2012 Oct 22;13(10):251. doi: 10.1186/gb-2012-13-10-251. PMCID: PMC3491406. The most well known of these enhancer factors is c-Myc, which was added alongside O, S and K in the original reprogramming experiment but later shown to be dispensible [1,5,9,10,15,16].
Takahashi K, Mitsui K, Yamanaka S. Role of ERas in promoting tumour-like properties in mouse embryonic stem cells. Nature. 2003 May 29;423(6939):541-5. doi: 10.1038/nature01646. PMID: 12774123. (Yamanaka moved to Kyoto University in 2005. Yamanaka asked Kazu Takahashi to take over the project (24 candidate genes for reprogramming). Yamanaka knew that the project was risky but thought it was ok for the next a couple of years without papers as Takahashi published this Nature paper.(https://www.youtube.com/watch?v=AD1sZU1yk-Y 33:07))

Original articles

Direct reprogramming of somatic cells is promoted by maternal transcription factor Glis1 Momoko Maekawa, Kei Yamaguchi, Tomonori Nakamura, Ran Shibukawa, Ikumi Kodanaka, Tomoko Ichisaka, Yoshifumi Kawamura, Hiromi Mochizuki, Naoki Goshima, and Shinya Yamanaka Nature, Volume: 474, Pages: 225-229, Date published: 09 June 2011, DOI: 10.1038/nature10106
1. 転写因子Ｇｌｉｓ１により安全なｉＰＳ細胞の高効率作製に成功平成２３年６月９日京都大学ｉＰＳ細胞研究所（ＣｉＲＡ） 転写因子Ｇｌｉｓ１の導入によりマウス／ヒトｉＰＳ細胞の樹立効率が大幅に改善される。Ｇｌｉｓ１は、不完全に初期化された細胞の増殖を抑制し、完全に初期化されたｉＰＳ細胞のみを増殖促進させる。
Nakagawa M, Koyanagi M, Tanabe K, Takahashi K, Ichisaka T, Aoi T, Okita K, Mochiduki Y, Takizawa N, Yamanaka S. Generation of induced pluripotent stem cells without Myc from mouse and human fibroblasts. Nat Biotechnol. 2008;26:101–106. doi: 10.1038/nbt1374.
Wernig M, Meissner A, Cassady JP, Jaenisch R. c-Myc is dispensable for direct reprogramming of mouse fibroblasts. Cell Stem Cell. 2008;2:10–12. doi: 10.1016/j.stem.2007.12.001.

Other papers

Tapia, N., MacCarthy, C., Esch, D. et al. Dissecting the role of distinct OCT4-SOX2 heterodimer configurations in pluripotency. Sci Rep 5, 13533 (2015). https://doi.org/10.1038/srep13533