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A Small-Molecule Inhibitor of Tgf-β Signaling Replaces Sox2 in Reprogramming by Inducing Nanog
Justin K. Ichida1, 3, 6, Joel Blanchard1, 6, Kelvin Lam1, 6, Esther Y. Son1, 2, 3, 5, 6, Julia E. Chung1, 2, 3, Dieter Egli1, 3, Kyle M. Loh1, Ava C. Carter1, 3, Francesco P. Di Giorgio1, 3, Kathryn Koszka1, 3, Danwei Huangfu1, Hidenori Akutsu4, David R. Liu5, Lee L. Rubin1, , and Kevin Eggan1, 2, 3, ,
1 Harvard Stem Cell Institute, Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA 02138, USA
2 Department of Molecular and Cellular Biology, Harvard University, Cambridge, MA 02138, USA
3 Stowers Medical Institute, 1000 East 50th Street. Kansas City, MO 64110, USA
4 Department of Reproductive Biology, National Research Institute for Child Health and Development, 2-10-1 Okura, Setagaya, Tokyo 157-8535
5 Howard Hughes Medical Institute and the Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA 02138, USA
The combined activity of three transcription factors can reprogram adult cells into induced pluripotent stem cells (iPSCs). However, the transgenic methods used for delivering reprogramming factors have raised concerns regarding the future utility of the resulting stem cells. These uncertainties could be overcome if each transgenic factor were replaced with a small molecule that either directly activated its expression from the somatic genome or in some way compensated for its activity. To this end, we have used high-content chemical screening to identify small molecules that can replace Sox2 in reprogramming. We show that one of these molecules functions in reprogramming by inhibiting Tgf-β signaling in a stable and trapped intermediate cell type that forms during the process. We find that this inhibition promotes the completion of reprogramming through induction of the transcription factor Nanog.
