Reprogramming of murine fibroblasts to induced pluripotent stem cells with chemical complementation of Klf4

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 106; no. 22; pp. 8912 - 8917
• Main Authors: Lyssiotis, Costas A, Foreman, Ruth K, Staerk, Judith, Garcia, Michael, Mathur, Divya, Markoulaki, Styliani, Hanna, Jacob, Lairson, Luke L, Charette, Bradley D, Bouchez, Laure C, Bollong, Michael, Kunick, Conrad, Brinker, Achim, Cho, Charles Y, Schultz, Peter G, Jaenisch, Rudolf
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences 02.06.2009; National Acad Sciences
• Subjects: Animals; Benzazepines - pharmacology; Beta cells; Biological Sciences; Cell Differentiation; Cells; Chemicals; Embryonic stem cells; Embryos; Epigenesis, Genetic - drug effects; Fibroblasts; Fibroblasts - cytology; Fibroblasts - drug effects; Gene expression; Genes, Reporter; Homeodomain Proteins - genetics; Indoles - pharmacology; Induced pluripotent stem cells; Kruppel-Like Transcription Factors - genetics; Luciferases - genetics; Mice; Molecules; Nanog Homeobox Protein; Pluripotent stem cells; Pluripotent Stem Cells - cytology; Rodents; Signal transduction; Small Molecule Libraries - pharmacology; Somatic cells; Stem cells
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.0903860106

Ectopic expression of defined transcription factors can reprogram somatic cells to induced pluripotent stem (iPS) cells, but the utility of iPS cells is hampered by the use of viral delivery systems. Small molecules offer an alternative to replace virally transduced transcription factors with chemical signaling cues responsible for reprogramming. In this report we describe a small-molecule screening platform applied to identify compounds that functionally replace the reprogramming factor Klf4. A series of small-molecule scaffolds were identified that activate Nanog expression in mouse fibroblasts transduced with a subset of reprogramming factors lacking Klf4. Application of one such molecule, kenpaullone, in lieu of Klf4 gave rise to iPS cells that are indistinguishable from murine embryonic stem cells. This experimental platform can be used to screen large chemical libraries in search of novel compounds to replace the reprogramming factors that induce pluripotency. Ultimately, such compounds may provide mechanistic insight into the reprogramming process.