TY - JOUR
T1 - A molecular roadmap of reprogramming somatic cells into iPS cells
AU - Polo, Jose M.
AU - Anderssen, Endre
AU - Walsh, Ryan M.
AU - Schwarz, Benjamin A.
AU - Nefzger, Christian M.
AU - Lim, Sue Mei
AU - Borkent, Marti
AU - Apostolou, Effie
AU - Alaei, Sara
AU - Cloutier, Jennifer
AU - Bar-Nur, Ori
AU - Cheloufi, Sihem
AU - Stadtfeld, Matthias
AU - Figueroa, Maria Eugenia
AU - Robinton, Daisy
AU - Natesan, Sridaran
AU - Melnick, Ari
AU - Zhu, Jinfang
AU - Ramaswamy, Sridhar
AU - Hochedlinger, Konrad
N1 - Funding Information:
We thank members of the Hochedlinger and Ramaswamy labs for helpful suggestions and critical reading of the manuscript. We thank Laura Prickett and Kat Folz-Donahue at the MGH/HSCI flow cytometry core and Flowcore Monash for expert cell sorting; Paul Lacaze, Danielle Evans, and Michelle Garred at Millenium Science for support with the Fluidigm experiments; and Lucy Dagostino from Lifetech Australia for help in miRNA assays. We are grateful to Ben Wittner for help with formatting and uploading genome-wide data. Support to J.M.P. was from an ECOR postdoctoral fellowship, Monash Larkins Program, and a NHMRC CDF. J.Z. was supported by the Division of Intramural Research, National Institute of Allergy and Infectious Diseases, NIH, USA. K.H. was supported by the NIH (DP2OD003266 and R01HD058013). S.R. was supported by an HHMI Physican-Scientist Early Career Award.
PY - 2012/12/21
Y1 - 2012/12/21
N2 - Factor-induced reprogramming of somatic cells into induced pluripotent stem cells (iPSCs) is inefficient, complicating mechanistic studies. Here, we examined defined intermediate cell populations poised to becoming iPSCs by genome-wide analyses. We show that induced pluripotency elicits two transcriptional waves, which are driven by c-Myc/Klf4 (first wave) and Oct4/Sox2/Klf4 (second wave). Cells that become refractory to reprogramming activate the first but fail to initiate the second transcriptional wave and can be rescued by elevated expression of all four factors. The establishment of bivalent domains occurs gradually after the first wave, whereas changes in DNA methylation take place after the second wave when cells acquire stable pluripotency. This integrative analysis allowed us to identify genes that act as roadblocks during reprogramming and surface markers that further enrich for cells prone to forming iPSCs. Collectively, our data offer new mechanistic insights into the nature and sequence of molecular events inherent to cellular reprogramming.
AB - Factor-induced reprogramming of somatic cells into induced pluripotent stem cells (iPSCs) is inefficient, complicating mechanistic studies. Here, we examined defined intermediate cell populations poised to becoming iPSCs by genome-wide analyses. We show that induced pluripotency elicits two transcriptional waves, which are driven by c-Myc/Klf4 (first wave) and Oct4/Sox2/Klf4 (second wave). Cells that become refractory to reprogramming activate the first but fail to initiate the second transcriptional wave and can be rescued by elevated expression of all four factors. The establishment of bivalent domains occurs gradually after the first wave, whereas changes in DNA methylation take place after the second wave when cells acquire stable pluripotency. This integrative analysis allowed us to identify genes that act as roadblocks during reprogramming and surface markers that further enrich for cells prone to forming iPSCs. Collectively, our data offer new mechanistic insights into the nature and sequence of molecular events inherent to cellular reprogramming.
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U2 - 10.1016/j.cell.2012.11.039
DO - 10.1016/j.cell.2012.11.039
M3 - Article
C2 - 23260147
AN - SCOPUS:84871586080
VL - 151
SP - 1617
EP - 1632
JO - Cell
JF - Cell
SN - 0092-8674
IS - 7
ER -