TY - JOUR
T1 - Systematic identification of barriers to human Ipsc generation
AU - Qin, Han
AU - Diaz, Aaron
AU - Blouin, Laure
AU - Lebbink, Robert Jan
AU - Patena, Weronika
AU - Tanbun, Priscilia
AU - Leproust, Emily M.
AU - McManus, Michael T.
AU - Song, Jun S.
AU - Ramalho-Santos, Miguel
N1 - We are grateful to Robert Blelloch, Keith Mostov, and Zena Werb for critical reading of the manuscript, Jonathan Weissman, Martin Kampann, and Michael Bassick for discussions and help with library generation, Nikki Shariat for input on the experiment design, Xiaoyin Wang for virus production, Fong Ming Koh for providing the pGAMA vector, Charles Nicolet and Henriette O’Geen for support on NGS, and Tara Rambaldo and Michael Kissner for support with FACS. This work was supported by NWO Rubicon grant 825.06.030 and Veni grant 916.10.138 to R.J.L.; the UCSF Program for Breakthrough Biomedical Research and NIH grants 1U01CA168370 and R01 GM80783 to M.T.M.; NIH R01CA163336 and Sontag Foundation Distinguished Scientist Award to J.S.S.; and CIRM grant RB4-06028 and NIH Director’s New Innovator Award DP2OD4698 to M.R.-S. E.M.L. is an employee of Agilent Technologies.
PY - 2014/7/17
Y1 - 2014/7/17
N2 - Reprogramming of somatic cells to induced pluripotent stem cells (iPSCs) holds enormous promise for regenerative medicine. To elucidate endogenous barriers limiting this process, we systematically dissected human cellular reprogramming by combining a genome-wide RNAi screen, innovative computational methods, extensive single-hit validation, and mechanistic investigation of relevant pathways and networks. We identify reprogramming barriers, including genes involved in transcription, chromatin regulation, ubiquitination, dephosphorylation, vesicular transport, and cell adhesion. Specific a disintegrin and metalloproteinase (ADAM) proteins inhibit reprogramming, and the disintegrin domain of ADAM29 is necessary and sufficient for this function. Clathrin-mediated endocytosis can be targeted with small molecules and opposes reprogramming by positively regulating TGF-β signaling. Genetic interaction studies of endocytosis or ubiquitination reveal that barrier pathways can act in linear, parallel, or feedforward loop architectures to antagonize reprogramming. These results provide a global view of barriers to human cellular reprogramming.
AB - Reprogramming of somatic cells to induced pluripotent stem cells (iPSCs) holds enormous promise for regenerative medicine. To elucidate endogenous barriers limiting this process, we systematically dissected human cellular reprogramming by combining a genome-wide RNAi screen, innovative computational methods, extensive single-hit validation, and mechanistic investigation of relevant pathways and networks. We identify reprogramming barriers, including genes involved in transcription, chromatin regulation, ubiquitination, dephosphorylation, vesicular transport, and cell adhesion. Specific a disintegrin and metalloproteinase (ADAM) proteins inhibit reprogramming, and the disintegrin domain of ADAM29 is necessary and sufficient for this function. Clathrin-mediated endocytosis can be targeted with small molecules and opposes reprogramming by positively regulating TGF-β signaling. Genetic interaction studies of endocytosis or ubiquitination reveal that barrier pathways can act in linear, parallel, or feedforward loop architectures to antagonize reprogramming. These results provide a global view of barriers to human cellular reprogramming.
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U2 - 10.1016/j.cell.2014.05.040
DO - 10.1016/j.cell.2014.05.040
M3 - Article
C2 - 25036638
AN - SCOPUS:84904559751
SN - 0092-8674
VL - 158
SP - 449
EP - 461
JO - Cell
JF - Cell
IS - 2
ER -