Systematic Identification of Barriers to Human iPSC Generation

• Published in: Cell Vol. 158; no. 2; pp. 449 - 461
• Main Authors: Qin, Han, Diaz, Aaron, Blouin, Laure, Lebbink, Robert Jan, Patena, Weronika, Tanbun, Priscilia, LeProust, Emily M., McManus, Michael T., Song, Jun S., Ramalho-Santos, Miguel
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 17.07.2014
• Subjects: ADAM Proteins - metabolism; Cell Adhesion; Cellular Reprogramming; chromatin; dephosphorylation; Embryonic Stem Cells - metabolism; Endocytosis; genes; Humans; induced pluripotent stem cells; Induced Pluripotent Stem Cells - cytology; medicine; RNA interference; somatic cells; transforming growth factor beta; Ubiquitin - metabolism; ubiquitination
• ISSN: 0092-8674; 1097-4172; 1097-4172
• DOI: 10.1016/j.cell.2014.05.040

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. [Display omitted] •Genome-wide RNAi screen reveals known and novel barriers to human cell reprogramming•ADAM29 antagonizes reprogramming via its disintegrin domain•Clathrin-mediated endocytosis antagonizes reprogramming by enhancing TGF-β signaling•Different barrier pathways have a combined effect on reprogramming efficiency A genome-wide screen to identify roadblocks in somatic cell reprogramming coupled with examination of the resulting networks provides a global view on endogenous barriers to making induced pluripotent cells.