title: Systematic identification of barriers to human iPSC generation.
abstract: 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.
authors.affiliation: Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, University of California, San Francisco, San Francisco, CA 94143, USA; Departments of Ob/Gyn and Pathology, Center for Reproductive Sciences, and Diabetes Center, University of California, San Francisco, San Francisco, CA 94143, USA.
authors.forename: Han
authors.initials: H
authors.lastname: Qin
authors.affiliation: Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, University of California, San Francisco, San Francisco, CA 94143, USA; Institute for Human Genetics, University of California, San Francisco, San Francisco, CA 94143, USA.
authors.forename: Aaron
authors.initials: A
authors.lastname: Diaz
authors.affiliation: Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, University of California, San Francisco, San Francisco, CA 94143, USA; Departments of Ob/Gyn and Pathology, Center for Reproductive Sciences, and Diabetes Center, University of California, San Francisco, San Francisco, CA 94143, USA.
authors.forename: Laure
authors.initials: L
authors.lastname: Blouin
authors.affiliation: Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, University of California, San Francisco, San Francisco, CA 94143, USA; Department of Microbiology and Immunology, Diabetes Center, and the WM Keck Center for Noncoding RNAs, University of California, San Francisco, San Francisco, CA 94143, USA.
authors.forename: Robert Jan
authors.initials: RJ
authors.lastname: Lebbink
authors.affiliation: Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, University of California, San Francisco, San Francisco, CA 94143, USA; Department of Microbiology and Immunology, Diabetes Center, and the WM Keck Center for Noncoding RNAs, University of California, San Francisco, San Francisco, CA 94143, USA.
authors.forename: Weronika
authors.initials: W
authors.lastname: Patena
authors.affiliation: Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, University of California, San Francisco, San Francisco, CA 94143, USA; Departments of Ob/Gyn and Pathology, Center for Reproductive Sciences, and Diabetes Center, University of California, San Francisco, San Francisco, CA 94143, USA.
authors.forename: Priscilia
authors.initials: P
authors.lastname: Tanbun
authors.affiliation: Genomics Solution Unit, Agilent Technologies Inc., Santa Clara, CA 95051, USA.
authors.forename: Emily M
authors.initials: EM
authors.lastname: LeProust
authors.affiliation: Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, University of California, San Francisco, San Francisco, CA 94143, USA; Department of Microbiology and Immunology, Diabetes Center, and the WM Keck Center for Noncoding RNAs, University of California, San Francisco, San Francisco, CA 94143, USA.
authors.forename: Michael T
authors.initials: MT
authors.lastname: McManus
authors.affiliation: Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, University of California, San Francisco, San Francisco, CA 94143, USA; Institute for Human Genetics, University of California, San Francisco, San Francisco, CA 94143, USA; Department of Epidemiology and Biostatistics and Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, San Francisco, CA 94143, USA. Electronic address: songj@illinois.edu.
authors.forename: Jun S
authors.initials: JS
authors.lastname: Song
authors.affiliation: Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, University of California, San Francisco, San Francisco, CA 94143, USA; Departments of Ob/Gyn and Pathology, Center for Reproductive Sciences, and Diabetes Center, University of California, San Francisco, San Francisco, CA 94143, USA. Electronic address: miguel.ramalho-santos@ucsf.edu.
authors.forename: Miguel
authors.initials: M
authors.lastname: Ramalho-Santos
mesh_terms: D051722:ADAM Proteins / Q000378:metabolism; D002448:Cell Adhesion; D065150:Cellular Reprogramming; D053595:Embryonic Stem Cells / Q000378:metabolism; D004705:Endocytosis; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D025801:Ubiquitin / Q000378:metabolism
chemical_list: D025801:Ubiquitin; D051722:ADAM Proteins; C122189:ADAM29 protein, human