title: Deficient DNA damage response and cell cycle checkpoints lead to accumulation of point mutations in human embryonic stem cells.
abstract: Human embryonic stem cells (hESCs) tend to lose genomic integrity during long periods of culture in vitro and to acquire a cancer-like phenotype. In this study, we aim at understanding the contribution of point mutations to the adaptation process and at providing a mechanistic explanation for their accumulation. We observed that, due to the absence of p21/Waf1/Cip1, cultured hESCs lack proper cell cycle checkpoints and are vulnerable to the kind of DNA damage usually repaired by the highly versatile nucleotide excision repair (NER) pathway. In response to UV-induced DNA damage, the majority of hESCs succumb to apoptosis; however, a subpopulation continues to proliferate, carrying damaged DNA and accumulating point mutations with a typical UV-induced signature. The UV-resistant cells retain their proliferative capacity and potential for pluripotent differentiation and are markedly less apoptotic to subsequent UV exposure. These findings demonstrate that, due to deficient DNA damage response, the modest NER activity in hESCs is insufficient to prevent increased mutagenesis. This provides for the appearance of genetically aberrant hESCs, paving the way for further major genetic changes.
authors: Institute for Cancer Studies, University of Sheffield, Sheffield, United Kingdom.
authors: Nevila
authors: N
authors: Hyka-Nouspikel
authors: Joëlle
authors: J
authors: Desmarais
authors: Paul J
authors: PJ
authors: Gokhale
authors: Mark
authors: M
authors: Jones
authors: Mark
authors: M
authors: Meuth
authors: Peter W
authors: PW
authors: Andrews
authors: Thierry
authors: T
authors: Nouspikel
mesh_terms: D017209:Apoptosis / Q000235:genetics; D059447:Cell Cycle Checkpoints / Q000235:genetics*; D048708:Cell Growth Processes / Q000235:genetics; D002478:Cells, Cultured; D004249:DNA Damage; D004260:DNA Repair; D053595:Embryonic Stem Cells / Q000166:cytology / Q000502:physiology*; D006801:Humans; D017354:Point Mutation