Abstract
Pluripotent stem cells (PSCs) maintain a low mutation frequency compared with somatic cell types at least in part by preferentially utilizing error-free homologous recombination (HR) for DNA repair. Many endogenous metabolites cause DNA interstrand crosslinks, which are repaired by the Fanconi anemia (FA) pathway using HR. To determine the effect of failed repair of endogenous DNA lesions on PSC biology, we generated iPSCs harboring a conditional FA pathway. Upon FA pathway loss, iPSCs maintained pluripotency but underwent profound G2 arrest and apoptosis, whereas parental fibroblasts grew normally. Mechanistic studies revealed that G2-phase FA-deficient iPSCs possess large γH2AX-RAD51 foci indicative of accrued DNA damage, which correlated with activated DNA-damage signaling through CHK1. CHK1 inhibition specifically rescued the growth of FA-deficient iPSCs for prolonged culture periods, surprisingly without stimulating excessive karyotypic abnormalities. These studies reveal that PSCs possess hyperactive CHK1 signaling that restricts their self-renewal in the absence of error-free DNA repair.
References
Mar 14, 2002·Proceedings of the National Academy of Sciences of the United States of America·Rachel B CervantesPeter J Stambrook
Jan 25, 2005·Nature Cell Biology·Claus Storgaard SørensenThomas Helleday
Jul 23, 2009·Mutation Research·Arleen D Auerbach
Jan 22, 2010·Blood·Asmin TulpuleGeorge Q Daley
May 25, 2010·Stem Cells and Development·Lourdes SerranoJay A Tischfield
Jul 28, 2010·The Journal of Cell Biology·Simona GiuntaStephen P Jackson
Oct 27, 2010·PloS One·Olga MomcilovicGerald Schatten
Dec 25, 2010·The Journal of Clinical Investigation·Raphael CeccaldiJean Soulier
Feb 3, 2011·Molecular Therapy : the Journal of the American Society of Gene Therapy·Eva WarlichAxel Schambach
Feb 11, 2011·Proceedings of the National Academy of Sciences of the United States of America·Kristen L MeerbreyStephen J Elledge
Apr 5, 2011·Nature Methods·Keisuke OkitaShinya Yamanaka
Mar 1, 2012·Blood·Lars U W MüllerDavid A Williams
Jul 4, 2012·Genes & Development·Hyungjin Kim, Alan D D'Andrea
Jan 3, 2013·Stem Cells·Sun K YungMajlinda Lako
Jul 8, 2014·Nature Communications·Guang-Hui LiuJuan Carlos Izpisua Belmonte
Jul 18, 2014·Journal of Virology·Timothy M ChlonSusanne I Wells
May 6, 2015·Stem Cell Reports·Benjamin DannenmannKlaus Schulze-Osthoff
Citations
Jun 23, 2016·Radiation Research·Kalpana MujooTej K Pandita
Sep 23, 2016·Oxidative Medicine and Cellular Longevity·Juan ZhangYuepu Pu
Dec 9, 2016·Expert Review of Hematology·Christen L EbensJohn E Wagner
Sep 24, 2016·Balsaeng'gwa saengsig·Jae-Young LeeKyung-Soon Park
Jul 25, 2017·BioResearch Open Access·Takaharu NegoroAkifumi Matsuyama
Jun 7, 2018·Stem Cells·Haritha VallabhaneniDeborah A Hursh
Aug 18, 2018·Human Gene Therapy·Paula RíoJuan A Bueren
Dec 23, 2016·Clinical Chemistry·Catherine Alix-PanabièresSaid Assou
Feb 23, 2020·Cancers·Ali NowrouziAmir Abdollahi
Oct 31, 2019·Journal of Radiation Research·Mikio ShimadaYoshihisa Matsumoto
Feb 22, 2018·Scientific Reports·Henning HintzscheHelga Stopper
Nov 25, 2020·Cell Stem Cell·Sonya Ruiz-TorresSusanne I Wells
Apr 7, 2019·Biochimica Et Biophysica Acta. Molecular Basis of Disease·Chinnadurai ManiKomaraiah Palle