摘要
Cockayne syndrome(CS)is a rare autosomal recessive inherited disorder characterized by a variety of clinical features,including increased sensitivity to sunlight,progressive neurological abnormalities,and the appearance of premature aging.However,the pathogenesis of CS remains unclear due to the limitations of current disease models.Here,we generate integration-free induced pluripotent stem cells(iPSCs)from fibroblasts from a CS patient bearing mutations in CSB/ERCC6 gene and further derive isogenic genecorrected CS-iPSCs(GC-iPSCs)using the CRISPR/Cas9 system.CS-associated phenotypic defects are recapitulated in CS-iPSC-derived mesenchymal stem cells(MSCs)and neural stem cells(NSCs),both of which display increased susceptibility to DNA damage stress.Premature aging defects in CS-MSCs are rescued by the targeted correction of mutant ERCC6.We next map the transcriptomic landscapes in CS-iPSCs and GC-iPSCs and their somatic stem cell derivatives(MSCs and NSCs)in the absence or presence of ultraviolet(UV)and replicative stresses,revealing that defects in DNA repair account for CS pathologies.Moreover,we generate autologous GC-MSCs free of pathogenic mutation under a cGMP(Current Good Manufacturing Practice)-compliant condition,which hold potential for use as improved biomaterials for future stem cell replacement therapy for CS.Collectively,our models demonstrate novel disease features and molecular mechanisms and lay a foundation for the development of novel therapeutic strategies to treat CS.
基金
supported by the National Key Research and Development Program of China(2018YFC2000100)
the Strategic Priority Research Program of the Chinese Academy of Sciences(XDA16010100)
the National Key Research and Development Program of China(2018YFA0107203,2017YFA0103304,2017YFA0102802,2016YFC1000601,2015CB 964800,2014CB910503,and 2018YFA0108500)
the National Natural Science Foundation of China(Grant Nos.81625009,81330008,91749202,91749123,31671429,81671377,81771515,31601109,31601158,81701388,81601233,81822018,81801399,31801010,81801370,81861168034,81571400,and 81771580)
the Program of the Beijing Municipal Science and TechnologyCommission(Z151100003915072)
the Key Research Program of the Chinese Academy of Sciences(KJZDEWTZ-L05)
the Beijing Municipal Commission of Health and Family Planning(PXM2018_026283_000002)
the Advanced Innovation Center for Human Brain Protection(117212,3500-1192012).