Rescue of premature aging defects in Cockayne Syndrome Derived Stem Cells by Targeted Gene Correction

Cockayne Syndrome (CS) is a rare autosomal recessive inherited disorder characterized by a variety of clinical features including sensitivity to sunlight, progressive neurological abnormalities and appearance of premature aging. However, the pathogenesis of CS yet remains unclear due to the limitations of current disease models. Here we generate integration free-induced pluripotent stem cells (iPSCs) from CS patient fibroblast bearing mutations in CSB/ERCC6 and further derive isogenic gene corrected (GC)-iPSCs using CRISPR/Cas9 system. CS cellular phenotypes are recapitulated in CS mesenchymal stem cells (MSCs) and neural stem cells (NSCs), both of which display compromised susceptibility to DNA damage stress. We next map the transcriptome landscapes of CS- and GC-iPSCs and their adult stem/progenitor cell derivatives under ultraviolet (UV) and replicative stress, indicating the defects in DNA repair in confronting the acute and chronic stress contribute to CS-disease pathology. Through RNA sequencing analysis, we also identify a panel of potential targets for treating CS. Moreover, we generate clinical GC-MSCs displaying safer and superior stem cell activity in vitro and in vivo, which may provide better cell materials for future stem cell replacement therapy in CS-disease. Our models serve to facilitate the discovery of novel disease features and molecular mechanism, as well as lay a foundation for development of novel therapeutic strategies to treat CS.