Journal Article

Rescue of premature aging defects in Cockayne syndrome stem cells by CRISPR/Cas9-mediated gene correction

Geng, Lingling, Qu, Jing, Tang, Tie-Shan, Hu, Huifang, Wang, Lixia, Huang, Yu, Qiao, Jie, Liu, Zunpeng, Zhang, Weiqi, Yu, Yang, Liu, Guang-Hui, Suzuiki, Keiichiro, Su, Yao, Wang, Si, Ji, Qianzhao, Min, Zheying, Zhang, Puyao

2020

Overview

Cockayne syndrome (CS) is a rare autosomal reces-sive inherited disorder characterized by a variety of clinical features, including increased sensitivity to sun-light, progressive neurological abnormalities, and the appearance of premature aging. However, the pathogenesis of CS remains unclear due to the limita-tions of current disease models. Here, we generate integration-free induced pluripotent stem cells (iPSCs) from broblasts from a CS patient bearing mutations in CSB/ ERCC6 gene and further derive isogenic gene-corrected CS-iPSCs (GC-iPSCs) using the CRISPR/Cas9 system. CS-associated phenotypic defects are recapit-ulated 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)-compli- ant 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 founda- tion for the development of novel therapeutic strategies to treat CS.

• Publisher: Higher Education Press,Oxford University Press (OUP),Springer Nature B.V,Oxford University Press
• Subject: Aging / Aging, Premature / Aging, Premature - pathology / Aging, Premature - therapy / Animal biochemistry / Animals / Autografts / Biochemistry / Biomaterials / Biomedical and Life Sciences / Cell Biology / Cells, Cultured / Cockayne syndrome / Cockayne Syndrome - pathology / Cockayne Syndrome - therapy / Conditioned stimulus / CRISPR / CRISPR-Cas Systems / CRISPR/Cas9 / Cyclic GMP / Cytology / Defects / Deoxyribonucleic acid / Developmental Biology / disease modelling / DNA / DNA damage / DNA Helicases / DNA Helicases - genetics / DNA Repair / DNA Repair Enzymes / DNA Repair Enzymes - genetics / Fibroblasts / gene correction / Gene Editing / Gene Editing - methods / Hereditary diseases / Human Genetics / Humans / Induced Pluripotent Stem Cells / Induced Pluripotent Stem Cells - pathology / Inhibitory postsynaptic potentials / Life Sciences / Male / mesenchymal stem cell / Mesenchymal Stem Cells / Mesenchymal Stem Cells - metabolism / Mesenchymal Stem Cells - pathology / Mesenchyme / Mice / Mice, Inbred NOD / Mice, SCID / Models, Biological / Molecular modelling / Mutation / neural stem cell / Neural Stem Cells / Neural Stem Cells - metabolism / Neural Stem Cells - pathology / Pluripotency / Poly-ADP-Ribose Binding Proteins / Poly-ADP-Ribose Binding Proteins - genetics / Protein Science / QH573-671 / QP501-801 / Research Article / Stem Cells / Targeted Gene Repair / Targeted Gene Repair - methods / Transcriptome / Transcriptomics