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

Protein & Cell - Tập 11 Số 1 - Trang 1-22 - 2020
Si Wang1,2,3, Zheying Min2,4, Qianzhao Ji3,5, Lingling Geng1, Yao Su1, Zunpeng Liu6,5, Huifang Hu6,5, Lixia Wang3,5, Weiqi Zhang1,7,8,3,5, Keiichiro Suzuiki9,10, Yu Huang11, Pu-Yao Zhang2, Tie-Shan Tang7,12,5, Jing Qu7,6,5, Yang Yu2, Guang‐Hui Liu1,13,7,3,5, Jie Qiao2,4
1Advanced Innovation Center for Human Brain Protection, National Clinical Research Center for Geriatric Disorders, Xuanwu Hospital Capital Medical University, 100053, Beijing, China
2Department of Obstetrics and Gynecology, Center for Reproductive Medicine, Peking University Third Hospital, 100191, Beijing, China
3National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules Institute of Biophysics, Chinese Academy of Sciences, 100101, Beijing, China
4Peking-Tsinghua Center for Life Sciences, Academy for Advanced Interdisciplinary Studies, Peking University, 100871, Beijing, China
5University of Chinese Academy of Sciences, 100049 Beijing, China
6State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, 100101, Beijing, China
7Institute for Stem Cell and Regeneration, Chinese Academy of Sciences, 100101 Beijing, China
8Key Laboratory of Genomic and Precision Medicine, Beijing Institute of Genomics, Chinese Academy of Sciences, 100101, Beijing, China
9Graduate School of Engineering Science, Osaka University, 560-8531, Osaka, Japan
10Institute for Advanced Co-Creation Studies, Osaka University, 560-8531, Osaka, Japan
11Department of Medical Genetics, School of Basic Medical Sciences, Peking University Health Science Center, 100191, Beijing, China
12State Key Laboratory of Membrane Biology, Institute of Zoology, Chinese Academy of Sciences, 100101 Beijing, China
13Beijing Institute for Brain Disorders, 100069, Beijing, China

Tóm tắt

AbstractCockayne 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 gene-corrected 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.

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