Zheying Min
Qianzhao Ji
Lingling Geng
Yao Su
Zunpeng Liu
Huifang Hu
Lixia Wang
Weiqi Zhang
Keiichiro Suzuiki
Yu Huang
Puyao Zhang
Tie-Shan Tang
Jing Qu
Yang Yu
Guang-Hui Liu
Jie Qiao
1 Department of Obstetrics and Gynecology, Center for Reproductive Medicine, Peking University Third Hospital, Beijing 100191, China;
2 National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China;
3 State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China;
4 University of Chinese Academy of Sciences, Beijing 100049, China;
5 Advanced Innovation Center for Human Brain Protection, National Clinical Research Center for Geriatric Disorders, Xuanwu Hospital Capital Medical University, Beijing 100053, China;
6 Institute for Stem cell and Regeneration, Chinese Academy of Sciences, Beijing 100101, China;
7 Key Laboratory of Genomic and Precision Medicine, Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing 100101, China;
8 Beijing Institute for Brain Disorders, Beijing 100069, China;
9 Institute for Advanced Co-Creation Studies, Osaka University, Osaka 560-8531, Japan;
10 Graduate School of Engineering Science, Osaka University, Osaka 560-8531, Japan;
11 Department of Medical Genetics, School of Basic Medical Sciences, Peking University Health Science Center, Beijing 100191, China;
12 State Key Laboratory of Membrane Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China;
13 Peking-Tsinghua Center for Life Sciences, Academy for Advanced Interdisciplinary Studies, Peking University, Beijing 100871, China
Funds: The authors acknowledge L. Bai, R. Bai, Q. Chu, J. Lu, S. Ma and Y. Yang for administrative assistance and W. Li, J. Jia and X. Zhang for assistance with animal experiments. This work was 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 Technology Commission (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) and the Advanced Innovation Center for Human Brain Protection (117212, 3500-1192012).
Received Date: 2019-02-19
Rev Recd Date:2019-03-12
Publish Date:2020-01-10
Abstract
Abstract
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.Keywords: Cockayne syndrome,
CRISPR/Cas9,
gene correction,
disease modelling,
mesenchymal stem cell,
neural stem cell
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