Rescue of premature aging defects in Cockayne syndrome stem cells by CRISPR/Cas9-mediated gene correction
Rescue of premature aging defects in Cockayne syndrome stem cells by CRISPR/Cas9-mediated gene correction
复制标题
通过 CRISPR/Cas9 介导的基因校正来挽救科凯恩综合征干细胞的早衰缺陷。
DOI：
10.1007/s13238-019-0623-2
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发表时间：
2020-01-01
期刊：
影响因子：
21.1
通讯作者：
Qiao, Jie
中科院分区：
文献类型：
--
作者：
Wang, Si;Min, Zheying;Qiao, Jie
Cockayne综合征（CS）是一种罕见的常染色体隐性遗传性疾病，其特征在于多种临床特征，包括对阳光的敏感性增加、进行性神经异常和过早衰老的外观。然而，由于当前疾病模型的局限性，CS的发病机制仍不清楚。在这里，我们从来自携带CSB/ERCC 6基因突变的CS患者的成纤维细胞产生无整合的诱导多能干细胞（iPSC），并使用CRISPR/Cas9系统进一步衍生等基因基因校正的CS-iPSC（GC-iPSC）。CS相关的表型缺陷在CS-iPSC衍生的间充质干细胞（MSC）和神经干细胞（NSC）中重现，这两者都显示出对DNA损伤应激的易感性增加。CS-MSC中的过早衰老缺陷通过突变ERCC 6的靶向校正而得以挽救。接下来，我们绘制了CS-iPSC和GC-iPSC及其体干细胞衍生物（MSC和NSC）在不存在或存在紫外线（UV）和复制应激的情况下的转录组学景观，揭示了DNA修复缺陷导致CS病理。此外，我们产生自体GC-MSC的致病性突变的cGMP（现行良好的生产规范）符合条件下，保持作为未来的干细胞替代治疗CS的改进的生物材料的潜力。总的来说，我们的模型展示了新的疾病特征和分子机制，并为开发治疗CS的新治疗策略奠定了基础。
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 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.