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. Transcriptome analysis of human CS- and gene corrected (GC)-iPSCs, and CS-,GC-MSCs upon UV treatment or under replicative senescence stress, as well as CS-, GC-NSCs upon UV treatment.

科凯恩综合征（Cockayne Syndrome, CS）是一种罕见的常染色体隐性遗传性疾病，以日光敏感性、进行性神经异常及早衰表现等多种临床特征为典型表征。然而，受限于当前已有的疾病模型，科凯恩综合征的发病机制至今仍未明确。本研究从携带CSB/ERCC6突变的科凯恩综合征患者成纤维细胞中构建无整合型诱导多能干细胞（integration free-induced pluripotent stem cells, iPSCs），并通过CRISPR/Cas9系统进一步获得同基因校正（GC）-iPSCs。科凯恩综合征的细胞表型可在CS间充质干细胞（mesenchymal stem cells, MSCs）与神经干细胞（neural stem cells, NSCs）中重现，两类细胞均表现出DNA损伤应激易感性受损的特征。随后，我们绘制了紫外线（ultraviolet, UV）及复制应激下，CS与GC-iPSCs及其成体干/祖细胞衍生物的转录组图谱，结果表明，应对急性与慢性应激时DNA修复缺陷是介导科凯恩综合征病理发生的核心因素。通过RNA测序分析，本研究还筛选出一组可用于治疗科凯恩综合征的潜在靶点。此外，本研究构建了在体内外均表现出更高安全性与更优干细胞活性的临床级GC-MSCs，可为未来科凯恩综合征的干细胞替代治疗提供更为优质的细胞材料。本研究所构建的模型有助于发掘新型疾病特征与分子机制，同时为开发科凯恩综合征的新型治疗策略奠定了坚实基础。本数据集涵盖紫外线处理或复制衰老应激下的人源CS及基因校正（GC）iPSCs、CS与GC-MSCs，以及紫外线处理后的CS与GC-NSCs的转录组分析数据。