Whole-genome mutational burden analysis of three pluripotency induction methods.
Whole-genome mutational burden analysis of three pluripotency induction methods.
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DOI：
10.1038/ncomms10536
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发表时间：
2016-02-19
影响因子：
16.6
通讯作者：
Loring JF
中科院分区：
文献类型：
--
作者：
Bhutani K;Nazor KL;Williams R;Tran H;Dai H;Džakula Ž;Cho EH;Pang AWC;Rao M;Cao H;Schork NJ;Loring JF
人们担心诱导多能性的应激可能导致诱导多能干细胞（iPSC）系中的有害DNA突变，这将损害它们用于细胞疗法的用途。在这里，我们报告比较基因组分析的九个同基因iPSC线使用三种重编程方法：整合逆转录病毒载体，非整合仙台病毒和合成的mRNA。我们使用全基因组测序和从头基因组定位来鉴定单核苷酸变异、插入和缺失以及结构变异。我们的研究结果显示，iPSC中存在中等数量的变体，这些变体在亲本成纤维细胞中并不明显，这可能是重编程所致。不同的重编程方法在变异的总数和类型上只有很小的差异。最重要的是，彻底的基因组分析表明，这些变异通常是良性的。我们的结论是，重编程过程不太可能引入使细胞不适合治疗的变异。 人们担心重编程可能会引入DNA突变。Bhutani等人采用了三种不同的重编程方法，并使用比较性全基因组分析确定了重编程细胞与原始成纤维细胞不同的核苷酸变异，但没有一种具有致癌潜力。
There is concern that the stresses of inducing pluripotency may lead to deleterious DNA mutations in induced pluripotent stem cell (iPSC) lines, which would compromise their use for cell therapies. Here we report comparative genomic analysis of nine isogenic iPSC lines generated using three reprogramming methods: integrating retroviral vectors, non-integrating Sendai virus and synthetic mRNAs. We used whole-genome sequencing and de novo genome mapping to identify single-nucleotide variants, insertions and deletions, and structural variants. Our results show a moderate number of variants in the iPSCs that were not evident in the parental fibroblasts, which may result from reprogramming. There were only small differences in the total numbers and types of variants among different reprogramming methods. Most importantly, a thorough genomic analysis showed that the variants were generally benign. We conclude that the process of reprogramming is unlikely to introduce variants that would make the cells inappropriate for therapy. It is feared that reprogramming may introduce DNA mutations. Here Bhutani et al. take three different reprogramming methods and using comparative whole genome analyses do identify nucleotide variations that are different in reprogrammed cells from the original fibroblasts, but none convey oncogenic potential.