Reprogramming triggers endogenous L1 and Alu retrotransposition in human induced pluripotent stem cells.
Reprogramming triggers endogenous L1 and Alu retrotransposition in human induced pluripotent stem cells.
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DOI：
10.1038/ncomms10286
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发表时间：
2016-01-08
影响因子：
16.6
通讯作者：
Schumann GG
中科院分区：
文献类型：
--
作者：
Klawitter S;Fuchs NV;Upton KR;Muñoz-Lopez M;Shukla R;Wang J;Garcia-Cañadas M;Lopez-Ruiz C;Gerhardt DJ;Sebe A;Grabundzija I;Merkert S;Gerdes P;Pulgarin JA;Bock A;Held U;Witthuhn A;Haase A;Sarkadi B;Löwer J;Wolvetang EJ;Martin U;Ivics Z;Izsvák Z;Garcia-Perez JL;Faulkner GJ;Schumann GG
人诱导多能干细胞(HiPSCs)具有无限增殖能力，并能在体外分化为三个初级生殖层的衍生体。Wissing和他的同事已经报告了在hiPSC衍生过程中发生的遗传和表观遗传学异常，包括动员工程化LINE-1(L1)反转录转座子。然而，内源性逆转录转位在HiPSCs中的发生率和功能影响尚不清楚。在这里，我们应用反转录转座子捕获测序对8个HiPSC系和3个人胚胎干细胞(HESC)系进行了测序，揭示了在重编程和多能干细胞培养过程中内源性L1、Alu和Sine-VNTR-Alu(SVA)的动员。令人惊讶的是，在多能干细胞中表达的蛋白质编码基因中，有4/7的L1插入是全长的，6/11发生了逆转录转座事件。我们进一步证明，在HiPSC培养过程中，CADPS2基因中的L1内含子插入并破坏了CADPS2的表达。这些实验阐明了hPSCs和hESCs中的内源性逆转座及其潜在后果。已发现遗传和表观遗传学异常是由于将分化的细胞重新编程为人类诱导的多能干细胞(HiPSCs)造成的。在这里，Klawitter等人。在重编程过程中识别内源性L1、Alu和SVA动员，强调HiPSC中插入突变剂的风险。
Human induced pluripotent stem cells (hiPSCs) are capable of unlimited proliferation and can differentiate in vitro to generate derivatives of the three primary germ layers. Genetic and epigenetic abnormalities have been reported by Wissing and colleagues to occur during hiPSC derivation, including mobilization of engineered LINE-1 (L1) retrotransposons. However, incidence and functional impact of endogenous retrotransposition in hiPSCs are yet to be established. Here we apply retrotransposon capture sequencing to eight hiPSC lines and three human embryonic stem cell (hESC) lines, revealing endogenous L1, Alu and SINE-VNTR-Alu (SVA) mobilization during reprogramming and pluripotent stem cell cultivation. Surprisingly, 4/7 de novo L1 insertions are full length and 6/11 retrotransposition events occurred in protein-coding genes expressed in pluripotent stem cells. We further demonstrate that an intronic L1 insertion in the CADPS2 gene is acquired during hiPSC cultivation and disrupts CADPS2 expression. These experiments elucidate endogenous retrotransposition, and its potential consequences, in hiPSCs and hESCs. Genetic and epigenetic abnormalities have been found to result from reprogramming of differentiated cells into human induced pluripotent stem cells (hiPSCs). Here, Klawitter et al. identify endogenous L1, Alu and SVA mobilization during reprogramming, highlighting the risk of insertional mutagens in hiPSCs.