Epigenetic Modifiers Facilitate Induction and Pluripotency of Porcine iPSCs.
Epigenetic Modifiers Facilitate Induction and Pluripotency of Porcine iPSCs.
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DOI：
10.1016/j.stemcr.2016.11.013
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发表时间：
2017-01-10
影响因子：
5.9
通讯作者：
Liu L
中科院分区：
文献类型：
--
作者：
Mao J;Zhang Q;Deng W;Wang H;Liu K;Fu H;Zhao Q;Wang X;Liu L
外源基因的不充分沉默是传统多能转录因子（OSKM）对猪诱导多能干细胞（piPSC）完成表观遗传重编程的主要障碍。我们测试了以下假设：通过活性 DNA 或组蛋白去甲基化或通过抑制组蛋白脱乙酰酶进行表观遗传修饰将增强 piPSC 中的重编程和外源基因沉默。与 OSKM 对照相比，OSKM 与表观遗传因子（特别是 10-11 易位（Tet1 或 Tet3）或赖氨酸 (K) 特异性去甲基酶 3A (Kdm3a)）相结合诱导的 piPSC 表达更高水平的 Rex1 和代表幼稚状态的其他基因，并表现出更开放的染色质状态。 Tet1还提高了分化能力。使用组蛋白脱乙酰酶 (HDACi)、NaB、TSA 或 VPA 抑制剂进行转化，进一步增加 Rex1 表达，同时降低外源基因的表达。由 Tet1+OSKM 诱导并随后用 HDACi 转化的 piPSC 显示出高多能性。表观遗传修饰剂共同增强 piPSC 的生成并减少其对外源基因的依赖。表观遗传修饰剂促进猪 iPSC 的诱导和质量 Tet1、Tet3 或 Kdm3a 增加与 Rex1 相关的幼稚多能性网络 与细胞质 Rex1 不同，Rex1 的核表达与高多能性 HDAC 抑制剂相关，进一步激活 Rex1 并减少对外源基因的依赖。可以显着增强常规山中因子对猪 iPSC 的诱导和多能性，如细胞核中 Rex1 表达水平及其多能性网络的增加以及畸胎瘤形成试验所证明。此外，通过用HDAC抑制剂进一步治疗，外源基因的表达减少。
Inadequate silencing of exogenous genes represents a major obstacle to complete epigenetic reprogramming of porcine-induced pluripotent stem cells (piPSCs) by conventional pluripotency transcription factors (OSKM). We tested the hypothesis that epigenetic modification by active DNA or histone demethylation or by inhibition of histone deacetylase would enhance reprogramming and exogenous gene silencing in piPSCs. piPSCs induced by OSKM in combination with epigenetic factors, specifically Ten-Eleven Translocation (Tet1 or Tet3) or lysine (K)-specific demethylase 3A (Kdm3a), expressed higher levels of Rex1 and other genes representing naive state and exhibited more open chromatin status, compared with those of OSKM controls. Tet1 also improved differentiation capacity. Conversion with inhibitors of histone deacetylases (HDACi), NaB, TSA, or VPA, further increased Rex1 expression, while decreasing expression of exogenous genes. piPSCs induced by Tet1+OSKM followed by conversion with HDACi show high pluripotency. Together, epigenetic modifiers enhance generation of piPSCs and reduce their reliance on exogenous genes. Epigenetic modifiers facilitate induction and quality of porcine iPSCs Tet1, Tet3, or Kdm3a increases naive pluripotency network in association with Rex1 Unlike cytoplasmic Rex1, nuclear expression of Rex1 is associated with high pluripotency HDAC inhibitors further activate Rex1 and reduce reliance on the exogenous genes In this article, Liu and colleagues show that epigenetic modifiers, Tet1, Tet3, or Kdm3a can significantly enhance induction and pluripotency of porcine iPSCs by conventional Yamanaka factors, as demonstrated by increased expression levels of Rex1 in the nuclei and its pluripotency network, and by teratoma formation test. Moreover, expression of exogenous genes is decreased by further treatment with HDAC inhibitor.