Xist deficiency and disorders of X-inactivation in rabbit embryonic stem cells can be rescued by transcription-factor-mediated conversion.
Xist deficiency and disorders of X-inactivation in rabbit embryonic stem cells can be rescued by transcription-factor-mediated conversion.
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DOI：
10.1089/scd.2014.0011
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发表时间：
2014-05
影响因子：
4
通讯作者：
Yonghua Jiang;Zhaohui Kou;Tong Wu;W. An;R. Zhou;Hong Wang;Yawei Gao;Shaorong Gao
中科院分区：
文献类型：
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作者：
Yonghua Jiang;Zhaohui Kou;Tong Wu;W. An;R. Zhou;Hong Wang;Yawei Gao;Shaorong Gao
失活X染色体上X失活特异性转录本（XIST）的缺乏会影响女性人类胚胎干细胞（hESC）和人类诱导多能干细胞（hiPSC）的行为，并且随着这些细胞的持续传代，可能会发生进一步的染色体侵蚀。然而，X染色体不稳定性尚未在其他物种中被发现。在本研究中，我们研究了三个雌性兔ESC（rbESC）系，发现其中两个正常表达Xist并获得Xist RNA涂层和H3K27me3焦点，因此将其定义为Xi（Xist）Xa。有趣的是，第三个雌性 rbESC 系在 ESC 维持和分化过程中缺乏 Xist 表达。该品系在早期传代中显示 H3K27me3 焦点，但没有 Xist RNA 涂层，因此被定义为 Xi(w/oXist)Xa。与 Xi(w/oXist)Xa hESC 或 hiPSC 类似，Xi(w/oXist)Xa rbESC 失去 H3K27me3 并在传代时经历 Xi 侵蚀 (Xe)。此外，Xist缺陷的rbESC还表现出分化能力受损和癌症相关基因上调。通过在优化培养条件下在 Xist 缺陷的 rbESC 中过表达 OCT4、SOX2、KLF4 和 c-MYC，我们成功获得了小鼠 ESC 样（mESC 样）细胞。 mESC 样 rbESC 表现出圆顶形集落形态、LIF/STAT3 依赖性途径的激活以及无序 X 染色体的转化。重要的是，有缺陷的分化潜能也得到了极大的提高。我们的数据表明，X 染色体失活的变化发生在 rbESC 的早期传代中；因此，Xi 紊乱在物种间是保守的，并且使用适当的表观遗传重编程和培养条件是可逆的。这些发现可能对于未来衍生完全多能 rbESC 或兔 iPSC (rbiPSC) 的努力非常有用。
The deficiency of X-inactive specific transcript (XIST) on the inactive X chromosome affects the behavior of female human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs), and further chromosomal erosion can occur with continued passaging of these cells. However, X chromosome instability has not been identified in other species. In the present study, we investigated three female rabbit ESC (rbESC) lines and found that two of them expressed Xist normally and obtained both Xist RNA coating and H3K27me3 foci, thus defined as Xi(Xist)Xa. Interestingly, the third female rbESC line lacked Xist expression during ESC maintenance and differentiation. This line showed H3K27me3 foci but no Xist RNA coating in the early passages and was thus defined as Xi(w/oXist)Xa. Similar to Xi(w/oXist)Xa hESCs or hiPSCs, Xi(w/oXist)Xa rbESCs lose H3K27me3 and undergo Xi erosion (Xe) with passaging. Moreover, Xist-deficient rbESCs also exhibit impaired differentiation ability and upregulation of cancer-related genes. By overexpressing OCT4, SOX2, KLF4, and c-MYC in Xist-deficient rbESCs under optimized culture conditions, we successfully obtained mouse ESC-like (mESC-like) cells. The mESC-like rbESCs displayed dome-shaped colony morphology, activation of the LIF/STAT3-dependent pathway, and conversion of disordered X chromosome. Importantly, the defective differentiation potential was also greatly improved. Our data demonstrate that variations in X chromosome inactivation occur in early passage of rbESCs; thus, Xi disorders are conserved across species and are reversible using the proper epigenetic reprogramming and culture conditions. These findings may be very useful for future efforts toward deriving fully pluripotent rbESCs or rabbit iPSCs (rbiPSCs).