Sin3a drives mesenchymal-to-epithelial transition through cooperating with Tet1 in somatic cell reprogramming.
Sin3a drives mesenchymal-to-epithelial transition through cooperating with Tet1 in somatic cell reprogramming.
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DOI：
10.1186/s13287-022-02707-4
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发表时间：
2022-01-24
影响因子：
7.5
通讯作者：
Kang J
中科院分区：
文献类型：
--
作者：
Feng J;Zhu F;Ye D;Zhang Q;Guo X;Du C;Kang J
识别新的调控因子和揭示体细胞重编程的机制将有助于诱导多能干细胞（iPSCs）的基础研究和临床应用。Sin 3a是一种多功能的转录调控因子，已被证实参与维持胚胎干细胞（embryonic stem cells，ESCs）的多能性，但Sin 3a在体细胞重编程中的作用尚不清楚。在体细胞重编程过程中，Sin 3a的RNA干扰是通过短发夹RNA实现的。通过碱性磷酸酶（AP）阳性菌落和Oct 4-GFP阳性菌落的数量评价重编程效率。进行RNA测序以鉴定Sin 3a敲低后受影响的生物学过程，并通过定量RT-PCR（qRT-PCR）、蛋白质印迹和流式细胞术进一步证实。免疫共沉淀法检测Sin 3a与Tet 1的相互作用。通过染色质免疫沉淀法测定Sin 3a和Tet 1在上皮基因启动子处的富集。此外，在基因位点的DNA甲基化模式进行了研究，通过羟甲基化DNA免疫沉淀。最后，还引入了破坏Sin 3a和Tet 1相互作用的Sin 3a突变体，以评估Sin 3a-Tet 1相互作用在间充质向上皮转化（MET）过程中的重要性。我们发现Sin 3a在OSKM诱导的重编程过程中逐渐增加，并且Sin 3a的敲低在重编程和iPSC产生的早期阶段显著损害MET。机制研究表明，Sin 3a招募Tet 1，以促进上皮基因启动子的羟甲基化。此外，破坏Sin 3a和Tet 1的相互作用显著阻断MET和iPSC的产生。我们的研究表明，Sin 3a是MET在早期重编程过程中的一种新型介导剂，其中Sin 3a作为表观遗传共激活因子，与Tet 1合作激活上皮程序并促进体细胞重编程的启动。这些发现突出了Sin 3a在MET过程中的重要性，并加深了我们对早期重编程的表观遗传调控网络的理解。在线版本包含补充材料，可通过10.1186/s13287-022-02707-4获得。
Identifying novel regulatory factors and uncovered mechanisms of somatic cell reprogramming will be helpful for basic research and clinical application of induced pluripotent stem cells (iPSCs). Sin3a, a multifunctional transcription regulator, has been proven to be involved in the maintenance of pluripotency in embryonic stem cells (ESCs), but the role of Sin3a in somatic cell reprogramming remains unclear. RNA interference of Sin3a during somatic cell reprogramming was realized by short hairpin RNAs. Reprogramming efficiency was evaluated by the number of alkaline phosphatase (AP)-positive colonies and Oct4-GFP-positive colonies. RNA sequencing was performed to identify the influenced biological processes after Sin3a knockdown and further confirmed by quantitative RT-PCR (qRT-PCR), western blotting and flow cytometry. The interaction between Sin3a and Tet1 was detected by coimmunoprecipitation. The enrichment of Sin3a and Tet1 at the epithelial gene promoters was measured by chromatin immunoprecipitation. Furthermore, DNA methylation patterns at the gene loci were investigated by hydroxymethylated DNA immunoprecipitation. Finally, Sin3a mutants that disrupt the interaction of Sin3a and Tet1 were also introduced to assess the importance of the Sin3a–Tet1 interaction during the mesenchymal-to-epithelial transition (MET) process. We found that Sin3a was gradually increased during OSKM-induced reprogramming and that knockdown of Sin3a significantly impaired MET at the early stage of reprogramming and iPSC generation. Mechanistic studies showed that Sin3a recruited Tet1 to facilitate the hydroxymethylation of epithelial gene promoters. Moreover, disrupting the interaction of Sin3a and Tet1 significantly blocked MET and iPSC generation. Our studies revealed that Sin3a was a novel mediator of MET during early reprogramming, where Sin3a functioned as an epigenetic coactivator, cooperating with Tet1 to activate the epithelial program and promote the initiation of somatic cell reprogramming. These findings highlight the importance of Sin3a in the MET process and deepen our understanding of the epigenetic regulatory network of early reprogramming. The online version contains supplementary material available at 10.1186/s13287-022-02707-4.
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DOI：
10.1016/j.bbagrm.2009.05.007
发表时间：
2009-06
期刊：
Biochimica et biophysica acta
影响因子：
--
作者：
Grzenda A;Lomberk G;Zhang JS;Urrutia R
通讯作者：
Urrutia R
影响因子：
2.5
作者：
Schwab, Renate H. M.;Amin, Nancy;Vincan, Elizabeth
通讯作者：
Vincan, Elizabeth
影响因子：
9.7
作者：
Lili, Loukia N.;Huang, Andrew D.;McDonald, John F.
通讯作者：
McDonald, John F.
影响因子：
4.6
作者：
Chandru A;Bate N;Vuister GW;Cowley SM
通讯作者：
Cowley SM
影响因子：
14.8
作者：
Lee, Tong Ihn;Johnstone, Sarah E.;Young, Richard A.
通讯作者：
Young, Richard A.