A Loss of Function Screen of Epigenetic Modifiers and Splicing Factors during Early Stage of Cardiac Reprogramming.
A Loss of Function Screen of Epigenetic Modifiers and Splicing Factors during Early Stage of Cardiac Reprogramming.
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DOI：
10.1155/2018/3814747
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发表时间：
2018
影响因子：
4.3
通讯作者：
Qian L
中科院分区：
文献类型：
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作者：
Zhou Y;Alimohamadi S;Wang L;Liu Z;Wall JB;Yin C;Liu J;Qian L
心脏成纤维细胞（CFs）直接重编程为诱导心肌细胞（iCMs）是一种新兴的有前途的心脏再生、疾病建模和药物发现方法。然而，由于低重编程效率和大量未知的潜在分子机制，其潜力受到极大限制。我们之前已经筛选并确定了与iCM重编程过程中组蛋白修饰相关的表观遗传因素。在这里，我们使用shrna靶向另一组参与染色质重塑的表观遗传因子和RNA剪接因子，以进一步鉴定直接心脏重编程的抑制剂和促进因子。RNA剪接因子Sf3a1或Sf3b1的敲低显著降低心脏标志物阳性iCMs的百分比和总数，同时基因表达普遍抑制。去除另一种RNA剪接因子Zrsr2可促进CFs和小鼠胚胎成纤维细胞（MEFs）在蛋白质和mRNA水平上获得CM分子特征。此外，用靶向Bcor复合物超家族组分或Stag2（黏结蛋白复合物组分）的shrna处理的cf和MEFs中，重编程效率一致提高。因此，我们的工作揭示了一些额外的表观遗传和剪接因子，这些因子要么抑制iCM重编程，要么是iCM重编程所必需的，并强调了表观遗传调控和RNA剪接过程在细胞命运转化中的重要性。
Direct reprogramming of cardiac fibroblasts (CFs) to induced cardiomyocytes (iCMs) is a newly emerged promising approach for cardiac regeneration, disease modeling, and drug discovery. However, its potential has been drastically limited due to the low reprogramming efficiency and largely unknown underlying molecular mechanisms. We have previously screened and identified epigenetic factors related to histone modification during iCM reprogramming. Here, we used shRNAs targeting an additional battery of epigenetic factors involved in chromatin remodeling and RNA splicing factors to further identify inhibitors and facilitators of direct cardiac reprogramming. Knockdown of RNA splicing factors Sf3a1 or Sf3b1 significantly reduced the percentage and total number of cardiac marker positive iCMs accompanied with generally repressed gene expression. Removal of another RNA splicing factor Zrsr2 promoted the acquisition of CM molecular features in CFs and mouse embryonic fibroblasts (MEFs) at both protein and mRNA levels. Moreover, a consistent increase of reprogramming efficiency was observed in CFs and MEFs treated with shRNAs targeting Bcor (component of BCOR complex superfamily) or Stag2 (component of cohesin complex). Our work thus reveals several additional epigenetic and splicing factors that are either inhibitory to or required for iCM reprogramming and highlights the importance of epigenetic regulation and RNA splicing process during cell fate conversion.
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影响因子：
1.2
作者：
Liu Z;Chen O;Zheng M;Wang L;Zhou Y;Yin C;Liu J;Qian L
通讯作者：
Qian L
影响因子：
64.5
作者：
Goldberg, Aaron D.;Allis, C. David;Bernstein, Emily
通讯作者：
Bernstein, Emily
影响因子：
4.3
作者：
Gupta, Preksha;Lavagnolli, Thais;Merkenschlager, Matthias
通讯作者：
Merkenschlager, Matthias
影响因子：
4
作者：
Luna-Zurita, Luis;Bruneau, Benoit G.
通讯作者：
Bruneau, Benoit G.
影响因子：
64.8
作者：
通讯作者：
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