CTCF counter-regulates cardiomyocyte development and maturation programs in the embryonic heart.
CTCF counter-regulates cardiomyocyte development and maturation programs in the embryonic heart.
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DOI：
10.1371/journal.pgen.1006985
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发表时间：
2017-08
期刊：
影响因子：
4.5
通讯作者：
Manzanares M
中科院分区：
文献类型：
--
作者：
Gomez-Velazquez M;Badia-Careaga C;Lechuga-Vieco AV;Nieto-Arellano R;Tena JJ;Rollan I;Alvarez A;Torroja C;Caceres EF;Roy AR;Galjart N;Delgado-Olguin P;Sanchez-Cabo F;Enriquez JA;Gomez-Skarmeta JL;Manzanares M
心脏祖细胞在发育早期被指定，并逐渐分化并成熟为功能齐全的心肌细胞。这个过程是由广泛研究的转录程序控制的。然而，协调此类项目从开发到成熟的进程的监管事件在很大程度上是未知的。在这里，我们表明基因组组织者 CTCF 对于心脏发生至关重要，并且它介导基因组相互作用以协调发育中的心脏中的心肌细胞分化和成熟。心脏祖细胞及其衍生物在发育过程中体内 Ctcf 失活导致严重的心脏缺陷，并在胚胎第 12.5 天死亡。 Ctcf 突变心脏的全基因组表达分析表明，心肌细胞成熟所需的控制线粒体功能和蛋白质产生的基因上调。然而，突变心肌细胞的线粒体不能正常成熟。相比之下，预测的心脏增强子附近的多个发育调节基因（包括 IrxA 簇中的基因）在 Ctcf 突变体中下调，表明 CTCF 通过促进增强子-启动子相互作用来促进心肌细胞分化。因此，CTCF 的丢失会破坏基因表达和染色质相互作用，如染色质构象捕获和深度测序所示。此外，CRISPR介导的IrxA簇内基因间CTCF位点的缺失会改变发育中心脏的基因表达。因此，CTCF 介导局部调节相互作用，以协调控制心脏发育过程中形态和功能转变的转录程序。在发育和分化过程中，在时间和空间上正确调节基因表达不仅需要转录输入，还需要染色质的特定结构。 CTCF 是一种 DNA 结合因子，通过与基因组中数以万计的位点结合，被认为对于这一过程至关重要。尽管近年来人们对 CTCF 在基因组组织中的作用有了一定的认识，但对其在体内的功能却知之甚少。为了解决这个问题，我们研究了基因删除 CTCF 对小鼠发育早期阶段心肌细胞分化的影响。令人惊讶的是，当 CTCF 被去除时，只有一小部分基因的表达发生变化。重要的是，失调的基因一方面控制着相反的遗传程序，负责发育和模式化，另一方面负责心肌细胞的成熟。这种不平衡导致线粒体故障和心脏模式基因的不正确表达，以及随后的胚胎致死。我们的结果表明，CTCF 对于维持全局基因组结构不是必需的，而是协调控制发育细胞和组织中表型转变的动态遗传程序。
Cardiac progenitors are specified early in development and progressively differentiate and mature into fully functional cardiomyocytes. This process is controlled by an extensively studied transcriptional program. However, the regulatory events coordinating the progression of such program from development to maturation are largely unknown. Here, we show that the genome organizer CTCF is essential for cardiogenesis and that it mediates genomic interactions to coordinate cardiomyocyte differentiation and maturation in the developing heart. Inactivation of Ctcf in cardiac progenitor cells and their derivatives in vivo during development caused severe cardiac defects and death at embryonic day 12.5. Genome wide expression analysis in Ctcf mutant hearts revealed that genes controlling mitochondrial function and protein production, required for cardiomyocyte maturation, were upregulated. However, mitochondria from mutant cardiomyocytes do not mature properly. In contrast, multiple development regulatory genes near predicted heart enhancers, including genes in the IrxA cluster, were downregulated in Ctcf mutants, suggesting that CTCF promotes cardiomyocyte differentiation by facilitating enhancer-promoter interactions. Accordingly, loss of CTCF disrupts gene expression and chromatin interactions as shown by chromatin conformation capture followed by deep sequencing. Furthermore, CRISPR-mediated deletion of an intergenic CTCF site within the IrxA cluster alters gene expression in the developing heart. Thus, CTCF mediates local regulatory interactions to coordinate transcriptional programs controlling transitions in morphology and function during heart development. Properly regulated gene expression in time and space during development and differentiation requires not only transcriptional inputs, but also specific structuring of the chromatin. CTCF is a DNA binding factor that is believed to be critical for this process through binding to tens of thousands of sites across the genome. Despite the knowledge gained in recent years on the role of CTCF in genome organization, its functions in vivo are poorly understood. To address this issue, we studied the effect of genetically deleting CTCF in differentiating cardiomyocytes at early stages of mouse development. Surprisingly only a fraction of genes change their expression when CTCF is removed. Importantly, misregulated genes control opposing genetic programs in charge of development and patterning on one hand, and cardiomyocyte maturation on the other. This imbalance leads to faulty mitochondria and incorrect expression of cardiac patterning genes, and subsequent embryonic lethality. Our results suggest that CTCF is not necessary for maintenance of global genome structure, but coordinates dynamic genetic programs controlling phenotypic transitions in developing cells and tissues.
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影响因子：
64.8
作者：
Jin, Fulai;Li, Yan;Dixon, Jesse R.;Selvaraj, Siddarth;Ye, Zhen;Lee, Ah Young;Yen, Chia-An;Schmitt, Anthony D.;Espinoza, Celso A.;Ren, Bing
通讯作者：
Ren, Bing
影响因子：
3.7
作者：
Beutner G;Eliseev RA;Porter GA Jr
通讯作者：
Porter GA Jr
影响因子：
64.5
作者：
Dowen JM;Fan ZP;Hnisz D;Ren G;Abraham BJ;Zhang LN;Weintraub AS;Schujiers J;Lee TI;Zhao K;Young RA
通讯作者：
Young RA
影响因子：
11.4
作者：
Heath, Helen;de Almeida, Claudia Ribeiro;Galjart, Niels
通讯作者：
Galjart, Niels
影响因子：
4.8
作者：
Fernández-Vizarra, E;López-Pérez, MJ;Enriquez, JA
通讯作者：
Enriquez, JA