FOXK1 regulates Wnt signalling to promote cardiogenesis.
FOXK1 regulates Wnt signalling to promote cardiogenesis.
复制标题
FOXK1 调节 Wnt 信号传导以促进心脏发生。
DOI：
10.1093/cvr/cvad054
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发表时间：
2023
影响因子：
10.8
通讯作者：
Garry,DanielJ
中科院分区：
文献类型：
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作者：
Sierra-Pagan,JavierE;Dsouza,Nikita;Das,Satyabrata;Larson,ThijsA;Sorensen,JacobR;Ma,Xiao;Stan,Patricia;Wanberg,ErikJ;Shi,Xiaozhong;Garry,MaryG;Gong,Wuming;Garry,DanielJ
目的先天性心脏病（CHD）是最常见的遗传性先天缺陷，具有相当高的发病率和死亡率。我们专注于破译控制心脏祖细胞和心脏发生的关键调节因子。FOXK1是一种叉头/翼状螺旋转录因子，已知用于调节细胞周期动力学，并且仅限于中胚层祖细胞、体细胞和心脏。在本研究中，我们确定了FOXK1在心血管发育过程中的重要作用。方法与结果利用小鼠胚状体系统将对照和foxk1ko胚胎干细胞分化为中胚层细胞、心脏祖细胞和成熟心脏细胞。通过流式细胞术、免疫组织化学、心脏跳动、转录和染色质免疫沉淀定量聚合酶链反应测定、大量RNA测序（RNAseq）和转座酶可及染色质测序（ATACseq）分析，FOXK1被观察到是心脏发生的重要调节因子。流式细胞术分析显示foxk1ko胚胎样体（EBs）的心脏发生受到干扰。在两个发育阶段的大量RNAseq分析显示，与对照EBs相比，心脏分子程序inFoxk1KO EBs显着减少。在EB分化过程中的ATACseq分析表明，与foxk1ko EBs相比，对照EBs中已知的重要心脏发生调节因子附近的染色质景观显着放松。此外，我们通过检测心肌肌钙蛋白T表达和心肌收缩力证明，在缺乏FOXK1的情况下，心脏分化明显受损。我们证明FOXK1通过抑制Wnt/β-catenin信号通路从而促进分化是心脏发生的重要调节因子。结论FOXK1是心血管发育的重要转录和表观遗传调控因子。在机制上，FOXK1抑制Wnt信号以促进心脏祖细胞的发育。
AimsCongenital heart disease (CHD) is the most common genetic birth defect, which has considerable morbidity and mortality. We focused on deciphering key regulators that govern cardiac progenitors and cardiogenesis. FOXK1 is aforkhead/winged helixtranscription factor known to regulate cell cycle kinetics and is restricted to mesodermal progenitors, somites, and heart. In the present study, we define an essential role for FOXK1 during cardiovascular development.Methods and resultsWe used the mouse embryoid body system to differentiate control andFoxk1KO embryonic stem cells into mesodermal, cardiac progenitor cells and mature cardiac cells. Using flow cytometry, immunohistochemistry, cardiac beating, transcriptional and chromatin immunoprecipitation quantitative polymerase chain reaction assays, bulk RNA sequencing (RNAseq) and assay for transposase-accessible chromatin using sequencing (ATACseq) analyses, FOXK1 was observed to be an important regulator of cardiogenesis. Flow cytometry analyses revealed perturbed cardiogenesis inFoxk1KO embryoid bodies (EBs). Bulk RNAseq analysis at two developmental stages showed a significant reduction of the cardiac molecular program inFoxk1KO EBs compared to the control EBs. ATACseq analysis during EB differentiation demonstrated that the chromatin landscape nearby known important regulators of cardiogenesis was significantly relaxed in control EBs compared toFoxk1KO EBs. Furthermore, we demonstrated that in the absence of FOXK1, cardiac differentiation was markedly impaired by assaying for cardiac Troponin T expression and cardiac contractility. We demonstrate that FOXK1 is an important regulator of cardiogenesis by repressing the Wnt/β-catenin signalling pathway and thereby promoting differentiation.ConclusionThese results identify FOXK1 as an essential transcriptional and epigenetic regulator of cardiovascular development. Mechanistically, FOXK1 represses Wnt signalling to promote the development of cardiac progenitor cells.