Spatial transcriptional profile of the chick and mouse endocardial cushions identify novel regulators of endocardial EMT in vitro.
Spatial transcriptional profile of the chick and mouse endocardial cushions identify novel regulators of endocardial EMT in vitro.
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雏鸡和小鼠心内膜垫的空间转录曲线在体外鉴定了内膜EMT的新调节剂。
DOI：
10.1016/j.yjmcc.2013.03.016
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发表时间：
2013-06
影响因子：
5
通讯作者：
Barnett JV
中科院分区：
文献类型：
--
作者：
DeLaughter DM;Christodoulou DC;Robinson JY;Seidman CE;Baldwin HS;Seidman JG;Barnett JV
瓣膜间质细胞（VIC）是先天性和成人心脏病的常见基质，但在早期瓣膜发生期间控制其形成的信号传导机制尚未完全了解。我们开发了一个公正的策略，以确定重要的基因在内皮细胞上皮间质转化（EMT）使用的空间转录谱。覆盖在房室管（AVC）和流出道（OFT）垫上的内皮细胞经历EMT以产生VIC。对从发育相当阶段的鸡和小鼠胚胎（鸡HH 18;小鼠E11.0）分离的AVC、OFT和心室（VEN）之间的基因表达进行RNA测序（RNA-seq）分析。EMT发生在AVC和OFT缓冲垫中，但此时未发生VEN。鸡（n=1）中的198个基因和小鼠（n=2）中的105个基因在垫中富集2倍。从富含TGF β的基因列表生成的基因调控网络（GRN）确认TGFβ为节点，并将NF-κB鉴定为潜在节点。为了揭示以前未被识别的EMT调节因子，选择了4个候选基因Hapln 1、Id 1、Foxp 2和Meis 2以及候选途径NF-κB。通过原位杂交证实每个基因的体内空间表达，并且通过胶原凝胶测定中的siRNA敲低确定每个基因在内皮细胞EMT中的功能作用。我们的空间-转录分析策略产生了反映系统已知生物学的基因列表。进一步的分析准确地鉴定和验证了以前未被识别的新的候选基因和NF-κB通路作为体外内皮细胞EMT的调节因子。
Valvular Interstitial Cells (VICs) are a common substrate for congenital and adult heart disease yet the signaling mechanisms governing their formation during early valvulogenesis are incompletely understood. We developed an unbiased strategy to identify genes important in endocardial epithelial-to-mesenchymal transformation (EMT) using a spatial transcriptional profile. Endocardial cells overlaying the cushions of the atrioventricular canal (AVC) and outflow tract (OFT) undergo an EMT to yield VICs. RNA sequencing (RNA-seq) analysis of gene expression between AVC, OFT, and ventricles (VEN) isolated from chick and mouse embryos at comparable stages of development (chick HH18; mouse E11.0) was performed. EMT occurs in the AVC and OFT cushions, but not VEN at this time. 198 genes in the chick (n=1) and 105 genes in the mouse (n=2) were enriched 2-fold in the cushions. Gene regulatory networks (GRN) generated from cushion-enriched gene lists confirmed TGFβ as a nodal point and identified NF-κB as a potential node. To reveal previously unrecognized regulators of EMT four candidate genes, Hapln1, Id1, Foxp2, and Meis2, and a candidate pathway, NF-κB, were selected. In vivo spatial expression of each gene was confirmed by in situ hybridization and a functional role for each in endocardial EMT was determined by siRNA knockdown in a collagen gel assay. Our spatial-transcriptional profiling strategy yielded gene lists which reflected the known biology of the system. Further analysis accurately identified and validated previously unrecognized novel candidate genes and the NF-κB pathway as regulators of endocardial cell EMT in vitro.
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影响因子：
2.5
作者：
Azhar, Mohamad;Runyan, Raymond B.;Gard, Connie;Sanford, L. Philip;Miller, Marian L.;Andringa, Anastasia;Pawlowski, Sharon;Rajan, Sudarsan;Doetschman, Thomas
通讯作者：
Doetschman, Thomas
影响因子：
16
作者：
Capdevila, J;Tsukui, T;Belmonte, JCI
通讯作者：
Belmonte, JCI
影响因子：
2.7
作者：
Camenisch, TD;Molin, DGM;Klewer, SE
通讯作者：
Klewer, SE
影响因子：
64.8
作者：
BEG, AA;SHA, WC;BALTIMORE, D
通讯作者：
BALTIMORE, D
影响因子：
4.6
作者：
Chakraborty, Santanu;Cheek, Jonathan;Yutzey, Katherine E.
通讯作者：
Yutzey, Katherine E.