FOXF1转录因子通过调节内皮细胞中的VEGF信号传导来参与胚胎血管系统的形成。

FOXF1 transcription factor is required for formation of embryonic vasculature by regulating VEGF signaling in endothelial cells.

作者信息

Ren Xiaomeng, Ustiyan Vladimir, Pradhan Arun, Cai Yuqi, Havrilak Jamie A, Bolte Craig S, Shannon John M, Kalin Tanya V, Kalinichenko Vladimir V

机构信息

From the Divisons of Pulmonary Biology (X.R., V.U., A.P., Y.C., J.A.H., C.S.B., J.M.S., T.V.K., V.V.K.) and Developmental Biology (V.V.K.), Perinatal Institute, Cincinnati Children's Research Foundation, OH.

出版信息

Circ Res. 2014 Sep 26;115(8):709-20. doi: 10.1161/CIRCRESAHA.115.304382. Epub 2014 Aug 4.

DOI:10.1161/CIRCRESAHA.115.304382

PMID:25091710

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4810682/

Abstract

RATIONALE

Inactivating mutations in the Forkhead Box transcription factor F1 (FOXF1) gene locus are frequently found in patients with alveolar capillary dysplasia with misalignment of pulmonary veins, a lethal congenital disorder, which is characterized by severe abnormalities in the respiratory, cardiovascular, and gastrointestinal systems. In mice, haploinsufficiency of the Foxf1 gene causes alveolar capillary dysplasia and developmental defects in lung, intestinal, and gall bladder morphogenesis.

OBJECTIVE

Although FOXF1 is expressed in multiple mesenchyme-derived cell types, cellular origins and molecular mechanisms of developmental abnormalities in FOXF1-deficient mice and patients with alveolar capillary dysplasia with misalignment of pulmonary veins remain uncharacterized because of lack of mouse models with cell-restricted inactivation of the Foxf1 gene. In the present study, the role of FOXF1 in endothelial cells was examined using a conditional knockout approach.

METHODS AND RESULTS

A novel mouse line harboring Foxf1-floxed alleles was generated by homologous recombination. Tie2-Cre and Pdgfb-CreER transgenes were used to delete Foxf1 from endothelial cells. FOXF1-deficient embryos exhibited embryonic lethality, growth retardation, polyhydramnios, cardiac ventricular hypoplasia, and vascular abnormalities in the lung, placenta, yolk sac, and retina. Deletion of FOXF1 from endothelial cells reduced endothelial proliferation, increased apoptosis, inhibited vascular endothelial growth factor signaling, and decreased expression of endothelial genes critical for vascular development, including vascular endothelial growth factor receptors Flt1 and Flk1, Pdgfb, Pecam1, CD34, integrin β3, ephrin B2, Tie2, and the noncoding RNA Fendrr. Chromatin immunoprecipitation assay demonstrated that Flt1, Flk1, Pdgfb, Pecam1, and Tie2 genes are direct transcriptional targets of FOXF1.

CONCLUSIONS

FOXF1 is required for the formation of embryonic vasculature by regulating endothelial genes critical for vascular development and vascular endothelial growth factor signaling.

摘要

原理

叉头框转录因子F1（FOXF1）基因位点的失活突变在患有肺静脉错位的肺泡毛细血管发育不良患者中经常被发现，这是一种致命的先天性疾病，其特征是呼吸、心血管和胃肠道系统存在严重异常。在小鼠中，Foxf1基因的单倍剂量不足会导致肺泡毛细血管发育不良以及肺、肠和胆囊形态发生的发育缺陷。

目的

尽管FOXF1在多种间充质来源的细胞类型中表达，但由于缺乏Foxf1基因细胞限制性失活的小鼠模型，FOXF1缺陷小鼠和患有肺静脉错位的肺泡毛细血管发育不良患者发育异常的细胞起源和分子机制仍未明确。在本研究中，使用条件性敲除方法研究了FOXF1在内皮细胞中的作用。

方法与结果

通过同源重组产生了一种携带Foxf1基因条件性敲除等位基因的新型小鼠品系。使用Tie2-Cre和Pdgfb-CreER转基因从内皮细胞中删除Foxf1。FOXF1缺陷的胚胎表现出胚胎致死、生长迟缓、羊水过多、心室发育不全以及肺、胎盘、卵黄囊和视网膜的血管异常。从内皮细胞中删除FOXF1会降低内皮细胞增殖、增加细胞凋亡、抑制血管内皮生长因子信号传导，并降低对血管发育至关重要的内皮基因的表达，包括血管内皮生长因子受体Flt1和Flk1、Pdgfb、Pecam1、CD34、整合素β3、ephrin B2、Tie2以及非编码RNA Fendrr。染色质免疫沉淀试验表明，Flt1、Flk1、Pdgfb、Pecam1和Tie2基因是FOXF1的直接转录靶点。