PDF(Pubmed)
Abstract：
Mutations in the FOXF1 gene, a key transcriptional regulator of pulmonary vascular development, cause Alveolar Capillary Dysplasia with Misalignment of Pulmonary Veins, a lethal lung disease affecting newborns and infants. Identification of new FOXF1 upstream regulatory elements is critical to explain why frequent non-coding FOXF1 deletions are linked to the disease. Herein, we use multiome single-nuclei RNA and ATAC sequencing of mouse and human patient lungs to identify four conserved endothelial and mesenchymal FOXF1 enhancers. We demonstrate that endothelial FOXF1 enhancers are autoactivated, whereas mesenchymal FOXF1 enhancers are regulated by EBF1 and GLI1. The cell-specificity of FOXF1 enhancers is validated by disrupting these enhancers in mouse embryonic stem cells using CRISPR/Cpf1 genome editing followed by lineage-tracing of mutant embryonic stem cells in mouse embryos using blastocyst complementation. This study resolves an important clinical question why frequent non-coding FOXF1 deletions that interfere with endothelial and mesenchymal enhancers can lead to the disease.
摘要：
FOXF1基因突变,肺血管发育的关键转录调节因子，导致肺泡毛细血管发育不良伴肺静脉错位，一种影响新生儿和婴儿的致命肺病。鉴定新的FOXF1上游调控元件对于解释为什么频繁的非编码FOXF1缺失与疾病相关至关重要。在这里，我们使用小鼠和人类患者肺的多体单核RNA和ATAC测序来鉴定四种保守的内皮和间充质FOXF1增强子。我们证明内皮FOXF1增强子是自动激活的，而间充质FOXF1增强子受EBF1和GLI1调节。通过使用CRISPR/Cpf1基因组编辑破坏小鼠胚胎干细胞中的这些增强子，然后使用胚泡互补对小鼠胚胎中的突变胚胎干细胞进行谱系追踪，来验证FOXF1增强子的细胞特异性。这项研究解决了一个重要的临床问题，为什么频繁的非编码FOXF1缺失会干扰内皮和间充质增强剂导致疾病。
