FOXF1 transcription factor promotes lung regeneration after partial pneumonectomy

Abstract

FOXF1, a member of the forkhead box family of transcription factors, has been previously shown to be critical for lung development, homeostasis, and injury responses. However, the role of FOXF1 in lung regeneration is unknown. Herein, we performed partial pneumonectomy, a model of lung regeneration, in mice lacking one Foxf1 allele in endothelial cells (PDGFb-iCre/Foxf1 fl/+ mice). Endothelial cell proliferation was significantly reduced in regenerating lungs from mice deficient for endothelial Foxf1. Decreased endothelial proliferation was associated with delayed lung regeneration as shown by reduced respiratory volume in Foxf1-deficient lungs. FACS-sorted endothelial cells isolated from regenerating PDGFb-iCre/Foxf1 fl/+ and control lungs were used for RNAseq analysis to identify FOXF1 target genes. Foxf1 deficiency altered expression of numerous genes including those regulating extracellular matrix remodeling (Timp3, Adamts9) and cell cycle progression (Cdkn1a, Cdkn2b, Cenpj, Tubb4a), which are critical for lung regeneration. Deletion of Foxf1 increased Timp3 mRNA and protein, decreasing MMP14 activity in regenerating lungs. ChIPseq analysis for FOXF1 and histone methylation marks identified DNA regulatory regions within the Cd44, Cdkn1a, and Cdkn2b genes, indicating they are direct FOXF1 targets. Thus FOXF1 stimulates lung regeneration following partial pneumonectomy via direct transcriptional regulation of genes critical for extracellular matrix remodeling and cell cycle progression.

The forkhead box F1 transcription factor inhibits collagen deposition and accumulation of myofibroblasts during liver fibrosis

“…Desmin and αSMA were both increased in αSMACreER;Foxf1 −/− livers; however, there were no differences in the number of proliferating cells between Foxf1 fl/fl and αSMACreER;Foxf1 −/− livers. Previously, FOXF1 has been shown to stimulate cell proliferation in lung endothelial cells (Ren et al, 2014; Bolte et al, 2017) and in rhabdomyosarcoma tumor cells (Milewski et al, 2017). Surprisingly, we found that deletion of Foxf1 from MFs does not affect their proliferation during liver fibrogenesis.…”

“…Protein extracts were isolated using cell lysis buffer as previously described (Pradhan et al, 2016) and used for either western blot analysis with Pierce ECL western blotting substrate (Thermo Fisher Scientific) or gel zymography (NOVEX) according to the manufacturer’s protocols. The following antibodies were used for protein blots: FOXF1 (1:1000, R&D Systems) (Bolte et al, 2017; Black et al, 2018; Ren et al, 2014), ACTIN (1:2000; Santa Cruz Biotechnology) (Pradhan et al, 2016), FOXM1 (1:3000; Santa Cruz Biotechnology) (Pradhan et al, 2016; Bolte et al, 2011), CCND1 (1:1000; Cell Signaling Technology) (Milewski et al, 2017). Protein band intensities were determined by ImageJ software and were normalized to ACTIN.…”

Functional CRISPR screen identifies AP1-associated enhancer regulating FOXF1 to modulate oncogene-induced senescence

“…FOXA1 has been reported to promote senescence via activation of p16 INK4a [ 33 ] and FOXO4 inhibition induces p53 nuclear exclusion, which results in apoptosis of senescent cells [ 34 ]. The functions of FOXF1 remain to be determined, yet recent studies have implicated its role in lung regeneration by targeting genes of ECM and cell cycle progression [ 35 ], as well as promoting prostate cancer growth via the MAPK pathway [ 36 ]. To date, there has been no evidence of any connections between AP1, FOXF1, and OIS, possibly due to regulation via enhancers, rather than proximal promoters, as proposed in this study.…”

FOXF1 Induces Epithelial-Mesenchymal Transition in Colorectal Cancer Metastasis by Transcriptionally Activating SNAI1

“…In the present study, the up-regulation of FOXF1 could induce EMT in CRC, but the specific molecular mechanisms by which FOXF1 induces EMT require further discussion. As a transcription factor, FOXF1 binds to the promoter of the target genes to regulate gene expression and the activity of multiple signaling pathways that play an important role in physiological or pathological processes [10] , [11] , [13] , [43] , suggesting that FOXF1 might directly regulate master EMT-related genes to induce EMT in CRC. The SNAI1 protein has been identified as a master transcription regulator of EMT [44] .…”