title: FOXF1 transcription factor is required for formation of embryonic vasculature by regulating VEGF signaling in endothelial cells.
abstract: BACKGROUND
  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.
  
  
  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.
authors.affiliation: 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.
authors.forename: Xiaomeng
authors.initials: X
authors.lastname: Ren
authors.affiliation: 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.
authors.forename: Vladimir
authors.initials: V
authors.lastname: Ustiyan
authors.affiliation: 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.
authors.forename: Arun
authors.initials: A
authors.lastname: Pradhan
authors.affiliation: 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.
authors.forename: Yuqi
authors.initials: Y
authors.lastname: Cai
authors.affiliation: 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.
authors.forename: Jamie A
authors.initials: JA
authors.lastname: Havrilak
authors.affiliation: 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.
authors.forename: Craig S
authors.initials: CS
authors.lastname: Bolte
authors.affiliation: 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.
authors.forename: John M
authors.initials: JM
authors.lastname: Shannon
authors.affiliation: 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.
authors.forename: Tanya V
authors.initials: TV
authors.lastname: Kalin
authors.affiliation: 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. Vladimir.Kalinichenko@cchmc.org.
authors.forename: Vladimir V
authors.initials: VV
authors.lastname: Kalinichenko
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000235:genetics; D001483:Base Sequence; D001808:Blood Vessels / Q000196:embryology / Q000378:metabolism*; D015153:Blotting, Western; D002460:Cell Line; D002478:Cells, Cultured; D004622:Embryo, Mammalian / Q000166:cytology / Q000196:embryology / Q000378:metabolism; D042783:Endothelial Cells / Q000378:metabolism*; D051858:Forkhead Transcription Factors / Q000235:genetics / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental; D006801:Humans; D007150:Immunohistochemistry; D008168:Lung / Q000098:blood supply / Q000196:embryology / Q000378:metabolism; D008648:Mesoderm / Q000166:cytology / Q000378:metabolism; D051379:Mice; D057507:Mice, 129 Strain; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012689:Sequence Homology, Nucleic Acid; D015398:Signal Transduction; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism*
chemical_list: D051858:Forkhead Transcription Factors; C486293:Foxf1 protein, mouse; D042461:Vascular Endothelial Growth Factor A
keywords: developmental biology; endothelial cells; pulmonary circulation; vascular endothelial growth factor A
references.citation: Dev Cell. 2009 Feb;16(2):180-95
references.pmid: 19217421
references.citation: Case Rep Genet. 2012;2012:490408
references.pmid: 23074687
references.citation: Genome Res. 2013 Jan;23(1):23-33
references.pmid: 23034409
references.citation: Mol Cell Biol. 2013 Jan;33(2):371-86
references.pmid: 23149934
references.citation: Mol Cell Biol. 2007 Apr;27(7):2486-98
references.pmid: 17261592
references.citation: Mol Cell Biol. 2010 Nov;30(22):5381-93
references.pmid: 20837707
references.citation: Development. 2001 Jan;128(2):155-66
references.pmid: 11124112
references.citation: Nature. 1995 Jul 6;376(6535):62-6
references.pmid: 7596435
references.citation: Am J Physiol Lung Cell Mol Physiol. 2004 Mar;286(3):L521-30
references.pmid: 14607778
references.citation: Dev Biol. 2009 Dec 15;336(2):266-79
references.pmid: 19835856
references.citation: Mol Cell Biol. 2012 Oct;32(19):3838-50
references.pmid: 22826436
references.citation: Dev Cell. 2013 Jan 28;24(2):206-14
references.pmid: 23369715
references.citation: Cell. 1996 Dec 27;87(7):1171-80
references.pmid: 8980224
references.citation: Genesis. 2011 Apr;49(4):342-59
references.pmid: 21305688
references.citation: Dev Biol. 2010 Nov 15;347(2):301-14
references.pmid: 20816795
references.citation: Am J Respir Crit Care Med. 2011 Jul 15;184(2):172-9
references.pmid: 21471096
references.citation: Am J Physiol Lung Cell Mol Physiol. 2002 Jun;282(6):L1253-65
references.pmid: 12003781
references.citation: Nature. 1996 Apr 4;380(6573):439-42
references.pmid: 8602242
references.citation: J Biol Chem. 2009 Feb 27;284(9):5936-44
references.pmid: 19049965
references.citation: J Clin Invest. 2012 Nov;122(11):3862-72
references.pmid: 23023706
references.citation: J Biol Chem. 2002 Apr 5;277(14):12369-74
references.pmid: 11809759
references.citation: Blood. 2012 Sep 27;120(13):2562-72
references.pmid: 22855605
references.citation: Mech Dev. 1997 Dec;69(1-2):53-69
references.pmid: 9486531
references.citation: Am J Physiol Lung Cell Mol Physiol. 2001 Apr;280(4):L695-704
references.pmid: 11238010
references.citation: Development. 2001 Jun;128(12):2397-406
references.pmid: 11493558
references.citation: Hepatology. 2003 Jan;37(1):107-17
references.pmid: 12500195
references.citation: Am J Hum Genet. 2009 Jun;84(6):780-91
references.pmid: 19500772
references.citation: Am J Respir Cell Mol Biol. 2008 Oct;39(4):390-9
references.pmid: 18421012
references.citation: Nature. 1996 Apr 4;380(6573):435-9
references.pmid: 8602241
references.citation: Proc Natl Acad Sci U S A. 2008 Dec 9;105(49):19330-5
references.pmid: 19033457
references.citation: Cell. 2008 Dec 12;135(6):1053-64
references.pmid: 19070576
references.citation: Dev Biol. 2001 Jul 15;235(2):489-506
references.pmid: 11437453
references.citation: Nature. 1995 Jul 6;376(6535):66-70
references.pmid: 7596436
references.citation: Genesis. 2008 Feb;46(2):74-80
references.pmid: 18257043