title: FKBPL is a critical antiangiogenic regulator of developmental and pathological angiogenesis.
abstract: OBJECTIVE
  The antitumor effects of FK506-binding protein like (FKBPL) and its extracellular role in angiogenesis are well characterized; however, its role in physiological/developmental angiogenesis and the effect of FKBPL ablation has not been evaluated. This is important as effects of some angiogenic proteins are dosage dependent. Here we evaluate the regulation of FKBPL secretion under angiogenic stimuli, as well as the effect of FKBPL ablation in angiogenesis using mouse and zebrafish models.
  
  
  RESULTS
  FKBPL is secreted maximally by human microvascular endothelial cells and fibroblasts, and this was specifically downregulated by proangiogenic hypoxic signals, but not by the angiogenic cytokines, VEGF or IL8. FKBPL's critical role in angiogenesis was supported by our inability to generate an Fkbpl knockout mouse, with embryonic lethality occurring before E8.5. However, whilst Fkbpl heterozygotic embryos showed some vasculature irregularities, the mice developed normally. In murine angiogenesis models, including the ex vivo aortic ring assay, in vivo sponge assay, and tumor growth assay, Fkbpl(+/-) mice exhibited increased sprouting, enhanced vessel recruitment, and faster tumor growth, respectively, supporting the antiangiogenic function of FKBPL. In zebrafish, knockdown of zFkbpl using morpholinos disrupted the vasculature, and the phenotype was rescued with hFKBPL. Interestingly, this vessel disruption was ineffective when zcd44 was knocked-down, supporting the dependency of zFkbpl on zCd44 in zebrafish.
  
  
  CONCLUSIONS
  FKBPL is an important regulator of angiogenesis, having an essential role in murine and zebrafish blood vessel development. Mouse models of angiogenesis demonstrated a proangiogenic phenotype in Fkbpl heterozygotes.
authors.affiliation: From the McClay Research Centre for Pharmaceutical Sciences, School of Pharmacy (A.Y., R.B., M.H., O.L., A.S., H.M., L.N., C.M.M., R.M., L.M., H.O.M., T.R.), Centre for Infection and Immunity (M.H., O.L., A.K.), and Northern Ireland Molecular Pathology Laboratory, Centre for Cancer Research and Cell Biology (K.A.), School of Medicine, Dentistry and Biomedical Sciences, Queen's University, Belfast, UK; School of Immunity and Infection and Cancer Studies, Institute for Biomedical Research, University of Birmingham, Birmingham, UK (I.H.-N., R.B.); Centre for Digestive Diseases, Queen Mary, University of London, Barts and The London School of Medicine and Dentistry, London, UK (J.-M.D.); and Birth Defects Research Centre, UCL Institute of Child Health, London, UK (J.-M.D., A.J.B.).
authors.forename: Anita
authors.initials: A
authors.lastname: Yakkundi
authors.affiliation: From the McClay Research Centre for Pharmaceutical Sciences, School of Pharmacy (A.Y., R.B., M.H., O.L., A.S., H.M., L.N., C.M.M., R.M., L.M., H.O.M., T.R.), Centre for Infection and Immunity (M.H., O.L., A.K.), and Northern Ireland Molecular Pathology Laboratory, Centre for Cancer Research and Cell Biology (K.A.), School of Medicine, Dentistry and Biomedical Sciences, Queen's University, Belfast, UK; School of Immunity and Infection and Cancer Studies, Institute for Biomedical Research, University of Birmingham, Birmingham, UK (I.H.-N., R.B.); Centre for Digestive Diseases, Queen Mary, University of London, Barts and The London School of Medicine and Dentistry, London, UK (J.-M.D.); and Birth Defects Research Centre, UCL Institute of Child Health, London, UK (J.-M.D., A.J.B.).
authors.forename: Rachel
authors.initials: R
authors.lastname: Bennett
authors.affiliation: From the McClay Research Centre for Pharmaceutical Sciences, School of Pharmacy (A.Y., R.B., M.H., O.L., A.S., H.M., L.N., C.M.M., R.M., L.M., H.O.M., T.R.), Centre for Infection and Immunity (M.H., O.L., A.K.), and Northern Ireland Molecular Pathology Laboratory, Centre for Cancer Research and Cell Biology (K.A.), School of Medicine, Dentistry and Biomedical Sciences, Queen's University, Belfast, UK; School of Immunity and Infection and Cancer Studies, Institute for Biomedical Research, University of Birmingham, Birmingham, UK (I.H.-N., R.B.); Centre for Digestive Diseases, Queen Mary, University of London, Barts and The London School of Medicine and Dentistry, London, UK (J.-M.D.); and Birth Defects Research Centre, UCL Institute of Child Health, London, UK (J.-M.D., A.J.B.).
authors.forename: Ivette
authors.initials: I
authors.lastname: Hernández-Negrete
authors.affiliation: From the McClay Research Centre for Pharmaceutical Sciences, School of Pharmacy (A.Y., R.B., M.H., O.L., A.S., H.M., L.N., C.M.M., R.M., L.M., H.O.M., T.R.), Centre for Infection and Immunity (M.H., O.L., A.K.), and Northern Ireland Molecular Pathology Laboratory, Centre for Cancer Research and Cell Biology (K.A.), School of Medicine, Dentistry and Biomedical Sciences, Queen's University, Belfast, UK; School of Immunity and Infection and Cancer Studies, Institute for Biomedical Research, University of Birmingham, Birmingham, UK (I.H.-N., R.B.); Centre for Digestive Diseases, Queen Mary, University of London, Barts and The London School of Medicine and Dentistry, London, UK (J.-M.D.); and Birth Defects Research Centre, UCL Institute of Child Health, London, UK (J.-M.D., A.J.B.).
authors.forename: Jean-Marie
authors.initials: JM
authors.lastname: Delalande
authors.affiliation: From the McClay Research Centre for Pharmaceutical Sciences, School of Pharmacy (A.Y., R.B., M.H., O.L., A.S., H.M., L.N., C.M.M., R.M., L.M., H.O.M., T.R.), Centre for Infection and Immunity (M.H., O.L., A.K.), and Northern Ireland Molecular Pathology Laboratory, Centre for Cancer Research and Cell Biology (K.A.), School of Medicine, Dentistry and Biomedical Sciences, Queen's University, Belfast, UK; School of Immunity and Infection and Cancer Studies, Institute for Biomedical Research, University of Birmingham, Birmingham, UK (I.H.-N., R.B.); Centre for Digestive Diseases, Queen Mary, University of London, Barts and The London School of Medicine and Dentistry, London, UK (J.-M.D.); and Birth Defects Research Centre, UCL Institute of Child Health, London, UK (J.-M.D., A.J.B.).
authors.forename: Mary
authors.initials: M
authors.lastname: Hanna
authors.affiliation: From the McClay Research Centre for Pharmaceutical Sciences, School of Pharmacy (A.Y., R.B., M.H., O.L., A.S., H.M., L.N., C.M.M., R.M., L.M., H.O.M., T.R.), Centre for Infection and Immunity (M.H., O.L., A.K.), and Northern Ireland Molecular Pathology Laboratory, Centre for Cancer Research and Cell Biology (K.A.), School of Medicine, Dentistry and Biomedical Sciences, Queen's University, Belfast, UK; School of Immunity and Infection and Cancer Studies, Institute for Biomedical Research, University of Birmingham, Birmingham, UK (I.H.-N., R.B.); Centre for Digestive Diseases, Queen Mary, University of London, Barts and The London School of Medicine and Dentistry, London, UK (J.-M.D.); and Birth Defects Research Centre, UCL Institute of Child Health, London, UK (J.-M.D., A.J.B.).
authors.forename: Oksana
authors.initials: O
authors.lastname: Lyubomska
authors.affiliation: From the McClay Research Centre for Pharmaceutical Sciences, School of Pharmacy (A.Y., R.B., M.H., O.L., A.S., H.M., L.N., C.M.M., R.M., L.M., H.O.M., T.R.), Centre for Infection and Immunity (M.H., O.L., A.K.), and Northern Ireland Molecular Pathology Laboratory, Centre for Cancer Research and Cell Biology (K.A.), School of Medicine, Dentistry and Biomedical Sciences, Queen's University, Belfast, UK; School of Immunity and Infection and Cancer Studies, Institute for Biomedical Research, University of Birmingham, Birmingham, UK (I.H.-N., R.B.); Centre for Digestive Diseases, Queen Mary, University of London, Barts and The London School of Medicine and Dentistry, London, UK (J.-M.D.); and Birth Defects Research Centre, UCL Institute of Child Health, London, UK (J.-M.D., A.J.B.).
authors.forename: Kenneth
authors.initials: K
authors.lastname: Arthur
authors.affiliation: From the McClay Research Centre for Pharmaceutical Sciences, School of Pharmacy (A.Y., R.B., M.H., O.L., A.S., H.M., L.N., C.M.M., R.M., L.M., H.O.M., T.R.), Centre for Infection and Immunity (M.H., O.L., A.K.), and Northern Ireland Molecular Pathology Laboratory, Centre for Cancer Research and Cell Biology (K.A.), School of Medicine, Dentistry and Biomedical Sciences, Queen's University, Belfast, UK; School of Immunity and Infection and Cancer Studies, Institute for Biomedical Research, University of Birmingham, Birmingham, UK (I.H.-N., R.B.); Centre for Digestive Diseases, Queen Mary, University of London, Barts and The London School of Medicine and Dentistry, London, UK (J.-M.D.); and Birth Defects Research Centre, UCL Institute of Child Health, London, UK (J.-M.D., A.J.B.).
authors.forename: Amy
authors.initials: A
authors.lastname: Short
authors.affiliation: From the McClay Research Centre for Pharmaceutical Sciences, School of Pharmacy (A.Y., R.B., M.H., O.L., A.S., H.M., L.N., C.M.M., R.M., L.M., H.O.M., T.R.), Centre for Infection and Immunity (M.H., O.L., A.K.), and Northern Ireland Molecular Pathology Laboratory, Centre for Cancer Research and Cell Biology (K.A.), School of Medicine, Dentistry and Biomedical Sciences, Queen's University, Belfast, UK; School of Immunity and Infection and Cancer Studies, Institute for Biomedical Research, University of Birmingham, Birmingham, UK (I.H.-N., R.B.); Centre for Digestive Diseases, Queen Mary, University of London, Barts and The London School of Medicine and Dentistry, London, UK (J.-M.D.); and Birth Defects Research Centre, UCL Institute of Child Health, London, UK (J.-M.D., A.J.B.).
authors.forename: Hayley
authors.initials: H
authors.lastname: McKeen
authors.affiliation: From the McClay Research Centre for Pharmaceutical Sciences, School of Pharmacy (A.Y., R.B., M.H., O.L., A.S., H.M., L.N., C.M.M., R.M., L.M., H.O.M., T.R.), Centre for Infection and Immunity (M.H., O.L., A.K.), and Northern Ireland Molecular Pathology Laboratory, Centre for Cancer Research and Cell Biology (K.A.), School of Medicine, Dentistry and Biomedical Sciences, Queen's University, Belfast, UK; School of Immunity and Infection and Cancer Studies, Institute for Biomedical Research, University of Birmingham, Birmingham, UK (I.H.-N., R.B.); Centre for Digestive Diseases, Queen Mary, University of London, Barts and The London School of Medicine and Dentistry, London, UK (J.-M.D.); and Birth Defects Research Centre, UCL Institute of Child Health, London, UK (J.-M.D., A.J.B.).
authors.forename: Laura
authors.initials: L
authors.lastname: Nelson
authors.affiliation: From the McClay Research Centre for Pharmaceutical Sciences, School of Pharmacy (A.Y., R.B., M.H., O.L., A.S., H.M., L.N., C.M.M., R.M., L.M., H.O.M., T.R.), Centre for Infection and Immunity (M.H., O.L., A.K.), and Northern Ireland Molecular Pathology Laboratory, Centre for Cancer Research and Cell Biology (K.A.), School of Medicine, Dentistry and Biomedical Sciences, Queen's University, Belfast, UK; School of Immunity and Infection and Cancer Studies, Institute for Biomedical Research, University of Birmingham, Birmingham, UK (I.H.-N., R.B.); Centre for Digestive Diseases, Queen Mary, University of London, Barts and The London School of Medicine and Dentistry, London, UK (J.-M.D.); and Birth Defects Research Centre, UCL Institute of Child Health, London, UK (J.-M.D., A.J.B.).
authors.forename: Cian M
authors.initials: CM
authors.lastname: McCrudden
authors.affiliation: From the McClay Research Centre for Pharmaceutical Sciences, School of Pharmacy (A.Y., R.B., M.H., O.L., A.S., H.M., L.N., C.M.M., R.M., L.M., H.O.M., T.R.), Centre for Infection and Immunity (M.H., O.L., A.K.), and Northern Ireland Molecular Pathology Laboratory, Centre for Cancer Research and Cell Biology (K.A.), School of Medicine, Dentistry and Biomedical Sciences, Queen's University, Belfast, UK; School of Immunity and Infection and Cancer Studies, Institute for Biomedical Research, University of Birmingham, Birmingham, UK (I.H.-N., R.B.); Centre for Digestive Diseases, Queen Mary, University of London, Barts and The London School of Medicine and Dentistry, London, UK (J.-M.D.); and Birth Defects Research Centre, UCL Institute of Child Health, London, UK (J.-M.D., A.J.B.).
authors.forename: Ross
authors.initials: R
authors.lastname: McNally
authors.affiliation: From the McClay Research Centre for Pharmaceutical Sciences, School of Pharmacy (A.Y., R.B., M.H., O.L., A.S., H.M., L.N., C.M.M., R.M., L.M., H.O.M., T.R.), Centre for Infection and Immunity (M.H., O.L., A.K.), and Northern Ireland Molecular Pathology Laboratory, Centre for Cancer Research and Cell Biology (K.A.), School of Medicine, Dentistry and Biomedical Sciences, Queen's University, Belfast, UK; School of Immunity and Infection and Cancer Studies, Institute for Biomedical Research, University of Birmingham, Birmingham, UK (I.H.-N., R.B.); Centre for Digestive Diseases, Queen Mary, University of London, Barts and The London School of Medicine and Dentistry, London, UK (J.-M.D.); and Birth Defects Research Centre, UCL Institute of Child Health, London, UK (J.-M.D., A.J.B.).
authors.forename: Lana
authors.initials: L
authors.lastname: McClements
authors.affiliation: From the McClay Research Centre for Pharmaceutical Sciences, School of Pharmacy (A.Y., R.B., M.H., O.L., A.S., H.M., L.N., C.M.M., R.M., L.M., H.O.M., T.R.), Centre for Infection and Immunity (M.H., O.L., A.K.), and Northern Ireland Molecular Pathology Laboratory, Centre for Cancer Research and Cell Biology (K.A.), School of Medicine, Dentistry and Biomedical Sciences, Queen's University, Belfast, UK; School of Immunity and Infection and Cancer Studies, Institute for Biomedical Research, University of Birmingham, Birmingham, UK (I.H.-N., R.B.); Centre for Digestive Diseases, Queen Mary, University of London, Barts and The London School of Medicine and Dentistry, London, UK (J.-M.D.); and Birth Defects Research Centre, UCL Institute of Child Health, London, UK (J.-M.D., A.J.B.).
authors.forename: Helen O
authors.initials: HO
authors.lastname: McCarthy
authors.affiliation: From the McClay Research Centre for Pharmaceutical Sciences, School of Pharmacy (A.Y., R.B., M.H., O.L., A.S., H.M., L.N., C.M.M., R.M., L.M., H.O.M., T.R.), Centre for Infection and Immunity (M.H., O.L., A.K.), and Northern Ireland Molecular Pathology Laboratory, Centre for Cancer Research and Cell Biology (K.A.), School of Medicine, Dentistry and Biomedical Sciences, Queen's University, Belfast, UK; School of Immunity and Infection and Cancer Studies, Institute for Biomedical Research, University of Birmingham, Birmingham, UK (I.H.-N., R.B.); Centre for Digestive Diseases, Queen Mary, University of London, Barts and The London School of Medicine and Dentistry, London, UK (J.-M.D.); and Birth Defects Research Centre, UCL Institute of Child Health, London, UK (J.-M.D., A.J.B.).
authors.forename: Alan J
authors.initials: AJ
authors.lastname: Burns
authors.affiliation: From the McClay Research Centre for Pharmaceutical Sciences, School of Pharmacy (A.Y., R.B., M.H., O.L., A.S., H.M., L.N., C.M.M., R.M., L.M., H.O.M., T.R.), Centre for Infection and Immunity (M.H., O.L., A.K.), and Northern Ireland Molecular Pathology Laboratory, Centre for Cancer Research and Cell Biology (K.A.), School of Medicine, Dentistry and Biomedical Sciences, Queen's University, Belfast, UK; School of Immunity and Infection and Cancer Studies, Institute for Biomedical Research, University of Birmingham, Birmingham, UK (I.H.-N., R.B.); Centre for Digestive Diseases, Queen Mary, University of London, Barts and The London School of Medicine and Dentistry, London, UK (J.-M.D.); and Birth Defects Research Centre, UCL Institute of Child Health, London, UK (J.-M.D., A.J.B.).
authors.forename: Roy
authors.initials: R
authors.lastname: Bicknell
authors.affiliation: From the McClay Research Centre for Pharmaceutical Sciences, School of Pharmacy (A.Y., R.B., M.H., O.L., A.S., H.M., L.N., C.M.M., R.M., L.M., H.O.M., T.R.), Centre for Infection and Immunity (M.H., O.L., A.K.), and Northern Ireland Molecular Pathology Laboratory, Centre for Cancer Research and Cell Biology (K.A.), School of Medicine, Dentistry and Biomedical Sciences, Queen's University, Belfast, UK; School of Immunity and Infection and Cancer Studies, Institute for Biomedical Research, University of Birmingham, Birmingham, UK (I.H.-N., R.B.); Centre for Digestive Diseases, Queen Mary, University of London, Barts and The London School of Medicine and Dentistry, London, UK (J.-M.D.); and Birth Defects Research Centre, UCL Institute of Child Health, London, UK (J.-M.D., A.J.B.).
authors.forename: Adrien
authors.initials: A
authors.lastname: Kissenpfennig
authors.affiliation: From the McClay Research Centre for Pharmaceutical Sciences, School of Pharmacy (A.Y., R.B., M.H., O.L., A.S., H.M., L.N., C.M.M., R.M., L.M., H.O.M., T.R.), Centre for Infection and Immunity (M.H., O.L., A.K.), and Northern Ireland Molecular Pathology Laboratory, Centre for Cancer Research and Cell Biology (K.A.), School of Medicine, Dentistry and Biomedical Sciences, Queen's University, Belfast, UK; School of Immunity and Infection and Cancer Studies, Institute for Biomedical Research, University of Birmingham, Birmingham, UK (I.H.-N., R.B.); Centre for Digestive Diseases, Queen Mary, University of London, Barts and The London School of Medicine and Dentistry, London, UK (J.-M.D.); and Birth Defects Research Centre, UCL Institute of Child Health, London, UK (J.-M.D., A.J.B.). t.robson@qub.ac.uk.
authors.forename: Tracy
authors.initials: T
authors.lastname: Robson
mesh_terms: D000818:Animals; D001011:Aorta / Q000378:metabolism*; D018827:Carcinoma, Lewis Lung / Q000098:blood supply* / Q000378:metabolism* / Q000473:pathology; D015687:Cell Hypoxia; D005260:Female; D018507:Gene Expression Regulation, Developmental; D005838:Genotype; D006801:Humans; D018960:Hyaluronan Receptors / Q000235:genetics / Q000378:metabolism; D020104:Immunophilins / Q000235:genetics / Q000378:metabolism*; D061986:MCF-7 Cells; D008297:Male; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D009389:Neovascularization, Pathologic; D018919:Neovascularization, Physiologic; D010641:Phenotype; D015398:Signal Transduction; D022021:Tacrolimus Binding Proteins / Q000235:genetics / Q000378:metabolism*; D013997:Time Factors; D047368:Tumor Burden; D015027:Zebrafish; D029961:Zebrafish Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: C400101:FKBPL protein, human; D018960:Hyaluronan Receptors; D029961:Zebrafish Proteins; C000598204:FKBPL protein, mouse; C000598205:FKBPL protein, zebrafish; D022021:Tacrolimus Binding Proteins; D020104:Immunophilins
keywords: CD44; FKBPL; angiogenesis; knockout; vasculature