title: Temporal and spatial regulation of epsin abundance and VEGFR3 signaling are required for lymphatic valve formation and function.
abstract: Lymphatic valves prevent the backflow of the lymph fluid and ensure proper lymphatic drainage throughout the body. Local accumulation of lymphatic fluid in tissues, a condition called lymphedema, is common in individuals with malformed lymphatic valves. The vascular endothelial growth factor receptor 3 (VEGFR3) is required for the development of lymphatic vascular system. The abundance of VEGFR3 in collecting lymphatic trunks is high before valve formation and, except at valve regions, decreases after valve formation. We found that in mesenteric lymphatics, the abundance of epsin 1 and 2, which are ubiquitin-binding adaptor proteins involved in endocytosis, was low at early stages of development. After lymphatic valve formation, the initiation of steady shear flow was associated with an increase in the abundance of epsin 1 and 2 in collecting lymphatic trunks, but not in valve regions. Epsin 1 and 2 bound to VEGFR3 and mediated the internalization and degradation of VEGFR3, resulting in termination of VEGFR3 signaling. Mice with lymphatic endothelial cell-specific deficiency of epsin 1 and 2 had dilated lymphatic capillaries, abnormally high VEGFR3 abundance in collecting lymphatics, immature lymphatic valves, and defective lymph drainage. Deletion of a single Vegfr3 allele or pharmacological suppression of VEGFR3 signaling restored normal lymphatic valve development and lymph drainage in epsin-deficient mice. Our findings establish a critical role for epsins in the temporal and spatial regulation of VEGFR3 abundance and signaling in collecting lymphatic trunks during lymphatic valve formation.
authors.affiliation: Cardiovascular Biology Program, Oklahoma Medical Research Foundation, Oklahoma, OK 73104, USA. Department of Biochemistry and Molecular Biology, University of Oklahoma Health Science Center, Oklahoma, OK 73104, USA.
authors.forename: Xiaolei
authors.initials: X
authors.lastname: Liu
authors.affiliation: Cardiovascular Biology Program, Oklahoma Medical Research Foundation, Oklahoma, OK 73104, USA.
authors.forename: Satish
authors.initials: S
authors.lastname: Pasula
authors.affiliation: Cardiovascular Biology Program, Oklahoma Medical Research Foundation, Oklahoma, OK 73104, USA.
authors.forename: Hoogeun
authors.initials: H
authors.lastname: Song
authors.affiliation: Cardiovascular Biology Program, Oklahoma Medical Research Foundation, Oklahoma, OK 73104, USA.
authors.forename: Kandice L
authors.initials: KL
authors.lastname: Tessneer
authors.affiliation: Cardiovascular Biology Program, Oklahoma Medical Research Foundation, Oklahoma, OK 73104, USA.
authors.forename: Yunzhou
authors.initials: Y
authors.lastname: Dong
authors.affiliation: Cardiovascular Biology Program, Oklahoma Medical Research Foundation, Oklahoma, OK 73104, USA.
authors.forename: Scott
authors.initials: S
authors.lastname: Hahn
authors.affiliation: Cardiovascular Biology Program, Oklahoma Medical Research Foundation, Oklahoma, OK 73104, USA.
authors.forename: Tadayuki
authors.initials: T
authors.lastname: Yago
authors.affiliation: Cardiovascular Biology Program, Oklahoma Medical Research Foundation, Oklahoma, OK 73104, USA. Department of Biochemistry and Molecular Biology, University of Oklahoma Health Science Center, Oklahoma, OK 73104, USA.
authors.forename: Megan L
authors.initials: ML
authors.lastname: Brophy
authors.affiliation: Cardiovascular Biology Program, Oklahoma Medical Research Foundation, Oklahoma, OK 73104, USA.
authors.forename: Baojun
authors.initials: B
authors.lastname: Chang
authors.affiliation: Cardiovascular Biology Program, Oklahoma Medical Research Foundation, Oklahoma, OK 73104, USA.
authors.forename: Xiaofeng
authors.initials: X
authors.lastname: Cai
authors.affiliation: Cardiovascular Biology Program, Oklahoma Medical Research Foundation, Oklahoma, OK 73104, USA.
authors.forename: Hao
authors.initials: H
authors.lastname: Wu
authors.affiliation: Cardiovascular Biology Program, Oklahoma Medical Research Foundation, Oklahoma, OK 73104, USA.
authors.forename: John
authors.initials: J
authors.lastname: McManus
authors.affiliation: Laboratory of Developmental Genetics, Center for Experimental Medicine and Systems Biology, Institute of Medical Science, University of Tokyo, Minato-ku, Tokyo 108-8639, Japan.
authors.forename: Hirotake
authors.initials: H
authors.lastname: Ichise
authors.affiliation: Arthritis and Clinical Immunology Research Program, Oklahoma Medical Research Foundation, Oklahoma, OK 73104, USA.
authors.forename: Constantin
authors.initials: C
authors.lastname: Georgescu
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Oklahoma Health Science Center, Oklahoma, OK 73104, USA. Arthritis and Clinical Immunology Research Program, Oklahoma Medical Research Foundation, Oklahoma, OK 73104, USA.
authors.forename: Jonathan D
authors.initials: JD
authors.lastname: Wren
authors.affiliation: Cardiovascular Biology Program, Oklahoma Medical Research Foundation, Oklahoma, OK 73104, USA. Department of Cell Biology, University of Oklahoma Health Sciences Center, Oklahoma, OK 73126, USA.
authors.forename: Courtney
authors.initials: C
authors.lastname: Griffin
authors.affiliation: Cardiovascular Biology Program, Oklahoma Medical Research Foundation, Oklahoma, OK 73104, USA. Department of Biochemistry and Molecular Biology, University of Oklahoma Health Science Center, Oklahoma, OK 73104, USA.
authors.forename: Lijun
authors.initials: L
authors.lastname: Xia
authors.affiliation: Cardiovascular Biology Program, Oklahoma Medical Research Foundation, Oklahoma, OK 73104, USA.
authors.forename: R Sathish
authors.initials: RS
authors.lastname: Srinivasan
authors.affiliation: Cardiovascular Biology Program, Oklahoma Medical Research Foundation, Oklahoma, OK 73104, USA. Department of Biochemistry and Molecular Biology, University of Oklahoma Health Science Center, Oklahoma, OK 73104, USA. hong-chen@omrf.org.
authors.forename: Hong
authors.initials: H
authors.lastname: Chen
mesh_terms: D033942:Adaptor Proteins, Vesicular Transport / Q000378:metabolism*; D000483:Alleles; D000818:Animals; D049109:Cell Proliferation; D002469:Cell Separation; D003433:Crosses, Genetic; D004705:Endocytosis; D042783:Endothelial Cells / Q000166:cytology; D005260:Female; D005434:Flow Cytometry; D049452:Green Fluorescent Proteins / Q000737:chemistry; D007211:Indoles / Q000737:chemistry; D008024:Ligands; D008198:Lymph Nodes / Q000473:pathology; D008208:Lymphatic System / Q000378:metabolism*; D008209:Lymphedema / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009281:Naphthalenes / Q000737:chemistry; D010957:Plasmids / Q000378:metabolism; D015398:Signal Transduction; D013997:Time Factors; D040321:Vascular Endothelial Growth Factor Receptor-3 / Q000378:metabolism*
chemical_list: C441602:3-(4-dimethylamino-naphthalen-1-ylmethylene)-1,3-dihydro-indol-2-one; D033942:Adaptor Proteins, Vesicular Transport; C577774:Epn2 protein, mouse; D007211:Indoles; D008024:Ligands; D009281:Naphthalenes; C114154:epsin; D049452:Green Fluorescent Proteins; D040321:Vascular Endothelial Growth Factor Receptor-3