PUBLICATION

The endosomal RIN2/Rab5C machinery prevents VEGFR2 degradation to control gene expression and tip cell identity during angiogenesis

Authors
Kempers, L., Wakayama, Y., van der Bijl, I., Furumaya, C., De Cuyper, I.M., Jongejan, A., Kat, M., van Stalborch, A.D., van Boxtel, A.L., Hubert, M., Geerts, D., van Buul, J.D., de Korte, D., Herzog, W., Margadant, C.
ID
ZDB-PUB-210514-9
Date
2021
Source
Angiogenesis   24(3): 695-714 (Journal)
Registered Authors
Herzog, Wiebke
Keywords
Early endosomes, Endolysosomal trafficking, Notch signaling, Rab GTPases, Rab5C, Sprouting angiogenesis, Tip cells, VEGF signaling, VEGFR2
MeSH Terms
  • Carrier Proteins/genetics
  • Carrier Proteins/metabolism*
  • Neovascularization, Physiologic*
  • Humans
  • Proteolysis*
  • Vascular Endothelial Growth Factor Receptor-2/genetics
  • Vascular Endothelial Growth Factor Receptor-2/metabolism*
  • Human Umbilical Vein Endothelial Cells/metabolism*
  • Guanine Nucleotide Exchange Factors/genetics
  • Guanine Nucleotide Exchange Factors/metabolism*
  • Zebrafish/genetics
  • Zebrafish/metabolism*
  • rab5 GTP-Binding Proteins/genetics
  • rab5 GTP-Binding Proteins/metabolism*
  • Animals
  • Gene Expression Regulation*
(all 16)
PubMed
33983539 Full text @ Angiogenesis
Abstract
Sprouting angiogenesis is key to many pathophysiological conditions, and is strongly regulated by vascular endothelial growth factor (VEGF) signaling through VEGF receptor 2 (VEGFR2). Here we report that the early endosomal GTPase Rab5C and its activator RIN2 prevent lysosomal routing and degradation of VEGF-bound, internalized VEGFR2 in human endothelial cells. Stabilization of endosomal VEGFR2 levels by RIN2/Rab5C is crucial for VEGF signaling through the ERK and PI3-K pathways, the expression of immediate VEGF target genes, as well as specification of angiogenic 'tip' and 'stalk' cell phenotypes and cell sprouting. Using overexpression of Rab mutants, knockdown and CRISPR/Cas9-mediated gene editing, and live-cell imaging in zebrafish, we further show that endosomal stabilization of VEGFR2 levels is required for developmental angiogenesis in vivo. In contrast, the premature degradation of internalized VEGFR2 disrupts VEGF signaling, gene expression, and tip cell formation and migration. Thus, an endosomal feedforward mechanism maintains receptor signaling by preventing lysosomal degradation, which is directly linked to the induction of target genes and cell fate in collectively migrating cells during morphogenesis.
Genes / Markers
Figures
Figure Gallery (7 images)
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Expression
Phenotype
Fish Conditions Stage Phenotype Figure
WT + MO1-rab5ccontrolPrim-15
mu227Tg; s843Tgstandard conditionsPrim-15
mu227Tg; s843Tgstandard conditionsPrim-15
mu227Tg; s843Tgstandard conditionsPrim-15
mu227Tg; s843Tgstandard conditionsPrim-15
mu228Tg; s843Tgstandard conditionsPrim-15
mu228Tg; s843Tgstandard conditionsPrim-15
mu228Tg; s843Tgstandard conditionsPrim-15
mu228Tg; s843Tgstandard conditionsPrim-15
mu240TgcontrolPrim-15
1 - 10 of 29
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Mutations / Transgenics
Allele Construct Type Affected Genomic Region
mu227TgTransgenic Insertion
    mu228TgTransgenic Insertion
      mu229
        		Small Deletion
        mu240TgTransgenic Insertion
          s843TgTransgenic Insertion
            s940TgTransgenic Insertion
              1 - 6 of 6
              Show
              Human Disease / Model
              No data available
              Sequence Targeting Reagents
              Target Reagent Reagent Type
              rab5cCRISPR1-rab5cCRISPR
              rab5cCRISPR2-rab5cCRISPR
              rab5cMO1-rab5cMRPHLNO
              rin2aMO1-rin2aMRPHLNO
              1 - 4 of 4
              Show
              Fish
              Fish
              mu227Tg; s843Tg
              mu227Tg; s843Tg + MO1-rin2a
              mu228Tg; s843Tg
              mu240Tg
              mu240Tg + MO1-rab5c
              rab5cmu229/mu229; s843Tg
              s843Tg + MO1-rab5c
              s843Tg + MO1-rin2a
              s843Tg; s940Tg
              s843Tg; s940Tg + MO1-rab5c
              1 - 10 of 12
              Show
              Antibodies
              Orthology
              No data available
              Engineered Foreign Genes
              Marker Marker Type Name
              EGFPEFGEGFP
              mCherryEFGmCherry
              VenusEFGVenus
              1 - 3 of 3
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              Mapping
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              Citation
              Kempers, L., Wakayama, Y., van der Bijl, I., Furumaya, C., De Cuyper, I.M., Jongejan, A., Kat, M., van Stalborch, A.D., van Boxtel, A.L., Hubert, M., Geerts, D., van Buul, J.D., de Korte, D., Herzog, W., Margadant, C. (2021) The endosomal RIN2/Rab5C machinery prevents VEGFR2 degradation to control gene expression and tip cell identity during angiogenesis. Angiogenesis. 24(3):695-714.