Elevated expression of the adhesion GPCR ADGRL4/ELTD1 promotes endothelial sprouting angiogenesis without activating canonical GPCR signalling

ADGRL4/ELTD1 is an orphan adhesion GPCR (aGPCR) expressed in endothelial cells that regulates tumour angiogenesis. The majority of aGPCRs are orphan receptors. The Stachel Hypothesis proposes a mechanism for aGPCR activation, in which aGPCRs contain a tethered agonist (termed Stachel ) C-terminal to...

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Published in:Scientific reports Vol. 11; no. 1; pp. 8870 - 13
Main Authors: Favara, David M., Liebscher, Ines, Jazayeri, Ali, Nambiar, Madhulika, Sheldon, Helen, Banham, Alison H., Harris, Adrian L.
Format: Journal Article
Language:English
Published: London Nature Publishing Group UK 23.04.2021
Nature Publishing Group
Nature Portfolio
Subjects:
1-Phosphatidylinositol 3-kinase
631/80
631/80/86
Adhesion
AKT protein
Angiogenesis
Codon
Endothelial cells
Endothelium, Vascular - cytology
Endothelium, Vascular - metabolism
Fluorescence resonance energy transfer
G protein-coupled receptors
HEK293 Cells
Human Umbilical Vein Endothelial Cells
Humanities and Social Sciences
Humans
Hypotheses
Hypoxia-inducible factor 1a
MAP kinase
multidisciplinary
Neovascularization, Pathologic
Orphan receptors
Peptides
Phenotypes
Proteolysis
Receptors, G-Protein-Coupled - metabolism
Science
Science (multidisciplinary)
Signal Transduction
Stat3 protein
Transcription
Tumors
Up-Regulation
β-Catenin
ISSN:2045-2322, 2045-2322
Online Access:Get full text
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Summary:ADGRL4/ELTD1 is an orphan adhesion GPCR (aGPCR) expressed in endothelial cells that regulates tumour angiogenesis. The majority of aGPCRs are orphan receptors. The Stachel Hypothesis proposes a mechanism for aGPCR activation, in which aGPCRs contain a tethered agonist (termed Stachel ) C-terminal to the GPCR-proteolytic site (GPS) cleavage point which, when exposed, initiates canonical GPCR signalling. This has been shown in a growing number of aGPCRs. We tested this hypothesis on ADGRL4/ELTD1 by designing full length (FL) and C-terminal fragment (CTF) ADGRL4/ELTD1 constructs, and a range of potential Stachel peptides. Constructs were transfected into HEK293T cells and HTRF FRET, luciferase-reporter and Alphascreen GPCR signalling assays were performed. A stable ADGRL4/ELTD1 overexpressing HUVEC line was additionally generated and angiogenesis assays, signalling assays and transcriptional profiling were performed. ADGRL4/ELTD1 has the lowest GC content in the aGPCR family and codon optimisation significantly increased its expression. FL and CTF ADGRL4/ELTD1 constructs, as well as Stachel peptides, did not activate canonical GPCR signalling. Furthermore, stable overexpression of ADGRL4/ELTD1 in HUVECs induced sprouting angiogenesis, lowered in vitro anastomoses, and decreased proliferation, without activating canonical GPCR signalling or MAPK/ERK, PI3K/AKT, JNK, JAK/HIF-1α, beta catenin or STAT3 pathways. Overexpression upregulated ANTXR1 , SLC39A6 , HBB , CHRNA , ELMOD1 , JAG1 and downregulated DLL4 , KIT , CCL15 , CYP26B1 . ADGRL4/ELTD1 specifically regulates the endothelial tip-cell phenotype through yet undefined signalling pathways.
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ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-021-85408-x