Biased GLP-2 agonist with strong G protein-coupling but impaired arrestin recruitment and receptor desensitization enhances intestinal growth in mice

Maria Buur Nordskov Gabe, Liv von Voss Christensen, Jenna Elizabeth Hunt, Sarina Gadgaard, Lærke Smidt Gasbjerg, Jens Juul Holst, Hannelouise Kissow, Bolette Hartmann, Mette Marie Rosenkilde*

*Corresponding author af dette arbejde

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Abstract

BACKGROUND AND PURPOSE: Glucagon-like peptide-2 (GLP-2) is secreted postprandially by enteroendocrine L-cells and stimulates growth of the gut and bone. One GLP-2 analogue is approved for short bowel syndrome (SBS). To improve therapeutic efficacy, we developed biased GLP-2 receptor (GLP-2R) agonists through N-terminal modifications.

EXPERIMENTAL APPROACH: Variants with Ala and Trp substitutions of the first seven positions of GLP-2(1-33) were studied in vitro for affinity, G protein activation (cAMP accumulation), recruitment of β-arrestin 1 and 2, and internalization of the human and mouse GLP-2R. The intestinotrophic actions of the most efficacious (cAMP) biased variant was examined in mice.

KEY RESULTS: Overall, Ala substitutions had more profound effects than Trp substitutions. For both, alterations at positions 1, 3 and 6 most severely impaired activity. β-arrestin recruitment was more affected than cAMP accumulation. Among the Ala substitutions, [H1A], [D3A] and [F6A] impaired potency for cAMP-accumulation >20-fold and efficacy (E max ) to 48-87%, and were unable to recruit arrestins. The Trp substitutions, [A2W], [D3W] and [G4W] were also partial agonists (E max of 46-59%) with 1.7-12-fold decreased potencies in cAMP and diminished β-arrestin recruitment. The biased variants, [F6A], [F6W] and [S7W] induced less GLP-2R internalization compared to GLP-2, which induced internalization in a partly arrestin-independent manner. In mice, [S7W] enhanced the gut trophic actions with increased weight of the small intestine, increased villus height and crypt depth compared to GLP-2.

CONCLUSION AND IMPLICATIONS: G protein-biased GLP-2R agonists with diminished receptor desensitization have superior intestinotrophic effect and could represent improved treatment of intestinal insufficiency including SBS.

OriginalsprogEngelsk
TidsskriftBritish Journal of Pharmacology
Vol/bind180
Udgave nummer13
Sider (fra-til)1674-1689
ISSN0007-1188
DOI
StatusUdgivet - jan. 2023

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