TY - JOUR

T1 - Biased GLP-2 agonist with strong G protein-coupling but impaired arrestin recruitment and receptor desensitization enhances intestinal growth in mice

AU - Gabe, Maria Buur Nordskov

AU - von Voss Christensen, Liv

AU - Hunt, Jenna Elizabeth

AU - Gadgaard, Sarina

AU - Gasbjerg, Lærke Smidt

AU - Holst, Jens Juul

AU - Kissow, Hannelouise

AU - Hartmann, Bolette

AU - Rosenkilde, Mette Marie

N1 - This article is protected by copyright. All rights reserved.

PY - 2023/1

Y1 - 2023/1

N2 - 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.

AB - 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.

U2 - 10.1111/bph.16040

DO - 10.1111/bph.16040

M3 - Journal article

C2 - 36683195

VL - 180

SP - 1674

EP - 1689

JO - British Journal of Pharmacology

JF - British Journal of Pharmacology

SN - 0007-1188

IS - 13

ER -