In vivo functional profiling and structural characterisation of the human Glp1r A316T variant.

Glucagon-like peptide-1 receptor (GLP-1R) agonists (GLP-1RAs) are a highly effective therapy class for type 2 diabetes (T2D) and obesity, yet there are variable patient responses. Variation in the human Glp1r gene leading to altered receptor structure, signal transduction, and function might be directly linked to variable therapeutic responses in patients. A naturally occurring, low-frequency, gain-of-function missense variant, rs10305492 G>A (A316T), protects against T2D and cardiovascular disease. Here we employ CRISPR/Cas9 technology to generate a humanised knock-in mouse model bearing the homozygous Glp1r A316T substitution. Human Glp1r ^A316T/A316T mice displayed lower fasting blood glucose levels and improved glucose tolerance, as well as increased plasma insulin levels and improved insulin secretion compared to human Glp1r ^+/+ littermates, even under metabolic stress. They also exhibited alterations in islet cytoarchitecture and β-cell identity under a high-fat, high-sucrose (HFHS) diet. This was however associated with blunted responses to pharmacological GLP-1RAs in vivo . Further investigations in several rodent and human β-cell models demonstrated that the human Glp1r A316T variant exhibits characteristics of constitutive activation but dampened GLP-1RA responses. Our results are further supported by the cryo-EM analysis and molecular dynamics (MD) simulations of the GLP-1R A316T structure, collectively demonstrating that the A316T Glp1r variant governs basal receptor activity and pharmacological responses to GLP-1R-targeting anti-diabetic therapies, highlighting the importance of the molecular characterisation of human Glp1r variants to predict individual therapy responses.