GLP-1 (7–36) and glucagon dose dependently stimulated cAMP production and calcitonin secretion. Exendin (9–39) abolished a further increase in cAMP formation at glucagon concentration over 10 nM and partially suppressed glucagon-induced calcitonin secretion.
cAMP production, calcitonin mRNA levels, and calcitonin secretion
GLP-1 (7–37) increased cAMP formation in a dose-dependent manner. Exedin (9–39), an antagonist of GLP-1 receptor, blunted the stimulation of cAMP production induced by GLP-1 (7–37). Gene expression and peptide secretion of calcitonin were increased after incubation of CA-77 cells with GLP-1 (7–37).
Human TT C-cell line, rat MTC 6–23 C-cell line, and rat CA-77 C-cell line
GLP-1 receptor mRNA and protein expression; calcitonin release after GLP-1 receptor agonists
Native GLP-1, liraglutide, and exenatide all stimulate calcitonin gene expression and calcitonin secretion via the GLP-1 receptor in a dose-dependent manner in rat C cells. The human TT cells express few GLP-1 receptors compared with rat MTC 6–23 and CA-77 and show a lack of functional response to GLP-1 and GLP-1 receptor agonists.
Sprague Dawley rats aged 6-7 weeks and CD-1 mice at the age of 5–10 weeks
Plasma calcitonin and pathological examination to thyroid gland sections after dosing with liraglutide
Calcitonin levels increase with time and dose with 104-week repeated dosing of liraglutide. The incidences of both C-cell hyperplasia and C-cell tumor formation at 104 weeks were increased in a dose-dependent manner and reached statistical significance.
CD-1 wild-type mice aged 5-6 weeks and GLP-1-receptor knockout mice at the age of 4-5 weeks
Plasma calcitonin, pathological examination to thyroid tissue sections, and immunohistochemical staining for phosphoproteins after 13-week treatment with liraglutide or exenatide
GLP-1 agonists cause calcitonin release and C-cell hyperplasia in wild-type mice via a GLP-1-receptor-dependent mechanism. GLP-1 activates the mammalian target of rapamycin (mTOR) pathway by stimulating the production of cAMP. Activation of mTOR in turn results in downstream phosphorylation of ribosome S6. Liraglutide-induced C-cell hyperplasia in mice is not associated with RET activation.
Plasma calcitonin and pathological analysis to thyroid gland sections after dosing with liraglutide
No increase in plasma calcitonin was seen in cynomolgus monkeys receiving a single dose of liraglutide or during 87-week daily dose. There was also no change in the thyroid gland sections, relative C-cell fraction of the thyroid gland, and proliferative index in the C cells after liraglutide for 52 weeks.
Geometric mean levels of serum calcitonin and outlier analysis
There was no significant difference in mean calcitonin levels between liraglutide and control groups. The proportions of subjects with calcitonin levels shifting to a higher category or above a clinically relevant cut-off value of 20 pg/mL were low and did not differ between treatment groups.