hGCGR/hGLP1R

Fig.1 Body weight change of HFD-induced hGCGR/hGLP1R obese mice treated with Cotadutide. In HFD-induced hGCGR/hGLP1R obese mice, Cotadutide significantly decreased body weight. The hGCGR/hGLP1R mice were fed CD or HFD. PBS or Cotadutide was administered subcutaneously once daily until euthanasia. (A) Body weight and (B) body weight change were recorded (n=6, Mean ± SEM, ***p<0.001, Group2 vs. Group3). 

Abbr: CD, chow diet. HFD, high-fat diet. 

Fig.2 Food intake change of HFD-induced hGCGR/hGLP1R obese mice treated with Cotadutide. In HFD-induced hGCGR/hGLP1R obese mice, Cotadutide significantly decreased food intake. The hGCGR/hGLP1R mice were fed CD or HFD. PBS or Cotadutide was administered subcutaneously once daily until euthanasia. Food intake per mouse was recorded (n=6, Mean ± SEM, ***p<0.001, Group2 vs. Group3). 

Abbr: CD, chow diet. HFD, high-fat diet. 

Fig.3 Fat mass and lean mass changes of HFD-induced hGCGR/hGLP1R obese mice treated with Cotadutide. In HFD-induced hGCGR/hGLP1R obese mice, Cotadutide significantly decreased both fat mass and lean mass; however, the lean mass-to-body weight ratio increased compared with HFD-fed control. The hGCGR/hGLP1R mice were fed CD or HFD. PBS or Cotadutide was administered subcutaneously once daily until euthanasia. At the study endpoint, fat and lean mass were determined. The absolute masses (A, B) were recorded, the weight changes (C, D) were calculated, and the fat mass-to-body weight and lean mass-to-body weight ratios (E, F) were computed for each compartment (n=6, Mean ± SEM, *p<0.05,  ***p<0.001). 

Abbr: CD, chow diet. HFD, high-fat diet. 

Fig.4 Liver triglyceride and cholesterol content of HFD-induced hGCGR/hGLP1R obese mice treated with Cotadutide. In HFD-induced hGCGR/hGLP1R obese mice, Cotadutide significantly decreased liver total cholesterol. There was no significant difference in liver triglycerides content among the groups. The hGCGR/hGLP1R mice were fed CD or HFD. PBS or Cotadutide was administered subcutaneously once daily until euthanasia. Liver tissues were homogenized, and liver (A) triglycerides (TG) and (B) total cholesterol (T-CHO) levels were determined (n=6, Mean ± SEM, *p<0.05, **p<0.01). 

Abbr: CD, chow diet. HFD, high-fat diet. 

Fig.5 Blood glucose levels of HFD-induced hGCGR/hGLP1R obese mice treated with Cotadutide. In HFD-induced hGCGR/hGLP1R obese mice, Cotadutide significantly decreased blood glucose and improved glucose tolerance. The hGCGR/hGLP1R mice were fed CD or HFD. PBS or Cotadutide was administered subcutaneously once daily until euthanasia. (A) An intraperitoneal glucose tolerance test (IPGTT) was performed by intraperitoneally injecting glucose and measuring blood glucose levels at 0, 15, 30, 60, 90 and 120 min post-injection (***p<0.001, Group2 vs. Group3). Subsequently, (B) the area under the curve (AUC) was computed to evaluate systemic glucose tolerance (n=6, Mean ± SEM, ***p<0.001). 

Abbr: CD, chow diet. HFD, high-fat diet. 

Fig.6 Serum insulin levels of HFD-induced hGCGR/hGLP1R obese mice treated with Cotadutide. In HFD-induced hGCGR/hGLP1R obese mice, Cotadutide significantly decreased serum insulin level. The hGCGR/hGLP1R mice were fed CD or HFD. PBS or Cotadutide was administered subcutaneously once daily until euthanasia. Serum insulin levels were determined at the end of the study (n=6, Mean ± SEM, *p<0.05, **p<0.01). 

Abbr: CD, chow diet. HFD, high-fat diet. 

Fig.7 White adipose tissue weight of HFD-induced hGCGR/hGLP1R obese mice treated with Cotadutide. Cotadutide reduced white adipose tissue mass, particularly in epididymal (eWAT), inguinal (iWAT) and mesenteric (mWAT) depots in HFD-induced hGCGR/hGLP1R obese mice. The hGCGR/hGLP1R mice were fed CD or HFD. PBS or Cotadutide was administered subcutaneously once daily until euthanasia. (A) Epididymal (eWAT), (B) inguinal (iWAT), and (C) mesenteric (mWAT) white adipose tissues were dissected, weighed and normalized to body weight (D-F) (n=6, Mean ± SEM, **p<0.01, ***p<0.001). 

Abbr: CD, chow diet. HFD, high-fat diet. 

Fig.8 Brown adipose tissue weight of HFD-induced hGCGR/hGLP1R obese mice treated with Cotadutide. Cotadutide reduced brown adipose tissue (BAT) weight in HFD-induced hGCGR/hGLP1R obese mice. In hGCGR/hGLP1R mice, the BAT-to-body weight ratio was comparable between the PBS and Cotadutide groups under the HFD regimen. The hGCGR/hGLP1R mice were fed CD or HFD. PBS or Cotadutide was administered subcutaneously once daily until euthanasia. (A) Brown adipose tissue (BAT) was dissected, weighed, and normalized to body weight (BW) (n=6, Mean ± SEM, *p<0.05, ***p<0.001).

Abbr: CD, chow diet. HFD, high-fat diet. 

Fig.9 Muscle weights of HFD-induced hGCGR/hGLP1R obese mice treated with Cotadutide. In HFD-induced hGCGR/hGLP1R obese mice, Cotadutide significantly increased the tibialis anterior and gastrocnemius weight-to-body weight ratios relative to the PBS-treated HFD group. The hGCGR/hGLP1R mice were fed CD or HFD. PBS or Cotadutide was administered subcutaneously once daily until euthanasia. Soleus, tibialis anterior and gastrocnemius muscles (A-C) were dissected and weighed, and each muscle weight was normalized to body weight (BW) (D-F) (n=6, Mean ± SEM, *p<0.05, **p<0.01, ***p<0.001). 

Abbr: CD, chow diet. HFD, high-fat diet. 

Fig.10 Oxygen consumption of HFD-induced hGCGR/hGLP1R obese mice treated with Cotadutide. In HFD-induced hGCGR/hGLP1R obese mice, Cotadutide significantly increased  O2 consumption. The hGCGR/hGLP1R mice were fed CD or HFD. PBS or Cotadutide was administered subcutaneously once daily until euthanasia. Mice were housed in individual metabolic cages. (A) Oxygen consumption was recorded every hour and displayed as a curve over the experimental period (***p<0.001, Group2 vs. Group3). The area under the curve (AUC) for (B) oxygen consumption was subsequently computed (n=6, Mean ± SEM, ***p<0.001). 

Abbr: CD, chow diet. HFD, high-fat diet. 

Fig.11 Carbon dioxide production of HFD-induced hGCGR/hGLP1R obese mice treated with Cotadutide. In HFD-induced hGCGR/hGLP1R obese mice, Cotadutide significantly increased carbon dioxide production. The hGCGR/hGLP1R mice were fed CD or HFD. PBS or Cotadutide was administered subcutaneously once daily until euthanasia. Mice were housed in individual metabolic cages. (A) Carbon dioxide production was recorded every hour and displayed as a curve over the experimental period (***p<0.001, Group2 vs. Group3). The area under the curve (AUC) for (B) carbon dioxide production was subsequently computed (n=6, Mean ± SEM, ***p<0.001). 

Abbr: CD, chow diet. HFD, high-fat diet. 

Fig.12  Respiratory exchange ratio of HFD-induced hGCGR/hGLP1R obese mice treated with Cotadutide. In HFD-induced hGCGR/hGLP1R obese mice, Cotadutide significantly increased respiratory exchange ratio. The hGCGR/hGLP1R mice were fed CD or HFD. PBS or Cotadutide was administered subcutaneously once daily until euthanasia. Mice were housed in individual metabolic cages. (A) Respiratory exchange ratio (RER) (***p<0.001, Group2 vs. Group3) and (B) the area under the curve (AUC) were calculated (n=6, Mean ± SEM, ***p<0.001). 

Abbr: CD, chow diet. HFD, high-fat diet. 

Fig.13 Heat production of HFD-induced hGCGR/hGLP1R obese mice treated with Cotadutide. In HFD-induced hGCGR/hGLP1R obese mice, Cotadutide significantly increased heat production. The hGCGR/hGLP1R mice were fed CD or HFD. PBS or Cotadutide was administered subcutaneously once daily until euthanasia. Mice were housed in individual metabolic cages. (A) Heat production (***p<0.001, Group2 vs. Group3) and (B) the area under the curve (AUC) were computed (n=6, Mean ± SEM, ***p<0.001). 

Abbr: CD, chow diet. HFD, high-fat diet.