Fig. 3. Semaglutide-induced weight loss fails to improve tumor immunity.

Mice were placed on either normal chow (NC) or western diet (WD) for 12 weeks. A At 12 weeks, a cohort of mice on WD began twice weekly treatment with semaglutide (WD+Sema) for 6 weeks while being maintained on WD. B Body mass was tracked over time, and graphed data is from a representative experiment showing the average mass of all mice in each group with SEM indicated by the error bars. C After 6 weeks, fat mass and lean mass were assessed via whole-body NMR imaging (NC n = 10, WD n = 10, WD+Sema n = 20), D liver histology was performed, E and serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), and cholesterol levels were measured (NC n = 10, WD n = 10, WD+Sema n = 30). At this time, mice received a subcutaneous injection of B16 tumor cells in the flank (day 0) and half of each cohort was treated with immune checkpoint blockade (ICB; anti-PD-1/anti-CTLA-4) or received PBS vehicle control on days 6 and 10. F Tumor volumes were measured every other day starting at day 6 and G pooled tumor volumes from 2 independent experiments at day 16 were compared (NC n = 18, NC + ICB n = 7, WD n = 19, WD + ICB n = 14, WD+Sema n = 35, WD+Sema+ICB n = 35). H Tumors were harvested at day 16 and representative FACS plots show intracellular expression of GzmB and Perforin by TRP-2-specific CD8⁺ TIL. I Pooled data from 2 independent experiments show the percent of TRP-2-specific CD8⁺ TIL co-expressing GzmB and Perforin (NC n = 9, NC + ICB n = 3, WD n = 9, WD + ICB n = 7, WD+Sema n = 17, WD+Sema+ICB n = 19). All box-and-whisker plots: The box indicates the 25^th and 75^th percentile, the line indicates the data median, and the whiskers indicate the minimum and maximum of all individual values. All n’s represent an individual mouse. Exact P values were calculated by two-sided Mann–Whitney U test.