Figure 4

Combining AKT inhibitors and bacterial therapeutics for NSCLC therapy. (a) Schematic of the fourth part of the pipeline to validate the efficacy of one of the combination therapies. (b) AKT signaling in H460 cells upon θ toxin and MK2206 treatment: S. typhimurium was grown in presence of AHL to produce θ toxin and were lysed and filtered. H460 cells in monolayer were treated with θ toxin filtrate for 30 min, with or without 10 mM MK2206. Indirect immuno-fluorescence was carried out with antibodies against total AKT (red), phospho-AKT (Ser473) (green). Nuclei were stained with DAPI (blue). (c) Response of H460 subcutaneous tumors in NSG mice to 6 treatment cohorts (n = 3 or 4 per cohort): Vehicle, Vehicle + S. typhimurium producing GFP abbr. “St”, MK2206 targeting AKT, Vehicle + S. typhimurium producing both GFP and θ toxin abbr. “Stθ”, MK2206 + St, and, MK2206 + Stθ. Treatment began once tumors reached ~ 150 cu.mm. twice a week. Live Salmonella was injected intratumorally suspended in 40 μL of sterile 1 × DPBS (vehicle) with concentration of 4.5 X 10⁷ CFU/mL followed by subcutaneous injection of AHL, a day after, to induce θ toxin secretion within the tumor. MK2206 drug was delivered by oral gavage as 240 mg/kg dose with 30% Captisol in 1 × DPBS as vehicle. Significant change in tumor size (*** = p < 0.001) was determined by two-way repeated ANOVA and error bars represent standard error. (d) Body weight of mice measured throughout the treatment schedule, error bars represent standard error (n = 3 or 4), n.s. = not significant.