Figure 4.

Combination trametinib and sapanisertib impeded tumor metastases in a MM metastatic model. A metastatic subline of M1 was injected hematogenously into nude mice via tail vein. Four treatment groups included vehicle control, daily (V); sapanisertib, 3 times a week (S), trametinib, daily (T), and combination (T+S); n=10 mice per group. Treatments started approximately one week after tumor cell injection and extended four weeks. Trametinib was given at 5.0 mg/Kg and sapanisertib 2.0 mg/Kg. The intra-thoracic tumor burden was evaluated using H&E stained sections of thoracic tissues using quantitative machine learning pattern classifier image analyses. A) Representative illustrations of H&E-stained tissue photomicrographs of a vehicle control and a combination treated mouse, and the corresponding morphometric pseudocolor markup of the identical images segmented as tumor (red) and lung plus other non-neoplastic thoracic tissues (green) are shown (bars=1 mm). B) Ratios of tumor to total intra-thoracic tissue, the latter representing tumor plus non-neoplastic tissue areas, by treatment group. Five step sections of thoracic tissues, approximately 400 μm apart, were evaluated for each animal (sections per treatment = 45 – 50). Each point represents analysis of individual tissue sections. Comparisons of the tumor burdens between groups displayed as box plots depicting median counts, solid line, with 1^st and 3^rd quartile ranges indicated. Wilcoxon signed-rank tests were used to compare the experimental metastatic areas between treatment groups. p-values are indicated.