Fig. 5: Sulfated oxysterols promote monocyte-DC differentiation and antitumor activity.

A Mice bearing established LLC tumors were treated with the sulfated oxysterols 25-HC-3S, PFM037 or vehicle. At sacrifice, absolute numbers of Ly6C^highCD11c⁺MHC-II⁺ monocyte-DCs/mg of LLC tumors, were evaluated. Mean and s.d. of 4 mice/group. *P < 0.05 (Anova). B LLC-bearing mice were treated with the sulfated oxysterols 25-HC-3S, PFM037 or vehicle and monitored for tumor growth. Mean and s.d. of one experiment (n = 6 mice/group). *P < 0.05; ****P < 0.0001 (Anova). C Mice-bearing established RMA tumors were treated with the sulfated oxysterol 25-HC-3S, PFM037 or vehicle. At sacrifice, absolute numbers of Ly6C^highCD11c⁺MHC-II⁺ monocyte-DCs/mg of RMA tumors, were evaluated. Mean and s.d. of 7-8 mice/group. *P < 0.05 (Anova). D RMA-bearing mice were treated with the sulfated oxysterols 25-HC-3S, PFM037 or vehicle and evaluated for tumor weight at mice sacrifice. Mean and s.d. of 7-8 mice/group. **P < 0.01; ***P < 0.001 (Anova). E-H Tumor weights (E) and numbers of mono-DCs (F), cDC2 identified by the expression of CD26 and CD64 markers (G), and Inf-DC2 identified by the expression of CD26, CD64 and MAR-1 markers (H) infiltraing tumors from mice treated with vehicle or PFM037. Mean and s.d. of 6-7 mice/group. ns, not significant; *P < 0.05; **P < 0.01 (Student’s t-test). The difference of the numbers of mono-DCs between the experiments reported in A and F is due to the different days of analysis after tumor challenge and treatment, 12 and 10 days in A and F, respectively. I Melanoma B16F1-OVA growth delay following the treatment with PFM037. B16F1-OVA-bearing mice were treated with vehicle or with PFM037. Mean and s.d. of one experiment (n = 8 mice/group). **P < 0.01; ***P < 0.001; ****P < 0.0001 (Student’s t-test). (J) Representative images of liver metastases 21 days after injection of CRC in mice treated with vehicle (gray frame) or PFM037 (black frame). The black dashed line indicates tumor lesions. (K) Representative H&E and GFP immunohistochemical micrographs showing liver tumor features of metastases found in vehicle- (gray frame) or PFM037-treated (black frame) mice 21 days after intramesenteric injection of MC38-GFP cells. Scale bar = 300μm. L, M Quantification of the number of macroscopic liver tumor lesions (L), as well as the percentage of tumor area (M) determined by IHC in the mice described in J-K. Mean values ± s.d. are shown; *P < 0.05 (Mann-Whitney test). N Plasma levels of total cholesterol, triglycerides (TG), high and low-density lipoproteins (HDL, LDL), and high-density lipoproteins (HDL), Not Esterified Fatty Acids, and in vehicle and ratio between total cholesterol and HDL in vehicle and PFM037 (0.4 μg/day)-treated mice. *P < 0.05; ***P < 0.001; ****P < 0.0001 (Anova).