Fig. 7.

In vitro and in vivo effects of mouse HK2 ASOs, as single agents and in combination with DPI, MET and PER. (A) mHK2-ASO1 suppresses HK2 expression in HK1⁻HK2⁺ P3 cells. (B) The combinations of mHK2-ASO1/DPI/PER and mHK2-ASO1/MET/PER induce apoptosis in HK1⁻HK2⁺ P3 cells. Cells were pre-treated with mHK2-ASO1 (0.5 μM) for 3 days, followed by the addition of MET (2 mM), DPI (25 nM), and PER (3 μM) for another 3 days. Apoptosis (APO) was determined by quantification of the Annexin V-positive population. (C) 8-day treatments with mHK2-ASO1, mHK2-ASO1/MET/PER, and mHK2-ASO1/DPI/PER significantly reduced ¹⁸F-FDG PET signal in mouse HK1⁻HK2⁺ P3 tumors. Left, PET scan images are shown. Right, quantification of PET signal intensity from tumor ROI values. (D) Effects of mHK2-ASO1, mHK2-ASO1/MET/PER, and mHK2-ASO1/DPI/PER on HK1⁻HK2⁺ P3 tumor progression. When tumors reached 200 mm³ (day 1), mice were randomized into indicated groups (n = 11/group). mHK2-ASO1 treatment started on day 1 at 50 mg/kg s.c. daily. DPI (2 mg/kg, daily i.p.), MET (250 mg/kg, daily i.p.), and PER (30 mg/kg, daily i.p.) started on day 3. Treatments were given for 5 days per week, with a 2-day break between weeks. NS, not significant. (E) Body weight of the four indicated groups during treatments. (F) Disease progression in the P3/HK1⁻/mCherry-LUC disseminated model was monitored by BLI. BLI measurement in photons per second per cm² per steradian (p/s/cm²/sr) was translated to color to indicate disease activity in the mice by the scales shown at far right. (G) Quantification of whole mouse BLI photons in panel F. (H) Kaplan-Meier survival curves of mice in panel F. *, P < 0.05. **, P < 0.01. ***, P < 0.001. ****, P < 0.0001.