Journal: Molecular Oncology

Article Title: Combining VPS34 inhibitors with STING agonists enhances type I interferon signaling and anti‐tumor efficacy

doi: 10.1002/1878-0261.13619

Figure Lengend Snippet: VPS34 inhibitors synergize with the STING agonist ADU‐S100 in vitro . (A) Quantification of IFNB1 , CCL5 , and CXCL10 gene expression using qRT‐PCR and (B) secreted IFNβ, CCL5, and CXCL10 using Mesoscale Discovery (MSD) assays in A‐498 cells treated with DMSO control, 2 μ m SB02024 or SAR405 in combination with 50 μ m STING agonist ADU‐S100 for 24 h. Bars represent mean ± SEM of three independent experiments. (C) Quantification of Ifnb1 , Ccl5 , and Cxcl10 gene expression using qRT‐PCR and (D) secreted CCL5 and CXCL10 using MSD assays in B16‐F10 cells treated with DMSO control, 2 μ m SB02024 or SAR405 in combination with 5 μ m ADU‐S100 for 24 h. Bars represent mean ± SEM of three independent experiments (in (C) four independent experiments were run for SB02024/ADI100 combo). (E) Quantification of IFNB1 , CCL5 , and CXCL10 gene expression using qRT‐PCR and (F) secreted IFNβ, CCL5, and CXCL10 using ELISA assays in THP‐1 cells treated with DMSO control, 2 μ m SB02024 or SAR405 in combination with 50 μ m ADU‐S100 for 24 h. Bars represent mean ± SEM of three independent experiments. (G) Quantification of Ifnb1 , Ccl5 , and Cxcl10 gene expression using qRT‐PCR and (H) secreted IFNβ, CCL5, and CXCL10 using ELISA assays in DC2.4 cells treated with DMSO control, 2 μ m SB02024 or SAR405 in combination with 10 μ m ADU‐S100 for 24 h. Bars represent mean ± SEM of three independent experiments. (I) THP‐1 reporter cells (STING present or knockout) were treated with DMSO, 2 μ m of VPS34 inhibitor (SB02024 or SAR405) in combination with 5 μg·mL −1 ADU‐S100 for 24 h. Interferon‐stimulated response element (ISRE) or NFκB signal was read out via luciferase or secreted embryonic alkaline phosphatase (SEAP), respectively. Bars represent mean ± SEM of four independent experiments. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001 using one‐way ANOVA followed by Dunnett's (black asterisks indicating comparison to DMSO‐treated control) or Šidák multiple comparison test (red asterisks indicating comparison of the combination treatment to the more effective single treatment).

Article Snippet: The VPS34 inhibitor SAR405 (#S7682; Selleckchem, Houston, TX, USA), the STING agonist ADU‐S100 (#CT‐ADUS100; ChemieTek, Indianapolis, IN, USA; or #HY‐12885; MedChemExpress, Monmouth Junction, NJ, USA), and 2′,3′‐cGAMP (cGAMP) (#TLRL‐NACGA‐1; InvivoGen) were used according to manufacturer's instructions.

Techniques: In Vitro, Expressing, Quantitative RT-PCR, Control, Enzyme-linked Immunosorbent Assay, Knock-Out, Luciferase, Comparison

Journal: Molecular Oncology

Article Title: Combining VPS34 inhibitors with STING agonists enhances type I interferon signaling and anti‐tumor efficacy

doi: 10.1002/1878-0261.13619

Figure Lengend Snippet: SB02024 potentiates the response to STING agonist in tumor‐bearing mice. (A–C) B16‐F10 cells were subcutaneously implanted in C57BL/6 mice. Mice were injected intratumorally with PBS or 10 μg STING agonist ADU‐S100 (on days 9, 11, 13, and 15) in combination with daily oral administration of vehicle or 20 mg·kg −1 SB02024 by gavage starting from day 7 to day 16 after implantation. (A) Dosing schematic and tumor growth curves of n = 15 mice per treatment group. Percent tumor growth inhibition was calculated compared to vehicle control. (B) Tumor weight of n = 10 mice per treatment group sacrificed on day 17. (C) Mice survival of n = 5 mice per treatment group. Lack of survival was defined as death or tumor size > 1000 mm 3 . Data represent mean ± SEM. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001 using one‐way ANOVA followed by Šidák multiple comparison test. Survival percentage was defined using graphpad prism , and P values were calculated using the log‐rank (Mantel‐Cox) test.

Article Snippet: The VPS34 inhibitor SAR405 (#S7682; Selleckchem, Houston, TX, USA), the STING agonist ADU‐S100 (#CT‐ADUS100; ChemieTek, Indianapolis, IN, USA; or #HY‐12885; MedChemExpress, Monmouth Junction, NJ, USA), and 2′,3′‐cGAMP (cGAMP) (#TLRL‐NACGA‐1; InvivoGen) were used according to manufacturer's instructions.

Techniques: Injection, Inhibition, Control, Comparison