FIGURE 7.

Epigenetic modifiers increase γδ T cell treatment efficacy. (A) Dose–response curves of bicellular A375‐fibroblast spheroids, A2058‐fibroblast spheroids, patient derived melanoma spheroids and melanoma patient‐derived organoids (MPDOs) after treatment with Entinostat, Decitabine, JQ‐1, Tubastatin A, Valproic acid, Ricolinostat, Vorinostat and DMSO controls for 3 days. Values represent mean ± SEM, n = 4. (B) Expression of MICA/B, FasL, PD‐L1 and PD‐L2 in cells from A2058‐fibroblast spheroids was measured by FACS after Entinostat, Decitabine, JQ‐1, Tubastatin A, Valproic acid, Ricolinostat and Vorinostat treatment (5 μM) for 48 h. *p < 0.05. **p < 0.01. (C) Western blot analysis of HDAC6, HDAC7, COX2, phospho‐STAT3, total STAT3, phosphor‐ERK, total ERK, phospho‐JNK, total JNK and GAPDH proteins in A2058‐fibroblast spheroids after Entinostat, Decitabine, JQ‐1, Tubastatin A, Valproic acid, Ricolinostat and Vorinostat treatment (5 μM) for 48 h (left panel). Densitometry quantification of the HDAC7, HDAC6 and COX2 (right panel). **p < 0.01. (D) NKG2A, PD‐1, NKG2D, CXCR3. CD107a and intracellular IFN‐γ in γδ T cells was measured by FACS after Entinostat, Decitabine, JQ‐1, Tubastatin A, Valproic acid, Ricolinostat and Vorinostat treatment (5 μM) for 48 h. *p < 0.05. **p < 0.01. (E) Cytotoxic function of γδ T cells against A2058‐fibro spheroids and MPDOs combined with 1 μM of Entinostat, Decitabine, JQ‐1, Tubastatin A, Valproic acid, Ricolinostat, Vorinostat and DMSO controls. n = 4. *p < 0.05. **p < 0.01