Yap1 overexpression and its targeting in Kmt2d- and Trp53-mutated B-cell lymphoma. (A) YAP1 mRNA expression measured by real-time quantitative polymerase chain reaction in 6 human DLBCL cell lines (n = 3 for each) with and without KMT2D and/or TP53 mutations. (B) YAP1 and pYAP1 protein expression measured by capillary electrophoresis-based immunoassay in 6 human DLBCL cell lines. Lamin B1 is used as a loading control. Representative of 3 independent experiments. (C) Effect of CA3 treatment on viability of 6 human DLBCL cell lines (n = 3 for each) measured by CellTiter-Glo luminescent cell viability assay. (D) Cell proliferation of Toledo cells expressing short hairpin RNA (shRNA) targeting GFP (control) or YAP1 after puromycin selection. (E) YAP1 protein expression measured by capillary electrophoresis-based immunoassay in Toledo cells expressing shRNA targeting GFP or YAP1. Lamin B1 is used as a loading control. Representative of 2 independent experiments. (F) Tumor volume in Toledo xenograft mice administered with 4 mg/kg of CA3 (n = 7) or vehicle (n = 7) 3 times a week for 12 doses. (G) Survival curves of mice transplanted with 2.5 × 103 murine double sgKmt2d-targeted/Trp53-altered B-cell lymphoma cells and administered with 4 mg/kg of CA3 (n = 16) or vehicle (n = 14) 3 times a week for 6 doses. Log-rank test. In panels C-D,F, data are represented as mean ± standard deviation. In panels A,D,F, Welch t test was performed. ∗P < .05; ∗∗P < .005; ∗∗∗P < .0005.
