Fig. 7: NKX2-1 and FOXA2 recruit p300/CBP to activate NE enhancers and induce NEPC tumor growth, which can be abolished by p300/CBP inhibition.
a, Volcano plot showing FOXA2-interacting proteins in LuNE cells by MS (pooled data from three Co-IP replicates). The x axis represents log2(FC), and the y axis represents −log10P by two-sided t test. Each dot represents a protein. b, Co-IP showing that FOXA2 and NKX2-1 interact with CBP and p300 in LuNE cells. Data shown are from one of three (n = 3) independent experiments. c, Co-IP showing that NKX2-1 interacts with FOXA2 and p300 in NCI-H660 cells. Data are shown as in b (n = 3). d, Heatmaps showing indicated ChIP–seq intensity centered (±2 kb) around the five clusters of FOXA2-binding sites identified in the time-course LNCaP+FOXA2 cells. Scale bar—enrichment intensity. e, Heatmap showing that genes induced by FOXA2 and NKX2-1 (FC ≥ 2 and adjusted P < 0.05) are regulated by p300/CBP in LuNE cells. Data shown is the log2(FC) of sgNKX2-1, sgFOXA2 or sgFOXA2+sgNKX2-1 relative to sgNC, as well as of shp300, shCBP or shp300+shCBP to shCtrl. Adjusted P values by Wald test with Benjamini–Hochberg correction. f, LuNE cells were treated with DMSO or CCS1477 (250 nM) for 72 h. Differentially expressed genes were identified by DESeq2 with FC ≥ 2, adjusted P < 0.05. Color bar—z score. Adjusted P values are calculated as in e. g, GO analysis of CCS1477-repressed genes identified in f. Top enriched molecular concepts are shown on the y axis, while the x axis indicates enrichment. P values by one-sided hypergeometric test. h, Gene set enrichment analysis (GSEA) showing enrichment of SE-associated genes in LuNE cells treated with DMSO or CCS1477. P value by two-sided permutation test with Benjamini–Hochberg correction. i,j. Tumor growth curve (i) and weight (j) of LuNE xenograft tumors treated with vehicle or CCS1477 (n = 6 mice per group). Data are mean ± s.e.m, and P values by two-sided t test. k,l. Tumor growth curve (k) and weight (l) of LuCaP145.2 xenograft tumors treated with vehicle or CCS1477 (n = 6 mice per group in k, n = 7 tumors per group in l). Data are mean ± s.d, and P values by two-sided t test. m, Representative IHC images (n = 6 mice per group) of H3K27ac, SOX2, NKX2-1 and Ki-67 in LuCaP145.2 xenograft tumors treated with vehicle or CCS1477. Scale bar = 30 µm. Insets are shown at higher magnification (×40).
