Fig. 4: LLC0150 is an NSD1/2 PROTAC with preferential cytotoxicity in AR-driven PCa.
From: NSD2 is a requisite subunit of the AR/FOXA1 neo-enhanceosome in promoting prostate tumorigenesis
a, Structure of LLC0150 and schema of NSD1 and NSD2 functional domains. LLC0150-binding PWWP1 ___domain is highlighted using a dashed red box. HMG: High mobility group; PHD: Plant homeodomain. b, Immunoblots of listed proteins in LNCaP cells treated with UNC6934 (warhead), LLC0150-dead (epimer control) or LLC0150 for 12 h at 1 μM. Total histone H3 is used as a loading control. c, Immunoblots of listed proteins in VCaP cells treated with LLC0150 (2uM) for increasing time durations. Total histone H3 is used as a loading control. d, GSEA plots of MYC target genes using the fold-change rank-ordered genes from LLC0150 vs DMSO treated LNCaP cells. DEGS, differentially expressed genes (n = 2 biological replicates; GSEA enrichment test). e, Venn diagram showing the overlap of AR ChIP-seq peaks in LNCaP cells treated with LLC0150 (2 μM for 48 h) or DMSO as control. f, ChIP-seq read-density heatmaps of AR, FOXA1, and H3K27ac at enhancers that are co-bound by AR and FOXA1 in LNCaP cells plus/minus treatment with LLC0150 (2 μM for 48 h). g, Percent growth inhibition (Cell-titer Glo) of LNCaP cells upon co-treatment with varying concentrations of LLC0150 and enzalutamide. h, Dose-response curves of LLC0150 or enzalutamide in parental or enzalutamide-resistant VCaP cells. Data are presented as mean ± SEM (n = 2 biological replicates). Serving as a control, enzalutamide dose-response curve credentials the enzalutamide-resistant VCaP cell line. i, IC50 rank-order plot of over 110 human-derived normal or cancer cell lines after 5 days of treatment with LLC0150. AR+ PCa models are highlighted in red, and NSD2-mutant hematologic cell lines are shown in purple as well as marked with an asterisk (*). Each cell line’s originating tissue lineages and known NSD2 alteration status are shown below.
