Journal: Neoplasia (New York, N.Y.)

Article Title: Genetic loss of CHD1 regulates distinct histone post-translational modifications in the development of castration-resistant prostate cancer

doi: 10.1016/j.neo.2026.101289

Figure Lengend Snippet: CHD1 knockout reduces histone H3K36 methylation and NSD2 robustly in CRPC cell lines. A. Immunoblots showing CHD1, NSD2, and EZH2 levels in CHD1-WT and -KO CRPC cells, DU145 and 22RV-1CR, respectively. The blots were quantified using ImageJ and are shown as a percentage of the control CHD1-WT condition. B. The stacked bar plots depict the percent abundance of H3.3K36 (left) and H3.3K27 (right) methylation levels in the CRPC DU145 cells, based on CHD1 status. Each level of methylation is colored/patterned according to the legend shown at the top of the graph. C. H3K36me2 expression at the protein level in human CRPC TMA samples ( n = 48). The samples were stratified into CHD1-high (above median, n = 23) and CHD1-low (below median, n = 25) groups based on CHD1 staining quantification. Representative IHC staining images of H3K36me2 in CHD1-high and CHD1-low groups. The color scheme below the images depicts the staining intensity. Lowest staining (purple), highest staining (red), and unanalyzed stromal tissue (green). D. The quantification of IHC staining for H3K36me2 in CHD1-high and CHD1-low groups. The staining from only epithelial cell nuclei was analyzed ( p < 0.01). For all graphs, data is shown as a mean ± SEM.

Article Snippet: DU145, a CRPC cell line purchased from ATCC (Manassas, VA, USA), was routinely cultured in DMEM medium (Corning; Corning, NY, USA) containing 4.5 gm/L glucose supplemented with 10 % fetal bovine serum (FBS) (Gibco, Waltham, MA, USA) and 100 units/mL penicillin/streptomycin (PS) (Corning).

Techniques: Knock-Out, Methylation, Western Blot, Control, Expressing, Staining, Immunohistochemistry

Journal: Neoplasia (New York, N.Y.)

Article Title: Genetic loss of CHD1 regulates distinct histone post-translational modifications in the development of castration-resistant prostate cancer

doi: 10.1016/j.neo.2026.101289

Figure Lengend Snippet: CHD1 status alters H3K4me3 and H3K27me3 levels at NSD2 promoter and regulates its transcription. A. ChIP-based qPCR for binding of CHD1, H3K4me3, and H3K27me3 was assessed on the promoter region of the NSD2 gene and B. EZH2 gene in CHD1-WT and -KO DU145 cells. The schema shows the location of different sites assessed at the promoter, transcription start site, and exonic regions ( p < 0.05). C. ChIP-based qPCR for binding of CHD1 and H3K4me3 for NSD2 promoter region (sites shown in A) in CHD1-WT and -KO 22RV1-CR cells and D. LNCaP-CR cells ( p < 0.05). E. ChIP-based qPCR for binding of CHD1 and H3K4me3 for EZH2 promoter region (sites shown in B) in CHD1-WT and -KO 22RV1-CR cells and F. LNCaP-CR cells ( p < 0.05). G. The relative mRNA expression of NSD2 and H. EZH2 in CHD1-WT and -KO CRPC DU145 and 22RV-1CR cells with RT-qPCR ( p < 0.05). For all graphs, data is shown as a mean ± SEM.

Article Snippet: DU145, a CRPC cell line purchased from ATCC (Manassas, VA, USA), was routinely cultured in DMEM medium (Corning; Corning, NY, USA) containing 4.5 gm/L glucose supplemented with 10 % fetal bovine serum (FBS) (Gibco, Waltham, MA, USA) and 100 units/mL penicillin/streptomycin (PS) (Corning).

Techniques: ChIP-qPCR, Binding Assay, Expressing, Quantitative RT-PCR

Journal: Neoplasia (New York, N.Y.)

Article Title: Genetic loss of CHD1 regulates distinct histone post-translational modifications in the development of castration-resistant prostate cancer

doi: 10.1016/j.neo.2026.101289

Figure Lengend Snippet: Transcriptomic profiling and ChIP analysis identify interferon signaling and its target genes to be downregulated with CHD1 knockout. A. Volcano plot of differentially expressed genes with CHD1-KO in DU145 cells. CHD1-KO results in downregulation of 4382 genes (FDR<0.01, log 2 FC<−0.2). B. Gene set enrichment analysis (GSEA) using Hallmark gene set identifies interferon gamma and alpha signaling as top pathways de-enriched with CHD1-KO in DU145 cells (FDR<0.05, p < 0.05). C. ChIP-qPCR for binding of H3K36me2 on the 5′, gene body, and 3′ regions of different viral mimicry and IFN downstream targets identified from GSEA core enriched genes in CHD1-WT and -KO DU145 cells. The H3K36me2 binding is reduced with CHD1-KO ( p < 0.05). For all graphs, data is shown as a mean ± SEM.

Article Snippet: DU145, a CRPC cell line purchased from ATCC (Manassas, VA, USA), was routinely cultured in DMEM medium (Corning; Corning, NY, USA) containing 4.5 gm/L glucose supplemented with 10 % fetal bovine serum (FBS) (Gibco, Waltham, MA, USA) and 100 units/mL penicillin/streptomycin (PS) (Corning).

Techniques: Knock-Out, ChIP-qPCR, Binding Assay

Journal: Neoplasia (New York, N.Y.)

Article Title: Genetic loss of CHD1 regulates distinct histone post-translational modifications in the development of castration-resistant prostate cancer

doi: 10.1016/j.neo.2026.101289

Figure Lengend Snippet: Differential sensitivity of CHD1-WT and -KO CRPC cells to NSD2 inhibition in vitro . A. Cell proliferation assay with different concentrations of NSD2 PROTAC degrader UNC8153 in CHD1-WT and -KO DU145 cells, B. 22RV1-CR cells, and C. LNCaP-CR cells. UNC8153-mediated NSD2 degradation showed that CHD1-WT CRPC cells are more sensitive to NSD2 inhibition on day 7 ( p < 0.05). For all graphs, data is shown as a mean ± SEM. D. Proposed model for the evolution of castration-resistant tumors in either the CHD1 wildtype or deficient state. As androgen resistance develops, functional CHD1 is essential to maintain an open chromatin state and H3K4me3 levels at the NSD2 promoter, thereby regulating NSD2 transcription. In contrast, CHD1 deficiency leads to increased heterochromatin formation and elevated H3K27me3 levels. CHD1-intact CRPC cells are more sensitive to NSD2 inhibition than CHD1-deficient CRPC cells, suggesting that CHD1 may function as a predictive biomarker for NSD2 drug response.

Article Snippet: DU145, a CRPC cell line purchased from ATCC (Manassas, VA, USA), was routinely cultured in DMEM medium (Corning; Corning, NY, USA) containing 4.5 gm/L glucose supplemented with 10 % fetal bovine serum (FBS) (Gibco, Waltham, MA, USA) and 100 units/mL penicillin/streptomycin (PS) (Corning).

Techniques: Inhibition, In Vitro, Proliferation Assay, Functional Assay, Biomarker Discovery