SPOP mutation reshapes chromatin landscape and transcriptional response to androgens [ChIP-seq]

To define the epigenomic response to AR activation, we employed the 3D organoid model of murine prostate tissue. Control and SPOP-mutant prostate organoids were stimulated with 10 nM dihydrotestosterone (DHT) or vehicle. Next, we performed the transcriptomic analysis (mRNA-seq) together with profiling of the accessibility landscape (ATAC-seq), transcription factor (AR, and FOXA1) binding and H3K4me2 modified nucleosomes. Overall design: Examination of H3K4me2 histone modifications, and AR and FOXA1 binding in Control (normal) and SPOP-mutant mouse prostate organoids. In case of H3K4me2 and FOXA1 ChIP-seq cells we performed ChIP-seq on DHT and EtOH treated cells . In case of AR ChIP-eq, only on DHT-treated cells.

为明确雄激素受体（AR）激活引发的表观基因组应答，我们采用了小鼠前列腺组织的3D类器官模型。将野生型对照与SPOP突变型前列腺类器官分别以10纳摩尔（nM）二氢睾酮（DHT）及溶剂对照进行刺激处理。随后，我们同步开展了转录组分析（mRNA测序，mRNA-seq）、染色质开放区域图谱分析（ATAC测序，ATAC-seq）、转录因子（AR与FOXA1）结合特征检测，以及H3K4me2修饰核小体的分布谱分析。整体实验设计：检测野生型（正常）与SPOP突变型小鼠前列腺类器官中的H3K4me2组蛋白修饰水平、AR与FOXA1的结合情况。针对H3K4me2与FOXA1的染色质免疫沉淀测序（ChIP测序，ChIP-seq），我们分别对DHT处理组与乙醇（EtOH）处理组细胞开展了测序实验；而针对AR的染色质免疫沉淀测序（ChIP-seq），则仅对DHT处理组细胞进行检测。