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. 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）激活所诱导的表观基因组应答，我们采用了小鼠前列腺组织的三维类器官模型。以10 nM二氢睾酮（dihydrotestosterone, DHT）或溶剂对照处理对照（正常野生型）与SPOP突变型前列腺类器官。随后，我们开展了转录组分析（mRNA测序，mRNA-seq），同时进行染色质开放状态图谱分析（ATAC测序，ATAC-seq），并检测了转录因子（AR、FOXA1）的结合特征以及H3K4me2修饰核小体的分布情况。我们对正常（野生型）及SPOP突变型小鼠前列腺类器官中的H3K4me2组蛋白修饰、AR与FOXA1结合特征进行了检测分析。针对H3K4me2与FOXA1的染色质免疫共沉淀测序（ChIP-seq）实验，我们分别对经DHT与乙醇（EtOH）处理的细胞完成了该实验；而针对AR的ChIP-seq实验，仅对经DHT处理的细胞进行。