Androgen receptor (AR)-mediated transcription is the primary driver of prostate cancer growth and proliferation [1, 2]. Androgen treatment induces AR binding to the DNA primarily at regulatory enhancer elements, resulting in rapid changes to transcriptional profiles [3-6]. However, the precise relationship between androgen-induced gene expression changes and AR-enhancer binding has not been characterized in the context of 3D chromatin interactions and therefore a high-resolution view of the temporal dynamics of androgen-induced gene activation is still missing.
To determine the molecular order of events associated with transcription of androgen-induced genes, we performed Hi-C, Promoter Capture Hi-C (PCHi-C), RNA-Seq and AR ChIP-Seq experiments in DHT treated prostate cancer cell lines at five time-points (control, 30min, 2hrs, 4hrs and 16hrs) and integrated the temporal data sets. We found that while androgen-induced gene activation and AR binding occurred mostly at later time-points (4hrs and 16hrs post-DHT), the majority of differential promoter-enhancer interactions were transiently present at the earliest time-point of 30mins post-DHT. At 16hrs more than half of these interactions were subsequently lost. Our results demonstrate that changes in AR-enhancer-promoter interactions precede gene activation and unexpectedly high levels of gene transcription were associated with loss of these interactions. Our findings are consistent with the conclusion that enhancer-promoter interactions transiently activate gene expression, thereby uncoupling the temporal dynamics of chromatin interactions from those of transcription.
This novel dataset provides a temporally resolved map of androgen-induced chromatin and transcription changes in prostate cancer, revealing potential causal relationships between 3D chromatin structure, AR binding at enhancers, and gene activation.