Antiandrogen treatment endows ZMYDN8 dependent epigenetic reprogramming to orchestrate neuroendocrine tumor transdifferentiation [ChIP-seq]

The transdifferentiation from adenocarcinoma following androgen deprivation therapy (ADT) is thought to be the primary process leading to the development of neuroendocrine prostate cancer (NEPC). However, it remains unclear how lineage factors interact with ADT to endow the lineage transition. Through an integrated analysis of NEPC-based CRISPR-Cas9 screening and scRNA-seq tracking of tumor transitions, we unveil that antiandrogen-induced ZMYND8-dependent epigenetic programming orchestrates the transdifferentiation of NEPC. We demonstrate that the ablation of Zmynd8 restricts NEPC development in Pten/Trp53/Rb1 knockout mouse models. Conversely, ASCL1-induced upregulation of ZMYND8 shapes the neuroendocrine (NE) lineage to confer resistance to AR-targeted therapy. Mechanistically, FOXM1, a key regulator in castration-resistant prostate cancer (CRPC), stabilizes ZMYND8 binding to chromatin regions harboring H3K4me1-K14ac modification and FOXM1 targeting. Antiandrogen treatment frees the SWI/SNF chromatin remodeling complex from AR, enabling its interaction with ZMYND8/FOXM1 to upregulate key NE lineage regulators (e.g., FOXA2, SOX2, and POU3F2), thus inducing transdifferentiation. Having identified ZMYND8's link to adverse disease outcomes in CRPC patients, we develop a small molecule, ZMYND8i-34, designed to selectively inhibit its histone recognition. In pre-clinical models, ZMYND8i-34 treatment effectively blocks NE transdifferentiation and curtails CRPC development. Together, our results highlight the importance of antiandrogen treatment endowing ZMYND8-dependent epigenetic reprogramming to orchestrate lineage fate and suggest that targeting ZMYND8 may hold the potential to restrict NEPC development and overcome treatment resistance. Overall design: ChIP-seq data for LNCap cells with or without ZMYND8-OE uncovered ZMYND8 binding places change during the ADT progress on ZMYND8-OE condition.

雄激素剥夺治疗（ADT）后腺癌向神经内分泌前列腺癌（NEPC）转分化，被认为是NEPC发生的核心驱动过程。然而，谱系因子如何与ADT协同调控谱系转换的具体机制仍未阐明。本研究通过整合基于NEPC的CRISPR-Cas9筛选与肿瘤转分化的单细胞RNA测序（scRNA-seq）追踪分析，揭示了抗雄激素诱导的ZMYND8依赖性表观遗传编程调控NEPC转分化的核心机制。研究证实，在Pten/Trp53/Rb1敲除小鼠模型中，敲除Zmynd8可抑制NEPC的发生发展。反之，ASCL1介导的ZMYND8上调可重塑神经内分泌（NE）谱系，使肿瘤对雄激素受体（AR）靶向治疗产生耐药性。机制层面，去势抵抗性前列腺癌（CRPC）中的关键调控因子FOXM1，可稳定ZMYND8与携带H3K4me1-K14ac修饰及FOXM1结合位点的染色质区域的结合。抗雄激素治疗可使SWI/SNF染色质重塑复合物脱离AR，使其与ZMYND8/FOXM1结合，进而上调关键NE谱系调控因子（如FOXA2、SOX2及POU3F2的表达），最终诱导转分化过程。在明确CRPC患者中ZMYND8表达与不良预后的相关性后，本研究开发了一种可选择性抑制其组蛋白识别功能的小分子化合物ZMYND8i-34。在临床前模型中，ZMYND8i-34可有效阻断NE转分化并抑制CRPC进展。综上，本研究结果证实了抗雄激素治疗通过激活ZMYND8依赖性表观遗传重编程调控细胞谱系命运的关键作用，并提示靶向ZMYND8或可抑制NEPC发生并逆转治疗耐药性。整体实验设计：对过表达或未过表达ZMYND8的LNCap细胞进行染色质免疫共沉淀测序（ChIP-seq），分析ADT过程中ZMYND8过表达条件下ZMYND8结合位点的变化。