Role of histone demethylase KDM5C in metastatic prostate cancer

癌症转移是毁灭性的，临床上具有挑战性。对于转移，癌细胞获得了增强的迁移和侵袭能力，原则上受表观遗传控制。在我们之前的研究中，我们报道了组蛋白去甲基化酶KDM5C（赖氨酸去甲基化酶5C，也称为JARID1C或SMCX）在前列腺癌中过度表达，并且KDM5C是根治性前列腺切除术后前列腺特异性抗原复发的预后标志物。在这里，我们发现KDM5C在转移性前列腺癌中被高度上调，而KDM5C的下调导致前列腺癌细胞的迁移和侵袭减少。在该项目中，我们的目标是扩大对转移性前列腺癌中组蛋白去甲基化酶KDM5C的潜在分子机制和影响的了解。首先，与对照细胞相比，我们将通过对KDM5C缺失和/或KDM5C过表达的细胞进行RNA-Seq检测KDM5C是否会影响前列腺癌细胞。由此，我们将确定KDM5C可能影响肿瘤细胞进展的信号通路。其次，我们想通过ChIP-Seq实验确定KDM5C是否占据靶基因并在H3K4me3/2的启动子上去甲基化。基因表达分层和KDM5C结合的全基因组两两比较将确定直接的KDM5C靶基因。该结果将为KDM5C的下调或抑制如何增加转移相关基因的表达以增强前列腺癌细胞的侵袭能力提供表观遗传学见解。为了对kdm5c介导的基因调控有新的认识，我们希望寻找kdm5c相互作用的蛋白。亲和纯化和kdm5c相关蛋白的质谱分析将允许鉴定未知的kdm5c相互作用蛋白。该项目产生的数据将提高对转移性前列腺癌发展过程中发生的分子机制的认识，并将回答靶向KDM5C是否为减轻转移负担的策略的问题。

Cancer metastasis is devastating and clinically challenging. For metastasis, cancer cells acquire increased migratory and invasive abilities, which are in principle subject to epigenetic control. In our previous study, we reported that the histone demethylase KDM5C (lysine demethylase 5C, also called JARID1C or SMCX) is over-expressed in prostate cancer and that KDM5C is a prognostic marker for prostate-specific antigen-relapse following radical prostatectomy. Here, we show that KDM5C is highly up-regulated in metastatic prostate cancer and that down-regulation of KDM5C leads to decreased migration and invasion of prostate cancer cells. In the proposed project, we aim to expand the knowledge about the underlying molecular mechanism and the impact of the histone demethylase KDM5C in metastatic prostate cancer. Firstly, we will examine whether KDM5C influences prostate cancer cells by performing RNA-Seq of KDM5C-depleted and/or KDM5C over-expressing cells in comparison to control cells. By this we will determine the signaling pathway(s) through which KDM5C may affect tumor cell progression. Secondly, we want to determine if KDM5C occupies target genes and demethylates H3K4me3/2 at their promoters by performing ChIP-Seq experiments. Stratification of gene expression and genome-wide pairwise comparison of KDM5C binding will identify direct KDM5C target genes. The results will provide epigenetic insights into how the down-regulation or inhibition of KDM5C may increase expression of metastasis-linked genes to enhance the invasive ability of prostate cancer cells. To gain new insight into KDM5C-mediated gene regulation we want to search for KDM5C-interacting proteins. Affinity purification and a mass spectrometric analysis of KDM5C-associated proteins will allow identifying unknown KDM5C-interacting proteins. The data emerging from this project will improve the knowledge about the molecular mechanisms taking place during development of metastatic prostate cancer and will answer the question whether targeting KDM5C is a strategy to reduce metastatic burden.