TRPM7的激酶域介导组蛋白H3修饰促进去势抵抗性前列腺癌生长和转移的机制研究

去势抵抗性前列腺癌（CRPC）发病机制复杂，治疗棘手。申报者前期报道，具有α-激酶结构域的TRPM7离子通道在CRPC中表达增高，促进CPRC生长和转移，与患者预后不良相关，但机制尚未阐明。预实验提示：TRPM7被剪切释放的C端激酶域（M7CK）是调控CRPC生长和转移的关键分子，M7CK可直接结合组蛋白H3并磷酸化H3S10、H3S28和H3T11位点，上调H3K4甲基化和H3K9乙酰化。因此，我们提出假说：TRPM7的激酶域片段核转位进入细胞核，磷酸化组蛋白H3，改变组蛋白整体修饰模式，调控下游基因转录，促进CRPC生长和转移。本项目拟采用CRISPR/Cas9、基因沉默和过表达、点突变、二代测序以及染色质免疫沉淀-qPCR等技术，获得M7CK直接磷酸化组蛋白H3调控基因转录的证据，并且通过体内外实验阐明M7CK促进CRPC生长和转移的表观遗传学机制，为CRPC的治疗提供新靶点。

The pathogenesis of castration-resistant prostate cancer (CRPC) is complicated, and the therapeutic approach for CRPC is limited. We previously reported that TRPM7, an ion channel with an α-kinase domain, was overexpressed in CRPC cells and tissue, and promoted the growth and metastasis of CRPC. Besides, we also found that the up-regulation of TRPM7 was closely correlated with the poor prognosis of CRPC patients. However, the molecular mechanism remains unclear. Our preliminary experiments showed that TRPM7-cleaved kinase (M7CK) was released from the C-terminal of TRPM7 and played an essential role on the growth and metastasis of CRPC. The results showed that M7CK could directly bind to histone H3 and phosphorylate H3S10, H3S28, and H3T11, and up-regulate H3K4 methylation and H3K9 acetylation. Therefore, we hypothesize that M7CK translocates into the nucleus, phosphorylates histone H3, and then changes the global modification pattern of histone , which subsequently regulates the gene transcription accounting for the growth and metastasis of CRPC. Our project will employ multiple experimental techniques including CRISPR/Cas9, gene silence and overexpression, gene site-directed mutation, next generation sequencing (NGS), and chromatin immunoprecipitation-qPCR (ChIP-qPCR) to prove that M7CK directly phosphorylate histone H3 to regulate gene transcription. We will further carried out both in vitro and in vivo experiments to elucidate the epigenetic mechanism by which M7CK promotes the growth and metastasis of CRPC. This study will provide a novel target for the treatment of CRPC.