SHARPIN泛素化调控ASC/NLRP3诱发非可控性炎症促进前列腺癌增殖的机制研究

Nonresolving inflammation plays an important role in the pathogenesis of prostate cancer (PCa) and tumor-associated macrophages (TAM) can promote PCa proliferation, but the mechanisms are unknown.Our previous literature demonstrated that PCa prognostic factor SHARPIN is highly expressed in PCa and promote its progression. At the same time, the expression level of SHARPIN was positively correlated with the degree of TAM infiltration in PCa. Nevertheless, the relationship between SHARPIN and TAM is unclear. In the preliminary study, we found that SHARPIN activates NLRP3 inflammasome which can release inflammatory factors to regulate immune cells.Further studys showed that the SHARPIN modifies ASC,upstream protein of NLRP3,by ubiquitination, which may be related to the activation of NLRP3 inflammasome. Therefore, we hypothesized that SHARPIN can modify ASC by ubiquitination and promote its binding to NLRP3 to activate inflammasome. Then the activation of inflammasome would release inflammatory factors to raise activated TAM, induce nonresloving inflammation and promote PCa proliferation. Our project aims to clarify the key role of SHARPIN in the regulation of NLRP3 inflammasome and the specific mechanism of nonresolving inflammation in PCa through Co-IP, immunofluorescence and rescued experiments which hopes to provide the basis for the diagnosis and treatment of prostate cancer.

非可控性炎症在前列腺癌(PCa)发病具有重要作用，肿瘤相关巨噬细胞(TAM)可促进PCa增殖，然而机制未明。我们前期文章证明PCa预后相关因子SHARPIN在PCa高表达并促进其发展。同时SHARPIN表达水平与PCa中TAM浸润程度正相关，那么SHARPIN与TAM的调控关系如何？预实验发现SHARPIN可激活NLRP3炎症小体，而激活的炎症小体可释放炎性因子调控免疫细胞。进一步实验首次发现SHARPIN泛素化修饰NLRP3上游蛋白ASC，这可能与其激活NLRP3炎症小体有关。据此我们提出假设SHARPIN通过泛素化修饰ASC，促进其与NLRP3结合而激活炎症小体，释放炎性因子募集活化TAM，诱发非可控性炎症，促进PCa增殖。本项目拟通过Co-IP、免疫荧光和补救实验等技术阐明SHARPIN调控NLRP3炎症小体的关键作用及非可控性炎症在PCa中的具体机制，为其诊治提供理论依据。

我们通过免疫组化实验、体外功能实验、WB实验及动物实验证实SHARPIN可激活NLRP3炎症小体从而释放炎性因子调控免疫细胞，并发表文章证明PCa预后相关因子SHARPIN在PCa高表达并促进其发展。后续过测序及生信分析方式筛选出一批影响前列腺癌预后及临床病理特点的基因，包括MBTD1、PLK3、RAI14、ZC3H15等，并结合临床分析目的基因与预后关系。我们通过功能实验、动物模型验证其促转移功能，并通过转录组测序、IP、蛋白质谱等实验方式探究可能的作用机制。我们的研究成果阐明MBTD1、PLK3、RAI14、ZC3H15均为促癌基因，并促进前列腺癌骨转移，另外我们探索了其发挥作用的可能机制，为寻找针对PCa新的诊断及治疗靶点提供科学依据。