中心体扩增影响前列腺癌发生与发展的机制研究

Centrosome amplifications, the hallmarks of human cancer, can induce the chromosomal instability, disturb the integrity of tissue homeostasis and trigger cell invasions. Several in vitro and in vivo studies have shown that centrosome amplifications can promote the tumorigenesis and tumor progression. Clinical evidences also indicate that centrosome amplifications are correlated with the poor diagnosis and malignancy of human tumors. Previously, we have shown that centrosome amplifications can promote the formation of prostatic intraepithelial neoplasia (PIN) in a transgenic mouse model. However, it is still unknown whether and how centrosome amplifications affect the tumorigenesis and/or tumor progression of the prostate. In this project, we propose to study the molecular and cellular mechanisms underlying the centrosome amplification-elicited tumor formation through specifically inducing the centrosome amplifications in the mouse prostate epithelium. In addition, we will use a genetic strategy that randomly induces centrosome amplifications on individual tumor cells and trace these cells over time during the relapse and metastasis of castration-resistant prostate tumors. Furthermore, we will perform a high-throughput screening to identify molecules that could induce the centrosome amplification in the prostate epithelium and to identify the tumor-related signaling pathways elicited by the centrosome amplification as well as the critical molecule involved. Finally, we will determine the function of these molecules on human prostate tumorigenesis and tumor progression by using clinical prostate tumor tissues and cancer cell lines. These studies will facilitate us to understand whether and how centrosome amplifications affect the tumorigenesis and/or tumor progression of the prostate epithelium and identify molecules that could prevent or treat the human prostate cancer.

中心体数目扩增是肿瘤细胞的特征之一。中心体扩增能够诱导染色体不稳定、破坏组织稳态、促进细胞的浸润与侵袭，具有促进肿瘤发生发展的能力。我们前期在转基因小鼠的研究发现中心体扩增能够促进小鼠前列腺上皮内瘤的生成。然而，中心体扩增影响前列腺肿瘤发生发展的机制尚不明确。本研究利用转基因小鼠，在体内特异性地诱导前列腺上皮中心体扩增，探讨其通过何种细胞与分子机制影响前列腺癌的发生与发展；并通过转基因，在体内随机诱导单个前列腺肿瘤细胞中心体扩增、追踪这些肿瘤细胞在去雄性激素治疗后肿瘤复发转移过程中的作用；继而通过cDNA芯片从转基因小鼠中筛选诱导前列腺上皮中心体扩增的重要分子、中心体扩增激活的肿瘤相关信号转导通路、以及这些通路上的关键分子；并在临床前列腺癌组织上验证这些分子对人前列腺癌发生发展的影响。本项目的完成可明确中心体扩增影响前列腺癌发生发展的作用及其机制并可为临床前列腺癌的诊断与治疗提供新的靶点。

本项目通过成功构建去势抵抗前列腺癌神经内分泌样分化转基因小鼠模型，明确了染色体不稳定与中心体扩增促进前列腺癌的进展，并与去势抵抗前列腺癌产生神经内分泌样分化/小细胞癌呈正相关。通过建立神经内分泌样分化前列腺肿瘤类器官培养体系，获得多株去势抵抗前列腺神经内分泌样分化肿瘤类器官株，并将此技术推广到其他肿瘤类型；继而采用单细胞测序对体外培养类器官细胞分型，发现类器官模拟恶性肿瘤由雄激素受体阳性腺瘤，逐渐降低雄激素受体表达并进展为神经内分泌样分化肿瘤，直至雄激素受体表达阴性形成小细胞癌的全过程；随后利用单细胞ATAC-Seq技术发现肿瘤类器官具有可塑性，利用Chip-Seq技术发现了诱导去势抵抗前列腺癌进展为神经内分泌样分化/小细胞癌的表观遗传调控机制；最后构建神经内分泌样分化肿瘤类器官筛药平台，利用VIPER生物信息学技术进行双药或多药预测，并对不同的药物组合进行验证，发现多对有效控制神经内分泌样分化肿瘤生长的药物组合。此次研究为去势抵抗前列腺癌神经内分泌样分化/小细胞癌构建小鼠模型；揭示了影响去势抵抗前列腺癌细胞可塑性的表观遗传调控机制；并建立可用于药物筛选的神经内分泌样分化类器官株与药物筛选平台。研究成果已有一篇发表于《Cold Spring Harbor Perspectives in Medicine》上； 另外两篇分别投稿《Nature》与《Nature Communications》杂志，正予以评审；推广神经样分化肿瘤类器官培养技术的文章投稿《Diseases of the Colon & Rectum》，正予以评审。