基于单细胞转录组测序的垂体腺瘤增殖异质性评价及与侵袭表型相关性研究

项目名称： 基于单细胞转录组测序的垂体腺瘤增殖异质性评价及与侵袭表型相关性研究

项目编号： No.31671109

项目类型： 面上项目

立项/批准年度： 2017

项目学科： 神经、认识与心理学

项目作者： 周大彪

作者单位： 首都医科大学

项目金额： 25万元

中文摘要： 目前研究对垂体腺瘤细胞的谱系分化与功能表型所知甚少，肿瘤内异质性即基因学不同的亚克隆群调控疾病进程。我们前期工作发现增殖相关基因表达促进肿瘤进展，以及垂体腺瘤增殖标志物表达和超微结构的异质性。本课题以复发垂体腺瘤和原发侵袭性垂体腺瘤为研究对象，应用高通量单细胞RNA测序技术分析肿瘤个体细胞在转录组水平的基因型表达，着重于发现与细胞增殖相关的差异表达基因，应用生物信息学方法识别具有共同表达谱特征的细胞亚群，同时探索已知的增殖相关基因AURKA和NDC80与新型的细胞亚群标记基因作为生物学标志物与垂体腺瘤侵袭表型的相关性，并在核酸和蛋白水平验证；更为重要的是，通过转录组特征分析可能发现肿瘤内部具有干细胞特性的细胞亚群及其谱系分化特性。预期研究结果将建立基于单细胞转录组数据的垂体腺瘤亚型分型，获得肿瘤细胞亚群的增殖异质性与肿瘤侵袭和进展相关的直接证据。

中文关键词： 垂体腺瘤；单细胞RNA测序；异质性；增殖；转录组

英文摘要： At present, little is known about the lineage differentiation and functional phenotype of the pituitary adenoma cells. Intratumoral heterogeneity that is genetically different subclones control disease progression. Our preliminary work revealed that proliferation-related gene expression promotes tumor progression, as well as the heterogeneity of the proliferative marker expression and ultrastructure of pituitary adenomas. The recurrent and primary pituitary adenomas with invasive subtypes were enrolled in the present research. We analyze gene expression of individual tumor cells at the transcriptome level using high-throughput single-cell RNA-seq. We aim to (1) explore the differentially expressed genes related to cell proliferation; (2) identify the cell subpopulation with common expression profiles by bioinformatics analysis and (3) explore the correlation between the expression of AURKA/NDC80 and the new subpopulation marker genes as biomarkers and the invasive phenotype of pituitary adenomas, and further to validate at the level of mRNA and protein. More important, the cell subpopulations with characteristics of stemness and their lineage differentiation could be identified by the transcriptional signatures analysis. The results of the study are expected to establish the subtype classification of pituitary adenoma based on single-cell transcriptome data, and to obtain the direct evidence that the proliferation heterogeneity of individual cell subpopulation causes tumor invasion and progression.

英文关键词： pituitary adenoma;Single-cell RNA-Seq;heterogeneity;proliferation;transcriptome