基于单细胞RNA测序研究前列腺基质细胞异质性及Zeb1+基质亚群在前列腺上皮发育和癌变中的作用

肿瘤周围的基质细胞是前列腺癌发生发展的重要调节因素，然而基质细胞内部存在很大的异质性，对肿瘤的调控作用错综复杂。我们对前列腺肿瘤基质细胞的认识还亟待深入。申请人预实验发现：①单细胞RNA-seq提示，在前列腺发育和癌变时基质细胞的亚群组成和表达谱发生动态变化，其中Zeb1+亚群随发育的进行比例显著降低，在癌变后比例升高；②基质细胞中表达Zeb1促进前列腺肿瘤类器官和移植瘤的生长；③Zeb1+基质细胞可能是通过其富集表达的分泌蛋白Grem1发挥促瘤作用；④抗Grem1抗体具有显著的抑癌能力。我们将采用单细胞RNA-seq，利用基因工程小鼠、前列腺类器官、病人样本及移植瘤进行体内外实验，分析前列腺基质细胞的亚群组成，及其在发育和癌变过程中的变化，并深入研究Zeb1+基质细胞亚群在正常前列腺发育和肿瘤进展中的作用和机制，力图解析前列腺肿瘤基质细胞异质性，并为临床治疗提供新的方向。

Stromal cells are important regulatory elements in prostate cancer development and progression. However, our understanding on prostate stromal cells are far from complete due to their great heterogeneity and their complicated impacts on tumor cells. Our preliminary results indicate that (1) Single-cell RNA sequencing reveals that the mouse prostate stromal cells display dynamic changes as the mouse prostate develops or upon oncogenic transformation of epithelial cells. Particularly, a Zeb1 expressing subpopulation decreases as the prostate develops whereas increases in the prostate tumor tissue. (2) Zeb1 expression in stromal cells facilitate the growth of prostate tumor organoid and tumor xenograft. (3) The tumor-promoting effect of Zeb1+ stromal cell may be dependent on its secreted protein Grem1. (3) Anti-Grem1 antibody displays a significant inhibitory effect on prostate tumor organoid forming and tumor xenograft growth. We propose to investigate the dynamic change of stromal cell subset and their expression profile in prostate development and tumorigenesis utilizing single-cell RNA sequencing and data analyses. In addition, we will further explore the role and mechanism of Zeb1+ stromal subpopulation on prostate development and tumor progression using genetic engineered mouse models, the organoid culture system, patient samples and their derived xenograft. This study will provide new insight into the mechanism of prostate tumorigenesis and new clues for the development of novel therapeutic approaches.