Analysis of epithelial heterogeneity in prostate development and cancer

项目摘要 启动癌症的细胞类型，或称“起源细胞”，在确定肿瘤的过程中起着关键作用。 性质，如分子和组织病理亚型以及治疗反应和疾病 结果。在前列腺癌的情况下，起源细胞可能与它是惰性的还是 侵袭性，因此对起源的细胞类型的分析可能导致识别 可以指导治疗。在过去的十年里，我们实验室在认识上取得了长足的进步 通过对祖细胞群体、血统关系和 前列腺上皮在正常发育过程中的转录调节以及癌症的发生和发展 进步。在这项提议的初步研究中，我们已经进行了单细胞RNA测序 检测良性小鼠和人前列腺中前列腺上皮细胞的异质性。值得注意的是，这些 分析揭示了几种不同的管腔种群，这些种群以前没有被发现，可能 代表不同来源的细胞类型。此外，我们还使用了计算系统方法来 确定这些新的管腔群体的候选主控调节(驱动)基因。最后，我们有 建立了三维类器官培养条件，以考察特定的小鼠和人 上皮细胞群是前列腺癌起源的细胞类型。 根据我们的初步数据，我们假设对特定原因和驱动因素的分析 前列腺癌的上皮细胞群将阐明前列腺癌起源的细胞类型及其作用。 在确定肿瘤表型方面。为了研究这一假设，我们将寻求一种创新的组合 使用基因工程小鼠模型的体内、体外、分子和计算系统方法 和人类的前列腺组织。我们提出了三个具体的目标：(1)研究上皮细胞的异质性 用单细胞转录组分析前列腺癌的起源和血统关系 不同的上皮细胞群体，并确定指定这些群体的驱动因素；(2)细胞起源的分析 在小鼠和人的前列腺中，使用有机物培养方法来确定不同的管腔和 癌变后基底上皮细胞群可诱发肿瘤及识别候选细胞 这一过程的主要调控因素；(3)小鼠和人上皮异质性的研究 前列腺癌使用跨物种单细胞分析来确定肿瘤异质性的保守驱动因素。 综上所述，我们的研究将提供对前列腺上皮细胞的基本分子和功能洞察 在发育和肿瘤发生过程中的异质性，将对理解 人类前列腺癌的起源及其治疗。

Project Summary The cell type that undergoes cancer initiation, or “cell of origin,” plays a key role in determining tumor properties such as molecular and histopathological subtype as well as treatment response and disease outcome. In the case of prostate cancer, the cell of origin may be highly relevant for whether it is indolent or aggressive, and consequently the analysis of cell types of origin may lead to identification of biomarkers that can guide therapy. Over the past ten years, our laboratory has made considerable progress in understanding prostate cell types of origin through studies of the progenitor populations, lineage relationships, and transcriptional regulators of the prostate epithelium during normal development as well as cancer initiation and progression. In preliminary studies for this proposal, we have performed single-cell RNA sequencing to examine the heterogeneity of prostate epithelial cells in the benign mouse and human prostate. Notably, these analyses have revealed several different luminal populations that have not been previously identified, and may represent distinct cell types of origin. Furthermore, we have used computational systems approaches to identify candidate master regulator (driver) genes for these novel luminal populations. Finally, we have established three-dimensional organoid culture conditions to investigate whether specific mouse and human epithelial cell populations are cell types of origin for prostate cancer. Based on our preliminary data, we hypothesize that analysis of the origin and drivers of specific epithelial populations in the prostate will elucidate cell types of origin for prostate cancer as well as their roles in specifying tumor phenotypes. To investigate this hypothesis, we will pursue an innovative combination of in vivo, ex vivo, molecular, and computational systems approaches using genetically-engineered mouse models and human prostate tissue. We propose three specific aims: (1) Investigation of epithelial heterogeneity in the developing and adult prostate using single-cell transcriptomics to examine the origin and lineage relationships of distinct epithelial populations and identify drivers that specify these populations; (2) Analysis of cell of origin in mouse and human prostate using organoid culture approaches to determine whether distinct luminal and basal epithelial populations can give rise to tumors after oncogenic transformation and identify candidate master regulators of this process; and (3) Investigation of epithelial heterogeneity in mouse and human prostate cancer using cross-species single-cell analyses to identify conserved drivers of tumor heterogeneity. Taken together, our studies will provide fundamental molecular and functional insights into prostate epithelial heterogeneity during development and tumorigenesis, and will have important implications for understanding the origins of human prostate cancer and its treatment.