in vivo live imaging and Single cell gene expression analysis of macrophages during tumour initiation

肿瘤发生过程中巨噬细胞的体内实时成像和单细胞基因表达分析

• 批准号: 2097905
• 金额: --
• 依托单位: University of Edinburgh
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2018
• 资助国家: 英国
• 起止时间: 2018 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2097905
• 关键词: vivo; live; imaging; Single; cell

The development and progression of cancer involve a complex interplay between pre- neoplastic cells (PNCs) and the host environment. Until recently, it was not possible to live image the earliest interactions between host cells and PNCs during tumour initiation. Earlier work in the Feng lab, using zebrafish as a model organism, has made it possible to visualize a PNC at its inception in vivo [1]. This work revealed that a PNC elicits a Trophic Inflammatory response in which recruited innate immune cells including neutrophils and macrophages promote PNC growth, at least in part, through COX-2 mediated PGE2 production [1]. This has immediately provided us with a potential target for cancer prevention.Our more recent in vivo live imaging work using several activation reporter fish (NFkB, TNFa, Irg1) and macrophage reporter fish (mpeg1, mfap4) revealed that recruited macrophages within PNC developing niche are highly heterogeneous. Evidence from the literature suggests that macrophage subpopulations can inter-convert within inflamed tissue during the course of an inflammation response. Within the PNC developing niche, the particular balance among macrophage states with different phenotypes might have a major impact on the final outcome of PNC progression. Therefore we need to characterize better the complete spectrum of macrophages and their states within the PNC developing niche and to define their roles during PNC progression.Single cell RNA-sequence analysis is a powerful technique with the potential to systematically map all macrophage phenotypes within the PNC niche in an unbiased manner. This would reveal rare populations and assign developmental relations among different cellular phenotypes. We (YF/CPP) have already applied this technique to characterize neutrophils within the PNC niche, and identified a unique but rare neutrophil population that could be key to PNC promotion.Here we propose to use a recently established inducible zebrafish PNC model [2] for single cell RNA-seq analysis of macrophages isolated from the PNC niche and in vivo live imaging studies of PNC associated macrophages. This will allow us to characterize PNC promoting macrophage and to elucidate how the PNC promoting phenotype is established. A likely outcome of this project is the identification of novel targets for cancer prevention that modulate the interplay between host immune cell and PNC.

肿瘤的发生和发展涉及肿瘤前细胞（pnc）与宿主环境之间复杂的相互作用。直到最近，还不可能实时成像肿瘤起始期间宿主细胞和pnc之间最早的相互作用。Feng实验室的早期工作，使用斑马鱼作为模型生物，使得在体内观察PNC的初始状态成为可能。这项工作揭示了PNC引发营养性炎症反应，其中招募的先天免疫细胞包括中性粒细胞和巨噬细胞，至少部分通过COX-2介导的PGE2产生促进PNC的生长。这立即为我们提供了预防癌症的潜在目标。我们最近使用几种激活报告鱼（NFkB, TNFa, Irg1）和巨噬细胞报告鱼（mpeg1, mfap4）进行的活体成像研究表明，PNC发育生态位中招募的巨噬细胞是高度异质性的。文献证据表明，在炎症反应过程中，巨噬细胞亚群可以在炎症组织内相互转化。在PNC发育生态位中，不同表型巨噬细胞状态之间的特殊平衡可能对PNC进展的最终结果产生重大影响。因此，我们需要更好地描述巨噬细胞的全谱及其在PNC发育生态位中的状态，并确定它们在PNC进展中的作用。单细胞rna序列分析是一项强大的技术，有可能以公正的方式系统地绘制PNC生态位内所有巨噬细胞表型。这将揭示罕见种群和分配不同细胞表型之间的发育关系。我们（YF/CPP）已经应用这种技术来表征PNC生态位中的中性粒细胞，并确定了一个独特但罕见的中性粒细胞群体，可能是PNC促进的关键。在这里，我们建议使用最近建立的诱导斑马鱼PNC模型[2]对从PNC生态位分离的巨噬细胞进行单细胞RNA-seq分析，并对PNC相关巨噬细胞进行活体成像研究。这将使我们能够表征PNC促进巨噬细胞，并阐明PNC促进表型是如何建立的。这个项目的一个可能的结果是确定新的癌症预防靶点，调节宿主免疫细胞和PNC之间的相互作用。