P2: Modelling epigenetic tumour suppressor-driven urothelial carcinomas in mice

该项目旨在了解特定表观遗传肿瘤抑制基因的突变如何促进膀胱癌的早期发展，以及向浸润性癌症的进展。在第一批工作中，我们将使用遗传方法在小鼠膀胱尿路上皮中模拟人类膀胱癌发展的最初步骤。在第二个工作包中，我们将使用小鼠尿路上皮类器官培养作为实验系统来诱导组合基因突变，以模拟人类膀胱癌中出现的表观遗传肿瘤抑制基因和其他致癌途径中的复杂组合遗传改变。在第三个目标中，我们将开发一种新的实验系统，以允许快速生成小鼠系，这些小鼠系允许细胞类型特异性，强力霉素诱导，组合敲除或基因过表达，以便生成更准确的膀胱癌小鼠模型。所有这些模型系统将使用染色质结合、染色质修饰和RNA转录的一系列基因组尺度分析来研究，以获得表观遗传肿瘤抑制基因突变如何导致膀胱癌形成的分子洞察力。我们还将在化学文库和遗传文库筛选研究中使用我们的模型系统，以识别和测试利用表观遗传肿瘤抑制基因特定突变导致的脆弱性的新治疗机会。与所有其他UcarE项目的密切互动将允许将我们在小鼠研究中取得的发现转化为人类膀胱癌样本，类器官和细胞系。在这个项目中产生的各种实验性膀胱癌模型系统将反过来对UcarE的其他几个项目非常有用，这些项目可以通过研究小鼠膀胱生理背景下的人类细胞系统来研究见解，并将为临床前治疗测试提供实验模型。

This project aims to understand how mutations in specific epigenetic tumour suppressor genes contribute to the earliest stages of development of bladder cancer, as well as to progression to invasive cancer. In a first package of work we will use genetic approaches to mimic the earliest steps of human bladder cancer development in the mouse bladder urothelium. In the second work package we will induce combinatorial genetic mutations using mouse urothelial organoid cultures as an experimental system in order to mimic the complex combinatorial genetic alterations in epigenetic tumour suppressor genes and in other oncogenic pathways that arise in human bladder cancer. In a third aim, we will develop a new experimental system to allow the rapid generation of lines of mice that allow the cell type-specific, doxycycline-inducible, combinatorial knockout or overexpression of genes in order to generate more accurate mouse models of bladder cancer. All of these model systems will be studied using a series of genome-scale analyses of chromatin binding, chromatin modification and RNA transcription to gain molecular insight into how mutations in epigenetic tumour suppressor genes lead to bladder cancer formation. We will also use our model systems in chemical library and genetic library screening studies to identify, and test, new therapeutic opportunities that exploit vulnerabilities resulting from specific mutations in epigenetic tumour suppressor genes. Close interactions with all of the other UcarE projects will permit translation of the findings made in our mouse studies to human bladder cancer samples, organoids and cell lines. The diverse experimental bladder cancer model systems that will be generated in this project will in turn be very useful for several other UcarE projects to allow investigation of insights made through the study of human cellular systems in the physiological context of the mouse bladder, and will provide experimental models for pre-clinical therapeutic testing.