Core B: Bladder Cancer Models Core

项目摘要/摘要 我们的团队已经开发出新的方法来生成膀胱癌模型，该模型基于 人类患者来源的膀胱有机物和异种移植物，以及基因工程小鼠模型 (GEMMS)。特别是，我们为三维文化开发了一种创新的方法论 概括其相应亲本的组织病理学和分子特性的有机化合物 肿瘤，并具有人类膀胱癌特有的突变特征。此外，我们还开创了 肌肉浸润性膀胱癌(MIBC)的GEMM的发展，并已证明其在 联合临床研究。总之，这些资源为生成和分析 一系列体外和体内尿路上皮癌模型，将满足本计划项目的需要，如 以及更广泛的膀胱癌研究人员社区。 膀胱癌模型核心将通过以下方式支持我们计划项目的科学目标 建立人类和小鼠膀胱癌模型，这对所有三个项目都是至关重要的。核心是 围绕两个特定目标构建：在目标1中，我们将建立患者来源的膀胱癌的生物库 器官系，包括来自罕见膀胱癌亚型和特定基因组改变的患者 对方案项目的兴趣，以及妇女和少数民族患者的兴趣。通过与Core A合作，我们将 进行组织病理学和分子分析，以评估有机化合物与其 相应的亲本肿瘤，并将使用有针对性的外显子组测序来对它们的突变图谱进行分类。 我们的目标是建立一个代表膀胱癌全谱的器官样线生物库。 以及多样化的患者群体。所有三个项目都将使用这些有机管路，但将 对项目3尤为重要，该项目将研究患者的肿瘤异质性和克隆进化- 衍生的有机化合物。在目标2中，我们将产生和鉴定一系列膀胱癌的GEMM，包括 与计划项目特别相关的那些，即Kdm6a、ARID1a和Kmt2d，它们编码 在人类膀胱癌中频繁突变的表观遗传调节因子，以及核苷酸切除的Ercc2 与顺铂反应相关的修复途径基因。使用条件等位基因 对于这些基因中的每一个，我们将根据它们的功能丧失单独或在 与Trp53Flox/Flox；Ptenflx/Flox小鼠相结合，代表了MIBC的GEMM的良好特征。 我们将与Core A一起进行组织病理学和分子分析，以评估 这些GEMM对人类膀胱癌的治疗。这些GEM对项目1和2将是重要的，它们将 研究表观遗传调节因子在膀胱癌中的作用，对于项目3，补充 人体器官。最后，我们的工作将在本计划项目之外具有相当大的价值，通过提供 为更广泛的社区提供与临床相关的模式，促进新疗法的开发。

Project Summary/Abstract Our team has developed novel approaches for the generation of bladder cancer models based on human patient-derived bladder organoids and xenografts, as well as genetically-engineered mouse models (GEMMs). In particular, we have developed an innovative methodology for three-dimensional culture of organoids that recapitulate the histopathological and molecular properties of their corresponding parental tumors, and have mutational profiles characteristic of human bladder cancer. In addition, we have pioneered the development of GEMMs of muscle-invasive bladder cancer (MIBC), and have demonstrated their utility for co-clinical investigations. Together, these resources provide the foundation for the generation and analysis of a range of in vitro and in vivo models of urothelial cancer, which will serve the needs of this Program Project as well as the broader community of bladder cancer researchers. The Bladder Cancer Models Core will support the scientific objectives of our Program Project by generating human and mouse bladder cancer models that will be vital for all three Projects. The Core is structured around two specific aims: In Aim 1, we will establish a biobank of patient-derived bladder cancer organoid lines, including from patients with rare bladder cancer subtypes and genomic alterations of particular interest to the Program Project, and from women and minority patients. In collaboration with Core A, we will perform histopathological and molecular analyses to assess the similarity of the organoids to their corresponding parental tumors, and will use targeted exome sequencing to categorize their mutational profiles. Our goal is to generate a biobank of organoid lines that is representative of the full spectrum of bladder cancer as well as of a diverse patient population. These organoid lines will be utilized by all three Projects, but will be particularly important for Project 3, which will investigate tumor heterogeneity and clonal evolution in patient- derived organoids. In Aim 2, we will generate and characterize a series of GEMMs of bladder cancer, including those of particular relevance for the Program Project, namely Kdm6a, Arid1a, and Kmt2d, which encode epigenetic regulators that are frequently mutated in human bladder cancer, and Ercc2, a nucleotide excision repair pathway gene that is associated with cisplatin response. Using conditional alleles that have been obtained for each of these genes, we will generate GEMMs based on their loss-of-function alone or in combination with Trp53flox/flox; Ptenflox/flox mice, which represents a well-characterized GEMMs of MIBC. Together with Core A, we will perform histopathological and molecular analyses to assess the relationship of these GEMMs to human bladder cancer. These GEMMs will be important for Projects 1 and 2, which will investigate the functions of epigenetic regulators in bladder cancer, and for Project 3, to complement studies in human organoids. Finally, our work will be of considerable value beyond this Program Project by providing clinically-relevant models for the broader community that will facilitate development of new treatments.