Core B: Bladder Cancer Models Core

项目概要/摘要 我们的团队已经开发了新的方法，用于生成膀胱癌模型， 人类患者来源的膀胱类器官和异种移植物，以及基因工程小鼠模型 （GEMMs）。特别是，我们开发了一种创新的三维文化方法， 概括其相应亲本的组织病理学和分子特性的类器官 肿瘤，并具有人膀胱癌的突变特征。此外，我们还开创了 肌层浸润性膀胱癌（MIBC）的GEMM的发展，并已证明其在 联合临床研究。这些资源共同为生成和分析 一系列尿路上皮癌的体外和体内模型，这将满足本计划项目的需要， 以及更广泛的膀胱癌研究者群体。 膀胱癌模型核心将支持我们的计划项目的科学目标， 人类和小鼠膀胱癌模型，这将是至关重要的所有三个项目。核心是 围绕两个具体目标构建：在目标1中，我们将建立一个患者来源的膀胱癌生物库 类器官系，包括来自罕见膀胱癌亚型和特定细胞的基因组改变的患者。 该计划项目的利益，以及妇女和少数民族患者。与核心A合作，我们将 进行组织病理学和分子分析，以评估类器官与其 因此，本发明的方法将使用靶向外显子组测序来对相应的亲本肿瘤进行分类，并且将使用靶向外显子组测序来对其突变谱进行分类。 我们的目标是建立一个代表膀胱癌全谱的类器官细胞系生物库 以及不同的患者群体。这些类器官系将被所有三个项目利用，但将 对于项目3尤其重要，该项目将研究患者的肿瘤异质性和克隆进化， 衍生类器官。在目标2中，我们将生成并表征一系列膀胱癌的GEMM，包括 与计划项目特别相关的那些，即Kdm 6a、Arid 1a和Kmt 2d，它们编码 在人类膀胱癌中经常突变的表观遗传调节因子，以及Ercc 2，一种核苷酸切除， 与顺铂反应相关的修复途径基因。使用条件等位基因， 获得这些基因中的每一个，我们将产生GEMM的基础上，他们的功能丧失单独或 与Trp 53 flox/flox组合; Ptenflox/flox小鼠，其代表MIBC的良好表征的GEMM。 与核心A一起，我们将进行组织病理学和分子分析，以评估 这些GEMM对人类膀胱癌的作用。这些GEMM对项目1和2很重要， 研究表观遗传调节因子在膀胱癌中的功能，对于项目3，补充 人类类器官最后，我们的工作将通过提供 为更广泛的社区提供临床相关模型，以促进新疗法的开发。

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.