Developing a clinically-relevant genetically engineered mouse model for Nut carcinoma

标题：建立与临床相关的NUT癌基因工程小鼠模型 项目总结： 一种称为睾丸核蛋白(NUTM1)的睾丸特异基因已经成为 鲜为人知的肿瘤中的致癌融合基因。在这些肿瘤中，坚果癌(NC)是最常见的 是最具侵袭性的，是研究NUTM1融合基因相关癌症的范例。大多数人 NC与染色体易位有关，该易位将溴结构域的基因片段连接在一起- 含有蛋白4(BRD4)和NUTM1蛋白。基于智能网的临床研究和实验研究 体外细胞培养系统已经确定BRD4-NUTM1融合蛋白是NC的唯一驱动因素。这个项目 将解决NC研究中的两个关键当前问题： 问题1：矛盾的是，尽管BRD4-NUTM1足以驱动NC，但BRD4-NUTM1在 非NC细胞具有明确的细胞毒性。NCs是否存在特定的细胞类型--以及如何产生 这些细胞能够绕过BRD4-NUTM1-的细胞毒性效应是一个谜。 问题2：作为NC发生的驱动力，BRD4-NUTM1是治疗NC最有前景的药物靶点。 然而，针对BRD4-NUTM1的BRD4片段的靶向治疗策略已经被 令人失望，很大程度上是因为内源性BRD4广泛存在导致的无法忍受的副作用 表达。靶向睾丸特异性NUTM1蛋白有可能绕过大多数系统性方面 对男性患者的影响和对女性患者的所有副作用，但这一策略尚未在 生理背景。 以我们在基因组编辑技术和转基因小鼠独特资源方面的专业知识为基础 LINES，我们将通过建立第一个基因工程小鼠模型来解决这两个关键的NC问题 可以概括NC的组织和生理背景。目标1将构建和描述一个诱导性 NC小鼠染色体易位模型的建立。我们的模型小鼠将表达报告基因，用于追踪和 分离表达BRD4-NUTM1的NC细胞。目标2将通过分析基因组来鉴定起源的NC细胞类型 来自NC起源的正常细胞群体和NC发育早期阶段的细胞的数据。AIM 3将利用 体内可诱导的BRD4-NUTM1蛋白降解模拟NUTM1靶向以建立靶向 治疗NC的策略，副作用最小。 整体影响。该项目将生产一种基因工程的坚果癌小鼠模型，作为一种 研究BRD4-NUTM1致癌机制的新实验工具。我们的研究将会改变 了解早期NC并促进治疗发展，以治疗NC，而不会出现无法忍受的副作用。 该项目还将阐明其他知之甚少的癌症的机制和治疗策略。 携带NUTM1融合基因。

Title: Developing a Clinically-Relevant Genetically Engineered Mouse Model of NUT Carcinoma Project Summary: A testis-specific gene called Nuclear protein in testis (NUTM1) has emerged as a recurrent fusion partner of oncogenic fusion genes in poorly understood neoplasms. Among these neoplasms, NUT carcinoma (NC) is the most aggressive and is the paradigm for the study of NUTM1 fusion gene-associated cancers. The majority of NCs are associated with a chromosome translocation that joins together gene fragments of the Bromodomain- containing protein 4 (BRD4) and the NUTM1 protein. Clinical research and experimental research based on in vitro cell culture systems has established the BRD4-NUTM1 fusion protein as the sole driver of NC. This project will address two vital current issues in NC research: Issue 1: Paradoxically, although BRD4-NUTM1 suffices to drive NC, ectopic expression of BRD4-NUTM1 in non-NC cells is unequivocally cytotoxic. Whether there is a specific cell type in which NCs originate — and how these cells are able to circumvent the cytotoxic effects of BRD4-NUTM1 — is a mystery. Issue 2: As the driver of NC oncogenesis, BRD4–NUTM1 is the most promising drug target for treating NC. However, targeted therapy strategies directed against the BRD4 fragment of BRD4-NUTM1 have been disappointing, largely because of intolerable side effects that are due to endogenous BRD4 being broadly expressed. Targeting the testis-specific NUTM1 protein has the potential to circumvent most systematic side effects in male patients and all side effects in female patients, yet this strategy has not been explored in a physiological context. Building on our expertise in genome editing technology and unique resources in genetically engineered mouse lines, we will address these two vital NC issues by building the first genetically engineered mouse model that can recapitulate the tissue and physiological context of NC. Aim 1 will build and characterize an inducible chromosome translocation mouse model for NC. Our model mice will express reporter genes for tracing and isolating NC cells that express BRD4-NUTM1. Aim 2 will identify the NC cell type of origin by analyzing genomic data from NC-originating normal cell populations and cells in early stages of NC development. Aim 3 will leverage in vivo inducible BRD4-NUTM1 protein degradation to model NUTM1 targeting in order to establish a targeting strategy for treating NC with minimal side effects. Overall Impact. This project will produce a genetically engineered mouse model of Nut Carcinoma as an entirely new experimental tool for studying oncogenic mechanisms of BRD4-NUTM1. Our studies will transform understanding of early-stage NC and spur therapeutic development to treat NC without intolerable side effects. This project will also shed light on mechanisms and treatment strategies for other poorly understood cancers with NUTM1 fusion genes.