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

标题：开发临床相关的NUT癌基因工程小鼠模型 项目摘要： 睾丸特异性基因称为睾丸核蛋白（NUTM 1）已成为一个经常性的融合伴侣， 致癌融合基因在了解甚少的肿瘤中的作用在这些肿瘤中，NUT癌（NC）是最常见的肿瘤。 最具侵袭性，是NUTM 1融合基因相关癌症研究的范例。大多数 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（BRD 4）和NUTM 1蛋白。临床研究和实验研究基于在 体外细胞培养系统已经建立了BRD 4-NUTM 1融合蛋白作为NC的唯一驱动因子。这个项目 将解决NC研究中的两个重要问题： 问题1：奇怪的是，虽然BRD 4-NUTM 1足以驱动NC，但BRD 4-NUTM 1在NC中的异位表达， 非NC细胞具有明确的细胞毒性。是否存在NC起源的特定细胞类型-以及如何起源 这些细胞能够规避BRD 4-NUTM 1-的细胞毒性作用是一个谜。 问题2：作为NC肿瘤发生的驱动因素，BRD 4-NUTM 1是治疗NC最有前途的药物靶点。 然而，针对BRD 4-NUTM 1的BRD 4片段的靶向治疗策略已经被广泛应用。 令人失望的是，这主要是由于内源性BRD 4广泛地被 表达。靶向睾丸特异性NUTM 1蛋白有可能规避大多数系统性的方面， 男性患者的所有副作用和女性患者的所有副作用，但这种策略尚未在一个 生理背景 利用我们在基因组编辑技术方面的专业知识和基因工程小鼠的独特资源 我们将通过建立第一个基因工程小鼠模型来解决这两个重要的NC问题， 可以概括NC的组织和生理背景。目标1将建立和表征一个诱导型 染色体易位小鼠模型。我们的模型小鼠将表达用于追踪的报告基因， 分离表达BRD 4-NUTM 1的NC细胞。目的2通过对NC细胞的基因组分析，确定NC细胞的来源类型 来自NC来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BRD 4-NUTM 1蛋白降解以模拟NUTM 1靶向，以便建立靶向 以最小的副作用治疗NC的策略。 整体影响。该项目将产生一个基因工程小鼠模型的坚果癌作为一个完整的 研究BRD 4-NUTM 1致癌机制的新实验工具。我们的研究将改变 了解早期NC和刺激治疗发展，以治疗NC而无不可耐受的副作用。 该项目还将阐明其他鲜为人知的癌症的机制和治疗策略 NUTM 1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.