Genetically engineered mouse model to improve therapy of NUT carcinoma

基因工程小鼠模型可改善 NUT 癌的治疗

• 批准号: 10773306
• 负责人: Christopher A French
• 金额: $ 67.25万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-19 至 2028-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10773306
• 关键词: Acceleration; Address; Affect; Age; Animal Model; Basic Science; Binding; Biology; Bromodomain; Bromodomains and extra-terminal domain inhibitor; CD4 Positive T Lymphocytes; CD8B1 gene; CDKN2A gene; Carcinoma; Cells; Chromatin; Color; Communities; Complement; Credentialing; Disease; EP300 gene; EZH2 gene; Enhancers; Epigenetic Process; Ewings sarcoma; Excision; Fusion Oncogene Proteins; Gene Activation; Gene Expression; Gene set enrichment analysis; Genetic; Genetically Engineered Mouse; Goals; Growth; Histones; Histopathology; Human; Immune; Immune Evasion; Immune system; Immunocompetent; Immunofluorescence Immunologic; Immunologics; Immunophenotyping; Immunotherapy; Knock-in; Luciferases; Maintenance; Malignant Epithelial Cell; Malignant Neoplasms; Measures; Minority; Modeling; Mus; No Evidence of Disease; Oncogenic; Operative Surgical Procedures; Outcome; Pathogenicity; Pathway interactions; Patients; Population; Pre-Clinical Model; Protein Family; Proteomics; Repression; Role; Solid Neoplasm; Squamous cell carcinoma; T-Cell Depletion; T-Cell Receptor; Tamoxifen; Translational Research; Tumor Antigens; Tumor Suppressor Genes; Vaccination; burden of illness; chromatin remodeling; defined contribution; design; effective therapy; experimental study; histone acetyltransferase; human disease; human imaging; immune checkpoint blockade; immunomodulatory therapies; improved; novel; pre-clinical; protein complex; rapid growth; recruit; relapse prevention; response; small molecule; therapeutic development; therapeutic evaluation; therapeutic target; transcription factor; transcriptome; transcriptome sequencing; transcriptomics; tumor; tumor growth; tumor microenvironment; tumor-immune system interactions

NUT carcinoma (NC) is an aggressive squamous carcinoma driven by the BRD4-NUT fusion oncoprotein. NC affects all ages and is highly lethal (>90%), a median survival of 6.5 months. There are no effective treatment options for NC; thus, this disease represents an extreme unmet need. The overarching goal of this proposal is to improve survival of these patients through mechanism-driven identification and testing of therapeutic targets. In response to the need for an immunocompetent animal model in which both NC cell-autonomous and -non- autonomous pathogenic mechanisms can be investigated rigorously, we have developed the first genetically engineered mouse (GEM) model of NC. As a squamous carcinoma, NC serves as a paradigm for fusion oncoprotein-driven solid tumors. The NC GEM will expand that paradigm to understanding how tumor intrinsic and extrinsic interactions sustain NC growth. Mechanistically, BRD4 is a BET family protein whose dual bromodomains bind acetyl-histones that when fused to NUT, recruits the histone acetyltransferase, p300, forming enormous super-enhancers called megadomains. BRD4-NUT megadomains maintain expression of pro-growth, anti-differentiation transcription factors including MYC, SOX2, and TP63. Our demonstration that treatment with BET bromodomain inhibitors (BETi), small molecules that competitively inhibit binding of BET bromodomains to chromatin, can inhibit growth of NC in humans, led to a new field investigating the role of BRD4 in cancer. However, it has become clear that monotherapy with BETi does not fully address NC biology. We have recently found that repression of tumor suppressor gene expression, such as that of CDKN2A/B, by the histone methyltransferse, EZH2, highly complements oncogenic activation by BRD4-NUT in maintaining NC growth. Targeting of this pathway with tazemetostat (taz) is highly synergistic with BETi and will be explored in the proposed GEM model. It is now recognized that NC harbors an immune-evasive tumor microenvironment (TME) and can respond to immune modulation therapy. Epigenetic modulators such as taz and BETi are known to promote an anti-tumor immune TME. Thus, pre-clinical animal models with intact immune systems such as our NC GEM are needed to fully evaluate effects of epigenetic modifiers, and the role of immune therapy in this disease. Our GEM has a tamoxifen-inducible, conditional knock-in fusion of murine Brd4 with human NUTM1, encoding a BRD4-NUT fusion oncoprotein. Invasive tumors formed in our NC GEM (`mNC') have provided the most definitive evidence that BRD4-NUT is the sole driver of this cancer. mNC closely mimics human NC, demonstrating rapid growth, metastatic spread, and an indistinguishable histopathology and immunophenotype. Moreover, the immune cell composition of the mNC TME also resembles that of human NC. We will make use of our novel GEM to address the following aims: 1. establish the applicability of the NC GEM (mNC) to human NC (hNC) biology.; 2. devise improved primary therapy for NUT carcinoma.; 3. explore approaches to prevent relapse of NC.

坚果癌（NC）是由BRD4-NUT融合癌蛋白驱动的侵略性鳞状癌。 NC 影响所有年龄段，高度致命（> 90％），中位存活率为6.5个月。没有有效的治疗 NC的选项；因此，这种疾病代表了极端的未满足需求。该提议的总体目标是 通过机制驱动的鉴定和对治疗靶标的测试来提高这些患者的存活。 响应于需要免疫功能的动物模型，其中NC细胞自治和-Non-- 可以严格研究自主致病机制，我们已经开发了第一个遗传学 NC的工程鼠标（GEM）模型。作为鳞状癌，NC充当融合范式 癌蛋白驱动的实体瘤。 NC GEM将扩大该范式以了解肿瘤的内在 外部互动维持NC的增长。 从机械上讲，BRD4是一种BET家族蛋白 为了螺母，招募组蛋白乙酰转移酶P300，形成了巨大的超级增强剂，称为Megadomains。 Brd4-nut Megadomain保持促增长的抗分化转录因子的表达 MYC，SOX2和TP63。我们的证明表明，BET BET溴结构域抑制剂（BETI）的治疗，小 竞争性抑制BET溴结构域与染色质结合的分子可以抑制NC的生长 人类导致了一个新的领域，研究了BRD4在癌症中的作用。但是，很明显 BETI单一疗法不能完全解决NC生物学。我们最近发现抑制肿瘤 组蛋白甲基转化器EZH2，高度 通过BRD4-NUT在维持NC生长方面的致癌激活补充。用这个途径定位 TazeMetostat（TAZ）与BETI高度协同，将在拟议的GEM模型中进行探讨。 现在已经认识到，NC拥有免疫渗透肿瘤微环境（TME），并且可以回应 免疫调节疗法。已知表观遗传调节剂（例如TAZ和BETI）可以促进抗肿瘤 免疫TME。因此，需要具有完整免疫系统（例如我们的NC GEM）的临床前动物模型 完全评估表观遗传修饰剂的作用以及免疫治疗在该疾病中的作用。我们的宝石有一个 他莫昔芬可诱导的，有条件的鼠brd4与人Nutm1的融合，编码Brd4-nut 融合癌蛋白。在我们的NC GEM（“ MNC”）中形成的侵入性肿瘤提供了最明确的证据 该BRD4-NUT是该癌症的唯一驱动力。 MNC紧密模拟人类NC，证明快速生长， 转移扩散，以及无法区分的组织病理学和免疫表型。此外，免疫细胞 MNC TME的组成也类似于人类NC的组成。我们将利用我们的新颖宝石来解决 以下目的：1。建立NC GEM（MNC）对人NC（HNC）生物学的适用性。 2。设计 改善了坚果癌的一级治疗。 3。探索防止NC复发的方法。