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.

坚果癌是一种由BRD4-NUT融合癌蛋白驱动的侵袭性鳞癌。NC 影响所有年龄段，具有高度致命性(90%)，中位生存期为6.5个月。没有有效的治疗方法 NC的选择；因此，这种疾病代表着一种极端未得到满足的需求。这项提案的首要目标是 通过机制驱动的识别和治疗靶点的测试来提高这些患者的存活率。 响应于对一种免疫活性动物模型的需求，在该动物模型中NC细胞自主和非 自主致病机制可以严密研究，我们开发了第一个遗传学 NC的工程化小鼠模型。作为鳞癌，北卡罗来纳州是融合的典范 癌蛋白驱动的实体肿瘤。NC GEM将扩展这一范例，以了解肿瘤如何内在 外在的相互作用支持NC的增长。 从机制上讲，BRD4是一个BET家族蛋白，它的双溴结构域与乙酰组蛋白结合，当融合时 对于NUT，他招募了组蛋白乙酰转移酶p300，形成了巨大的超级增强剂，称为大核蛋白。 BRD4-NUT巨噬细胞维持促生长、抗分化转录因子的表达，包括 MYC、SOX2和TP63。我们的证据表明，使用BET溴域抑制剂(Beti)，小剂量 竞争性抑制BET溴结构域与染色质结合的分子，可抑制NC在 人类，导致了一个新的领域，研究BRD4在癌症中的作用。然而，情况已经变得很明显， 使用Beti的单一疗法不能完全解决NC生物学问题。我们最近发现对肿瘤的抑制 组蛋白甲基转移酶EZH2高度抑制CDKN2A/B等抑制基因的表达 补充BRD4-NUT在维持NC生长方面的致癌激活。以这条路径为靶点 TazemTostat(Taz)与Beti具有高度的协同性，并将在拟议的创业板模型中进行探索。 现已认识到，NC具有免疫逃逸的肿瘤微环境(TME)，并可对 免疫调节疗法。表观遗传调节剂，如taz和beti，已知可促进抗肿瘤 免疫TME。因此，需要具有完整免疫系统的临床前动物模型，如我们的NC GEM 充分评估表观遗传修饰物的效果，以及免疫治疗在本病中的作用。我们的宝石有一个 他莫昔芬诱导的小鼠Brd4与人NUTM1的有条件敲入融合，编码BRD4-NUT 融合癌蛋白。在我们的NC GEM(‘mNC’)形成的侵袭性肿瘤提供了最确凿的证据 BRD4-NUT是这种癌症的唯一驱动因素。MNC与人类NC非常相似，显示出快速增长， 转移扩散，以及难以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。我们将利用我们的新宝石来解决 目的如下：1.建立NC GEM(MNC)对人类NC(HNC)生物学的适用性；2.设计 改善NUT癌的初步治疗；3.探索预防NC复发的方法。