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.

NUT 癌 (NC) 是一种由 BRD4-NUT 融合癌蛋白驱动的侵袭性鳞状癌。数控 影响所有年龄段，致死率很高 (>90%)，中位生存期为 6.5 个月。目前尚无有效治疗方法 NC 选项；因此，这种疾病代表了一种极端未得到满足的需求。该提案的总体目标是 通过机制驱动的治疗靶点识别和测试来提高这些患者的生存率。 为了满足对免疫活性动物模型的需求，其中 NC 细胞自主和非 可以严格研究自主致病机制，我们开发了第一个基因 NC 工程小鼠（GEM）模型。作为鳞状癌，NC 是融合的范例 癌蛋白驱动的实体瘤。 NC GEM 将扩展这一范式，以了解肿瘤内在的机制 和外部相互作用维持 NC 的增长。 从机制上讲，BRD4 是一种 BET 家族蛋白，其双溴结构域结合乙酰组蛋白，当融合时 NUT 会招募组蛋白乙酰转移酶 p300，形成称为巨域的巨大超级增强子。 BRD4-NUT 巨结构域维持促生长、抗分化转录因子的表达，包括 MYC、SOX2 和 TP63。我们的演示表明，使用 BET 溴结构域抑制剂 (BETi) 进行治疗，效果小 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，导致了研究 BRD4 在癌症中的作用的新领域。然而，已经很清楚的是 BETi 单一疗法并不能完全解决 NC 生物学问题。我们最近发现，抑制肿瘤 组蛋白甲基转移酶 EZH2 高度抑制基因表达，例如 CDKN2A/B 补充 BRD4-NUT 的致癌激活，维持 NC 生长。该途径的靶向 tazemetostat (taz) 与 BETi 具有高度协同作用，将在拟议的 GEM 模型中进行探索。 现在人们认识到 NC 具有免疫逃避的肿瘤微环境 (TME)，并且可以对 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，例如我们的 NC GEM 充分评估表观遗传修饰剂的作用以及免疫治疗在这种疾病中的作用。我们的创业板有一个 他莫昔芬诱导的鼠 Brd4 与人 NUTM1 条件性敲入融合，编码 BRD4-NUT 融合癌蛋白。我们的 NC GEM (‘mNC’) 中形成的侵袭性肿瘤提供了最明确的证据 BRD4-NUT 是这种癌症的唯一驱动因素。 mNC 非常模仿人类 NC，表现出快速增长， 转移性扩散，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 的组成也与人类 NC 相似。我们将利用我们新颖的 GEM 来解决 目标如下： 1. 建立 NC GEM (mNC) 对人类 NC (hNC) 生物学的适用性。 2. 设计 改善 NUT 癌的主要治疗。 3.探索预防NC复发的方法。