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）是由BRD 4-NUT融合癌蛋白驱动的侵袭性鳞状细胞癌。NC 影响所有年龄段，并且是高度致命的（>90%），中位生存期为6.5个月。没有有效的治疗方法 因此，这种疾病代表了一种极端未满足的需求。本提案的总体目标是 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生存率。 为了响应对免疫活性动物模型的需要，其中NC细胞自主的和非自主的都是免疫活性动物模型。 自主致病机制可以严格调查，我们已经开发出第一个基因 NC的工程小鼠（GEM）模型。作为一种鳞状细胞癌，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 和外部相互作用维持NC生长。 在机制上，BRD 4是BET家族蛋白，其双溴结构域结合乙酰基-组蛋白，当融合时， 到NUT，招募组蛋白乙酰转移酶，p300，形成巨大的超级增强子，称为megadomains。 BRD 4-NUT巨蛋白维持促生长、抗分化转录因子的表达，包括 MYC、SOX 2和TP 63。我们证明，BET溴结构域抑制剂（BETi）治疗，小 竞争性抑制BET布罗莫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BRD 4在癌症中的作用。然而，很明显， 使用BETi的单一疗法不能完全解决NC生物学。我们最近发现抑制肿瘤 组蛋白甲基转移酶EZH 2对抑制基因表达，如CDKN 2 A/B的表达， 补充BRD 4-NUT在维持NC生长中的致癌激活。以该途径为靶点， tazemetostat（taz）与BETi高度协同，并将在所提出的GEM模型中进行探索。 现在认识到，NC具有免疫逃避性肿瘤微环境（TME），并且可以响应于免疫应答。 免疫调节疗法已知表观遗传调节剂如taz和BETi促进抗肿瘤活性。 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 编码BRD 4-NUT的鼠Brd 4与人NUTM 1的他莫昔芬诱导的条件敲入融合物 融合癌蛋白在我们的NC GEM（“mNC”）中形成的侵袭性肿瘤提供了最明确的证据 BRD 4-NUT是这种癌症的唯一驱动力。mNC与人NC非常相似，表现出快速生长， 转移性扩散和难以区分的组织病理学和免疫表型。此外，免疫细胞 mNCTME的组成也类似于人NC的组成。我们将利用我们新颖的创业板来解决 以下目标：1.建立NC GEM（mNC）对人类NC（hNC）生物学的适用性。2.制定 NUT癌的改进的初级治疗。3.探讨预防NC复发的方法。