Pathogenesis and Treatment of NUT-Midline Carcinoma

NUT-中线癌的发病机制和治疗

• 批准号: 8579312
• 负责人: Christopher A French
• 金额: $ 29.26万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-30 至 2018-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8579312
• 关键词: Adenocarcinoma; Binding; Bromodomain; Cancer Etiology; Cancer Model; Carcinoma; Cell Line; Chimeric Proteins; Chromatin; Chromosomal Rearrangement; Code; Collaborations; Combined Modality Therapy; DNA Sequence Rearrangement; Disease; EP300 gene; Epidermal Growth Factor Receptor; Event; Family; Future; Gene Family; Gene Targeting; Genes; Genetic Transcription; Genomics; Goals; Growth; HRAS gene; Health; Histone Deacetylase Inhibitor; Histones; In Vitro; Institution; Karyotype; MYC Family Protein; Maintenance; Malignant Epithelial Cell; Malignant Neoplasms; Malignant neoplasm of lung; Mass Spectrum Analysis; Modeling; Molecular; Mutate; Mutation; Normal Cell; Oncogene Proteins; Oncogenes; Oncogenic; Outcome Study; Parents; Pathogenesis; Pathway Analysis; Pathway interactions; Patients; Pharmaceutical Preparations; Phase I Clinical Trials; Post-Translational Protein Processing; Proteins; RNA; Reading; Regulation; Reporting; Respiratory System; Respiratory tract structure; Role; Small Interfering RNA; Squamous cell carcinoma; TP53 gene; Testing; Transferase; Translations; Tyrosine Kinase Inhibitor; Up-Regulation; Work; base; cancer therapy; cancer type; effective therapy; exome sequencing; field study; genome-wide; in vivo; inhibitor/antagonist; killings; member; mutant; neoplastic cell; novel; protein complex; small molecule; tool; tumor

DESCRIPTION (provided by applicant): The focus of this project is a rare, highly aggressive, and incurable subtype of squamous cell carcinoma, genetically defined by rearrangement of the NUT gene, and termed midline carcinoma (NMC). The goals of the enclosed proposal are 1) to determine the pathogenesis and 2) effective therapy of NMC. NMC is characterized by a simple karyotype harboring a single translocation involving the NUT gene with members of the bromodomain-containing BET family, most commonly BRD3 or BRD4. The resultant BRD- NUT fusion protein is causative in this cancer, acting to block differentiation and maintain NMC cells in a perpetually proliferative state. Several pieces of evidence have led us formulate a model whereby BRD-NUT blocks differentiation by sequestering chromatin "writers" necessary for transcription, such as histone acetyl- transferases (HATs), away from genes required for differentiation to limited genomic regions where the bromodomains of BRD "read" and activate transcription of pro-growth, anti-differentiative genes. Indeed, drug inhibitors of the acetyl-histone binding bromodomains (BETi) or of histone deacetylases unblock the BRD-NUT induced blockade on differentiation, resulting in differentiation and arrested proliferation of NMC cells in vitro and in vivo. Both approaches have been used therapeutically in patients. In support of the above model, recent findings indicate that the transcriptional upregulation of the common cancer gene, MYC, by BRD4-NUT is required and can replace BRD4-NUT's function to block differentiation. The first aim, to identify the molecular mechanisms by which BRD4-NUT leads to the MYC-dependent blockade of differentiation in NMC cells, will be accomplished by isolating and sequencing genes with which BRD4-NUT associates, and correlating with changes that occur in expression of those genes under conditions which promote differentiation in NMC cells. The gene targets of BRD4-NUT identified in these studies will be correlated with findings acquired from the genome-wide siRNA knockdown, which will identify specific genes whose expression is required for the blockade of differentiation in NMC cells. After a list of key target genes is identified and prioritized based on pathway analysis, they will be tested for their effects on MYC protein and RNA levels to identify which BRD4-NUT target genes are required to upregulate MYC levels. In addition, identification of BRD4-NUT-containing protein complexes by mass-spectrometry will be performed to identify proteins which collaborate with BRD4-NUT to upregulate MYC. The second aim, an exploratory aim, to identify mutations in NMC cells that collaborate with BRD4-NUT in the blockade of differentiation and maintenance of growth of NMC cells, will be accomplished by sequencing all coding genes of approximately ten NMC tumors. The purpose of this aim is also to identify mutant proteins which aid BRD4-NUT oncogenic activity that can be targeted by small molecule inhibitors in the treatment of NMC. PHS 398/2590 (Rev. 09/04) Page Continuation Format Page

描述（由申请人提供）：该项目的重点是鳞状细胞癌的罕见，高度侵略性且无法治愈的亚型，是通过坚果基因重排的基因定义的，并称为中线癌（NMC）。封闭建议的目标是1）确定发病机理和2）NMC的有效治疗。 NMC的特征是一种简单的核型，具有单个易位，涉及螺母基因与含溴脱域的BET家族的成员，最常见的是BRD3或BRD4。在该癌症中，由此产生的Brd-Nut融合蛋白是致病性的，可阻止分化并维持永久增殖状态的NMC细胞。几个证据导致我们制定了一个模型，BRD-NUT通过隔离转录所需的染色质“作者”来阻止分化，例如组蛋白乙酰基转移酶（HATS），远离分化为有限的基因组区域所需的基因，其中BRD的BRD“ bromodomain”读取“读取”和激活培训的转录。实际上，丙酮结合溴a（BETI）或组蛋白脱乙酰基酶的药物抑制剂解开BRD-NUT引起的分化后阻断，从而导致NMC的分化和停滞 细胞体外和体内。两种方法都在患者中使用治疗。 为了支持上述模型，最新发现表明，需要BRD4-NUT对公共癌基因MYC的转录上调，并且可以替代BRD4-NUT的功能以阻止分化。 第一个目的是通过隔离和测序BRD4-NUT辅助物来确定BRD4-NUT导致NMC细胞中MYC依赖性分化的封锁的分子机制，并与在NMC细胞中促进分化的这些基因表达中发生的变化相关。这些研究中鉴定出的BRD4-NUT的基因靶标将与从全基因组siRNA敲低的发现相关，该发现将确定其表达的特定基因，其表达是NMC细胞中分化的封锁所必需的。根据途径分析确定关键靶基因列表并确定优先级后，将对其进行测试 对MYC蛋白和RNA水平的影响，以确定需要哪些BRD4-NUT靶基因上调MYC水平。此外，将通过质量光谱法对含BRD4-NUT的蛋白质复合物进行鉴定，以识别与BRD4-NUT合作以上调MYC的蛋白质。 第二个目的是探索性的目的，是通过对大约十个NMC肿瘤的所有编码基因进行测序来实现NMC细胞中与BRD4-NUT合作的NMC细胞中与BRD4-NUT合作的突变。该目的的目的还是鉴定突变蛋白，这些突变蛋白有助于BRD4-NUT致癌活性，而小分子抑制剂可以靶向NMC。 PHS 398/2590（修订版09/04）页面延续格式页面