Genomics and Proteomics of Head and Neck Cancer

头颈癌的基因组学和蛋白质组学

• 批准号: 7967002
• 负责人: CARTER VAN WAES
• 金额: $ 59.6万
• 依托单位: NATIONAL INSTITUTE ON DEAFNESS AND OTHER COMMUNICATION DISORDERS
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7967002
• 关键词: Adverse effects; Affect; Alcohols; Apoptosis; Apoptotic; Attenuated; Binding; Bioinformatics; Biological Assay; Biological Markers; Biology; Biotechnology; Cancer cell line; Cell Nucleus; Clinical Trials; Cluster Analysis; Collaborations; Cytokine Gene; Deglutition; Development; Diagnosis; Epidermal Growth Factor Receptor; Epithelial; Event; Exposure to; Gefitinib; Gene Cluster; Gene Expression; Gene Expression Profiling; Gene Proteins; Gene Targeting; Genes; Genetic; Genetic Transcription; Genome; Genomics; Goals; Growth; Head and Neck Cancer; Head and Neck Squamous Cell Carcinoma; Human; Human Papillomavirus; In Situ Hybridization; Inflammatory; Laboratories; Larynx; Link; MAP Kinase Gene; Malignant - descriptor; Malignant Neoplasms; Manuscripts; Modeling; Mus; Mutation; NF-kappa B; Neoplasm Metastasis; Normal tissue morphology; Oral cavity; Paclitaxel; Pathogenesis; Pathology; Pathway interactions; Patients; Pharyngeal structure; Phenotype; Phosphoproteins; Play; Preparation; Prevalence; Prevention; Prevention therapy; Preventive Intervention; Protein Microchips; Proteins; Proteomics; Proto-Oncogene Proteins c-akt; Quality of life; Radiation; Radiosurgery; Recurrent Malignant Neoplasm; Regulation; Regulatory Element; Regulon; Reporting; Resistance; Reverse Transcription; Role; STAT3 gene; Signal Pathway; Signal Transduction; Site; Small Interfering RNA; Specimen; Speech; Systems Biology; TNF gene; TP53 gene; Technology; Therapeutic; Tobacco; Transforming Growth Factor beta; Tumor Suppressor Genes; Voice; Yang; activating transcription factor; cDNA Arrays; carcinogenesis; chemotherapy; cytokine; inhibitor/antagonist; malignant phenotype; member; novel; novel strategies; programs; promoter; protein expression; receptor; response; response to injury; small molecule; therapy resistant; transcription factor

We previously showed using microarray profiling biotechnology that stepwise transformation and metastatic progression of SCC in a murine model results in the expression of gene programs related to the signal transcription factor NF-kappaB. Inhibition of NF-kappaB modulated over half the up or down-regulated genes differentially expressed, and attenuated the malignant phenotype, indicating it may be a critical target for prevention or therapy of head and neck cancer. Gene expression profiling and bioinformatic analysis of the promoters of gene clusters differentially expressed in human HNSCC provided evidence for increased prevalence of binding motifs for NF-kappaB as well as other signal transcription factors, such as p53, AP-1, STAT3 and EGR-1 (Yan et al, Genome Biology, 2007). NF-kappaB, p53, AP-1, STAT3 and EGR-1 activation has previously been associated with pathogenesis and therapeutic resistance, and the subsets expressing wt or mt 53 have been reported to differ in response to chemotherapy. These observations suggested the hypothesis that key alterations in a network of signal transcription factors can interact in determining gene expression and development of HNSCC of differing malignant potential and sensitivity or resistance to therapy. Next, we took a systems biology approach to define NF-kappaB regulons, interacting signal pathways and networks in the malignant phenotype of head and neck cancer cell lines differing in p53 status(Yan, Genome Biology, 2008). Unique gene signatures expressed by human HNSCC lines were identified by cDNA microarray, principal components, and cluster analyses and validated by quantitative reverse transcription-PCR (RT-PCR) and in situ hybridization. An inverse relationship between activation of NF-kB, p53 mutation, and inactivation of TGFbeta pathway and target genes suggested the hypothesis that activation of NF-kB may be linked to alteration of p53 and TGFbeta signaling. We discovered that mt p53 represses TGFbeta receptor II expression, attenuating TGF beta signaling, enhancing inflammatory cytokine responsiveness and activation of NF-kB in HNSCC (Cohen et al, Cancer Res 2009). Another relationship identified was between NF-kB member c-REL and p53 members p53, p63 and p73. This led to demonstration that cytokine TNF induced cREL interacts with p63, displacing p73 from growth arrest and apoptotic genes and the nucleus of HNSCC (Lu et al, manuscript submitted). siRNA knockdown of p63 revealed that it down regulates p53 regulated tumor suppressor genes and upregulated numerous NF-kB related genes, through interaction with c-REL. Novel p63, cREL as well as classical p53 and NF-kB sites were defined in these genes and validated by ChIP assay (Yang et al, manuscript in preparation). ChIP sequencing of global gene expression regulated by cREL, p63 and p73 in HNSCC is underway in collaboration with Laboratory of Genetics, NCI. In collaboration with the NCI and Department of Pathology, new proteomic technology, using protein microarrays, was used to analyze EGFR, NF-kB, MAPK, AKT and STAT3 signal phosphoprotein inhibition, and markers of apoptosis in protein extracts from HNSCC lines and specimens from patients in a clinical trial with small molecule inhibitor gefitinib, taxol and radiation.

我们之前使用微阵列分析生物技术证明，小鼠模型中鳞状细胞癌的逐步转化和转移进展会导致与信号转录因子 NF-kappaB 相关的基因程序的表达。抑制 NF-κB 可以调节一半以上差异表达的上调或下调基因，并减弱恶性表型，表明它可能是预防或治疗头颈癌的关键靶点。对人 HNSCC 中差异表达的基因簇启动子的基因表达谱和生物信息学分析提供了 NF-kappaB 以及其他信号转录因子（例如 p53、AP-1、STAT3 和 EGR-1）结合基序普遍性增加的证据（Yan 等人，Genome Biology，2007）。 NF-kappaB、p53、AP-1、STAT3 和 EGR-1 激活先前已被认为与发病机制和治疗耐药相关，并且据报道表达 wt 或 mt 53 的亚群对化疗的反应有所不同。这些观察结果提出了这样的假设：信号转录因子网络中的关键改变可以相互作用，决定不同恶性潜能和对治疗的敏感性或耐药性的 HNSCC 的基因表达和发展。 接下来，我们采用系统生物学方法来定义 NF-kappaB 调节子、p53 状态不同的头颈癌细胞系恶性表型中相互作用的信号通路和网络（Yan，Genome Biology，2008）。通过 cDNA 微阵列、主成分和聚类分析鉴定了人类 HNSCC 系表达的独特基因特征，并通过定量逆转录 PCR (RT-PCR) 和原位杂交进行验证。 NF-kB 的激活、p53 突变以及 TGFbeta 途径和靶基因的失活之间的负相关关系表明 NF-kB 的激活可能与 p53 和 TGFbeta 信号传导的改变有关的假设。我们发现 mt p53 抑制 TGFbeta 受体 II 表达，减弱 TGFbeta 信号传导，增强炎症细胞因子反应性和 HNSCC 中 NF-kB 的激活（Cohen 等人，Cancer Res 2009）。 确定的另一个关系是 NF-kB 成员 c-REL 和 p53 成员 p53、p63 和 p73 之间的关系。这证明细胞因子 TNF 诱导的 cREL 与 p63 相互作用，将 p73 从生长停滞和凋亡基因以及 HNSCC 细胞核中取代（Lu 等人，已提交手稿）。 p63 的 siRNA 敲除表明，它通过与 c-REL 相互作用下调 p53 调节的肿瘤抑制基因并上调许多 NF-kB 相关基因。在这些基因中定义了新型 p63、cREL 以及经典 p53 和 NF-kB 位点，并通过 ChIP 测定进行验证（Yang 等人，手稿正在准备中）。与 NCI 遗传学实验室合作，正在对 HNSCC 中受 cREL、p63 和 p73 调节的全局基因表达进行 ChIP 测序。 与 NCI 和病理学系合作，利用蛋白质微阵列的新蛋白质组学技术，在一项使用小分子抑制剂吉非替尼、紫杉醇和放射治疗的临床试验中，分析了 HNSCC 系和患者样本的蛋白质提取物中的 EGFR、NF-kB、MAPK、AKT 和 STAT3 信号磷蛋白抑制以及细胞凋亡标记物。