Oral Carcinogenesis

口腔癌发生

• 批准号: 8344120
• 负责人: J Gutkind
• 金额: $ 173.95万
• 依托单位: NATIONAL INSTITUTE OF DENTAL & CRANIOFACIAL RESEARCH
• 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/8344120
• 关键词: Address; Affect; Alcohol consumption; Animal Model; Animals; Apoptotic; Area; Biological; Biological Markers; Biological Models; Cancer Detection; Carbon Nanotubes; Cell Differentiation process; Cell Surface Receptors; Cells; Cervical lymph node group; Cessation of life; Chemopreventive Agent; Clinical; Collaborations; Communities; DNA; Detection; Development; Diagnostic; Disease Progression; Drug Delivery Systems; Epidermal Growth Factor Receptor; Epithelial; Evaluation; Event; Evolution; Exhibits; Extramural Activities; Future; Gene Expression; Gene Silencing; Genes; Genetic; Genetically Engineered Mouse; Genomics; Glutathione; Goals; Gold; Growth Factor; Head and Neck Squamous Cell Carcinoma; Human; Institution; Integrins; Interleukin-6; Interleukin-8; Invaded; Investigation; Knowledge; Label; Laboratories; Lesion; Lip structure; Lymph Node Involvement; Lymphangiogenesis; Lymphatic vessel; Malignant Neoplasms; Memorial Sloan-Kettering Cancer Center; Microfluidics; Micrometastasis; Microscopy; Modality; Modeling; Molecular; Molecular Profiling; Molecular Target; Monitor; Mouth Neoplasms; Mus; Mutation; Nanotubes; Nature; Neoplasm Metastasis; Oncogenic; Oral; Oral Tobacco; Oral cavity; Oral mucous membrane structure; Outcome; PTGS2 gene; Pathway interactions; Patients; Pattern; Pharyngeal structure; Population; Positioning Attribute; Premalignant; Prevention; Prevention strategy; Primary Neoplasm; Prognostic Factor; Prostaglandin Receptor; Protein-Serine-Threonine Kinases; Proteins; Proteomics; RNA; Reporting; Reproducibility; Research; Risk Factors; Sampling; Screening for cancer; Sentinel Lymph Node; Series; Serum; Signal Transduction; Sirolimus; Specimen; Stem cells; Subfamily lentivirinae; Surface; System; Technology; Testing; Tetracyclines; Therapeutic; Tobacco use; Tongue; Treatment Effectiveness; Tumor Suppressor Genes; Vascular Endothelial Growth Factor C; analog; base; cancer cell; cancer genomics; cancer therapy; cell growth; forest; human FRAP1 protein; implantation; improved; interest; laser capture microdissection; lymph nodes; mTOR Inhibitor; mTOR inhibition; malignant mouth neoplasm; mouse model; nanoparticle; neoplastic; notch protein; novel; novel strategies; novel therapeutic intervention; oral carcinogenesis; particle; prevent; prognostic; programs; protein expression; receptor; recombinase; research clinical testing; sensor; single walled carbon nanotube; sphingosine kinase; therapeutic target; tumor; tumor xenograft; tumorigenesis; two-photon

30% Effort. Genomic and proteomic approaches to understand oral cancer Although risk factors for HNSCC, such as alcohol and tobacco consumption, are well recognized, the molecular mechanisms responsible for this malignancy are still not fully understood. We have used a number of novel approaches to investigate gene and protein expression profiles in HNSCC. We have shown that laser capture microdissection (LCM) can be used to procure specific cell populations from heterogenous tumor samples, and that LCM-procured material can be used effectively to extract RNA, DNA, and proteins. We have teamed up with other research institutions to conduct gene and protein expression analysis of HNSCC by combining LCM, gene arrays, and proteomic platforms. These efforts have already provided a wealth of information about the distinctive pattern of gene and protein expression in HNSCC. Gene and protein expression analysis: : In prior gene array analysis efforts, we have identified numerous genes that were differentially expressed in normal oral mucosa and cancer, and many distinct genes when comparing cancers associated with betel quid chewing and tobacco use. This body of information enabled us to identify a large number of molecules whose contributions to HNSCC progression are now under investigation in our laboratory and in the extramural community. Examples of molecules that were evaluated during this reporting period include sphingosine kinase, distinct integrins, and COX-2 and prostaglandin receptors. In addition, these studies also revealed that the expression of VEGF-C, a potent lymphangiogenic growth factor, is a shared feature of the most metastatic HNSCC lesions, which provided the rationale for current efforts investigating whether interfering with VEGF-C can halt the metastatic spread of HNSCC. Carbon nanotubes: immunosensensors for cancer biomarkers and drug delivery systems for cell-surface receptor-guided cancer therapy: The genomic and proteomic analysis of HNSCC may now allow the development of novel biomarkers of diagnostic and prognostic value. We continued teaming up with J. Rusling at UCONN to develop nanotube-based systems for the detection of premalignant lesions and micrometastasis in sentinel lymph nodes.. In particular, building on our prior studies using single-wall carbon nanotube (SWNT) forest platforms, we have now developed an ultrasensitive immunosensor based on a glutathione-protected gold nanoparticle (GSH-AuNP) sensor surface. When combined with novel massively labeled paramagnetic particles for the electrochemical detection of the cancer biomarker interleukin 8 (IL-8), we obtained an unprecedented detection limit in the attomolar range, with high level of reproducibility and accuracy. This nanoparticle-based strategy for single-protein sensors has great promise for the future development of nanodetection arrays for clinical cancer screening and therapy monitoring. Indeed, we have now initiated an effort to extend these technologies to an array platform, and succeeded in developing a microfluidic electrochemical immunoarray for ultrasensitive detection of two cancer biomarkers IL-8 and IL-6, in human serum samples. 40% Effort.Dysregulated signaling networks in HNSCC: novel mechanism-based approaches for HSNCC prevention and treatment There is an urgent need for new treatment options for HNSCC patients, considering that their overall 5-year survival is relatively low (50%) and has not improved much over the past 3 decades. The emerging information on the nature of the deregulated molecular mechanisms responsible for HNSCC progression has provided the possibility of exploring new mechanisms-based therapeutic approaches for HNSCC. For example, we have observed that persistent activation of the serine-threonine kinases mTOR is frequent event in HNSCC, and that inhibition of mTOR by the use of rapamycin causes the rapid decrease in the level of pS6 (a downstream target of mTOR) and the apoptotic death of HNSCC tumor xenografts, thereby causing tumor regression. These efforts have identified the Akt-mTOR pathway as a potential therapeutic target for HNSCC. Inhibition of mTOR by rapamycin prevents lymphangiogenesis and locoregional lymph node metastasis in a new HNSCC orthotopic model. HNSCCs often metastasize to locoregional lymph nodes, and lymph node involvement represents the most important prognostic factor of poor clinical outcome. Of interest, in collaboration with Bhuvanesh Singh, MSKCC, we observed that the activation of mTOR represents a widespread event in human clinical specimens of HNSCC invading locoregional lymph nodes. Furthermore, both primary and metastatic experimental HNSCC lesions exhibited elevated mTOR activity. To begin exploring the contribution of mTOR to HNSCC metastasis we developed an orthotopic model consisting in the implantation of fluorescently labeled HNSCC cells into the tongues of immunecompromised mice. These orthotopic tumors spontaneously metastasize to the cervical lymph nodes, where the presence of HNSCC cells can be revealed by histological evaluation. Together with Roberto Weigert, we visualized the tumoral cells invading the tongue and within the lymphatic vessels using intravital two-photon microscopy. The ability to monitor and quantitate lymph node invasion in this model system enabled us to explore whether the blockade of mTOR could impact on HNSCC metastasis. We found that inhibition of mTOR with rapamycin diminished lymphangiogenesis in the primary tumors and prevented the dissemination of HNSCC cancer cells to the cervical lymph nodes, thereby prolonging animal survival. These findings may provide a rationale for the future clinical evaluation of mTOR inhibitors, including rapamycin and its analogs, as part of a molecular-targeted metastasis preventive strategy for the treatment of HNSCC patients. 30% Effort. Animal models for oral malignancies A major limitation in the area of HNSCC research is the limited availability of animal models to test the validity of current genetic paradigms of tumorigenesis, and to explore the effectiveness of treatment modalities or chemopreventive approaches. Novel genetically-defined and chemically induced oral-specific animal models to study SCC: We have recently made significant contributions to the development of genetically engineered mouse models (GEMM) for HNSCC. We have continued with these studies, including the recent analysis of the interplay between the TGF-β and Akt-mTOR pathway in collaboration with Ashok Kulkarni. In this line of research, we have now focused our efforts in this area in the development of oral specific systems enabling the activation/inactivation of genes in the epithelial stem cell compartment. Current studies are aimed at recapitulating HNSCC progression, thus providing a suitable system to investigate targeted approaches to halt tumor development. In addition to the use of Cre-recombinase and tetracycline-inducible systems to delete relevant tumor suppressor gene products (i.e., p16, Notch, Pten, p53, TGF-β receptors) and to express oncogenic molecules (i.e., Ras, EGFR), we are now making a new effort in two different directions. We are attempting to develop a high throughput animal model by the use of lentiviruses that are only active in the epithelial stem cell compartment. This approach may position us to address the biological relevance of newly identified genetic alterations that are expected to result from the oncogenomic studies in our intra-and extra-mural community.

30% 的努力。了解口腔癌的基因组学和蛋白质组学方法 尽管 HNSCC 的危险因素（例如饮酒和吸烟）已得到广泛认可，但导致这种恶性肿瘤的分子机制仍不完全清楚。我们使用了许多新方法来研究 HNSCC 中的基因和蛋白质表达谱。我们已经证明，激光捕获显微切割 (LCM) 可用于从异质肿瘤样本中获取特定细胞群，并且 LCM 获取的材料可有效用于提取 RNA、DNA 和蛋白质。我们与其他研究机构合作，结合LCM、基因芯片和蛋白质组平台进行HNSCC的基因和蛋白表达分析。这些努力已经提供了有关 HNSCC 中基因和蛋白质表达的独特模式的大量信息。 基因和蛋白质表达分析：：在之前的基因阵列分析工作中，我们已经鉴定出许多在正常口腔粘膜和癌症中差异表达的基因，以及在比较与咀嚼槟榔和吸烟相关的癌症时许多不同的基因。这些信息使我们能够识别出大量对 HNSCC 进展有贡献的分子，目前我们的实验室和校外社区正在研究这些分子。本报告期间评估的分子示例包括鞘氨醇激酶、不同的整合素以及 COX-2 和前列腺素受体。此外，这些研究还表明，VEGF-C（一种有效的淋巴管生成因子）的表达是大多数转移性 HNSCC 病变的共同特征，这为当前研究干扰 VEGF-C 是否可以阻止转移性 HNSCC 病变的研究提供了理论基础。 HNSCC 的传播。 碳纳米管：用于癌症生物标志物的免疫传感器和用于细胞表面受体引导的癌症治疗的药物输送系统：HNSCC 的基因组和蛋白质组分析现在可以允许开发具有诊断和预后价值的新型生物标志物。我们继续与 UCONN 的 J. Rusling 合作开发基于纳米管的系统，用于检测前哨淋巴结的癌前病变和微转移。特别是，在我们之前使用单壁碳纳米管 (SWNT) 森林平台的研究的基础上，我们现在开发了一种基于谷胱甘肽保护的金纳米颗粒 (GSH-AuNP) 传感器表面的超灵敏免疫传感器。当与新型大规模标记顺磁颗粒结合用于癌症生物标志物白细胞介素 8 (IL-8) 的电化学检测时，我们在阿摩尔范围内获得了前所未有的检测限，并且具有高水平的重现性和准确性。这种基于纳米颗粒的单蛋白传感器策略对于临床癌症筛查和治疗监测的纳米检测阵列的未来发展具有巨大的前景。事实上，我们现在已经开始努力将这些技术扩展到阵列平台，并成功开发了微流控电化学免疫阵列，用于超灵敏检测人血清样本中的两种癌症生物标志物 IL-8 和 IL-6。 40% Effort.HNSCC 信号网络失调：基于机制的 HSNCC 预防和治疗新方法 考虑到 HNSCC 患者的总体 5 年生存率相对较低（50%），并且在过去 30 年来没有太大改善，因此迫切需要新的治疗方案。 关于导致 HNSCC 进展的失调分子机制性质的新信息为探索新的基于机制的 HNSCC 治疗方法提供了可能性。例如，我们观察到丝氨酸-苏氨酸激酶 mTOR 的持续激活是 HNSCC 中的常见事件，并且使用雷帕霉素抑制 mTOR 会导致 pS6（mTOR 的下游靶标）和HNSCC 肿瘤异种移植物凋亡，从而导致肿瘤消退。这些努力已将 Akt-mTOR 通路确定为 HNSCC 的潜在治疗靶点。 在新的 HNSCC 原位模型中，雷帕霉素抑制 mTOR 可预防淋巴管生成和局部淋巴结转移。 HNSCC 经常转移至局部淋巴结，淋巴结受累是临床预后不良的最重要的预后因素。有趣的是，我们与 MSKCC 的 Bhuvanesh Singh 合作，观察到 mTOR 的激活代表了侵入局部淋巴结的 HNSCC 人类临床标本中的广泛事件。此外，原发性和转移性实验性 HNSCC 病变均表现出 mTOR 活性升高。为了开始探索 mTOR 对 HNSCC 转移的贡献，我们开发了一种原位模型，即将荧光标记的 HNSCC 细胞植入免疫受损小鼠的舌头中。这些原位肿瘤自发转移到颈部淋巴结，通过组织学评估可以揭示颈部淋巴结中是否存在 HNSCC 细胞。我们与 Roberto Weigert 一起，使用活体双光子显微镜观察了侵入舌头和淋巴管内的肿瘤细胞。在该模型系统中监测和定量淋巴结侵袭的能力使我们能够探索 mTOR 的阻断是否会影响 HNSCC 转移。我们发现，用雷帕霉素抑制 mTOR 可以减少原发肿瘤中的淋巴管生成，并阻止 HNSCC 癌细胞扩散到颈部淋巴结，从而延长动物的生存期。这些发现可能为 mTOR 抑制剂（包括雷帕霉素及其类似物）的未来临床评估提供依据，作为治疗 HNSCC 患者的分子靶向转移预防策略的一部分。 30% 的努力。口腔恶性肿瘤动物模型 HNSCC 研究领域的一个主要限制是动物模型的可用性有限，无法测试当前肿瘤发生遗传范式的有效性，并探索治疗方式或化学预防方法的有效性。 用于研究 SCC 的新型基因定义和化学诱导的口腔特异性动物模型：我们最近为 HNSCC 基因工程小鼠模型 (GEMM) 的开发做出了重大贡献。我们继续进行这些研究，包括最近与 Ashok Kulkarni 合作分析 TGF-β 和 Akt-mTOR 通路之间的相互作用。在这一领域的研究中，我们现在将工作重点放在开发口腔特异性系统上，以激活/灭活上皮干细胞区室中的基因。目前的研究旨在重现 HNSCC 的进展，从而提供一个合适的系统来研究阻止肿瘤发展的靶向方法。除了使用Cre重组酶和四环素诱导系统删除相关抑癌基因产物（即p16、Notch、Pten、p53、TGF-β受体）并表达致癌分子（即Ras、EGFR）之外，我们现在正在两个不同的方向做出新的努力。我们正在尝试使用仅在上皮干细胞区室中活跃的慢病毒来开发高通量动物模型。这种方法可能使我们能够解决新发现的遗传改变的生物学相关性，这些改变预计是由我们校内和校外社区的肿瘤基因组研究产生的。