Oral Carcinogenesis

口腔癌发生

• 批准号: 8148625
• 负责人: J Gutkind
• 金额: $ 160.85万
• 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/8148625
• 关键词: Achievement; Address; Affect; Alcohols; Animal Model; Animals; Antineoplastic Agents; Apoptotic; Area; Attention; Biological; Biological Models; Carbon Nanotubes; Cell Surface Receptors; Cells; Cervical lymph node group; Cessation of life; Chemopreventive Agent; Cisplatin; Clinical; Collaborations; Communities; DNA; Detection; Development; Diagnostic; Disease Progression; Drug Delivery Systems; EGF gene; EPS8 gene; Epidermal Growth Factor Receptor; Epithelial; Evaluation; Event; Evolution; Exhibits; Extramural Activities; Future; Gene Proteins; Gene Silencing; Genes; Genetic; Genetically Engineered Mouse; Genomics; Goals; Growth Factor; Head and Neck Squamous Cell Carcinoma; Human; Image; Institution; Invaded; Investigation; Knowledge; Label; Laboratories; Lesion; Lip structure; Lymph Node Involvement; Lymphangiogenesis; Lymphatic vessel; Malignant Neoplasms; Memorial Sloan-Kettering Cancer Center; Micrometastasis; Microscopy; Modality; Modeling; Molecular; Molecular Profiling; Molecular Target; Monitor; Mouth Neoplasms; Mus; Mutation; Nature; Neoplasm Metastasis; Oral; Oral Tobacco; Oral cavity; Oral mucous membrane structure; PTGS2 gene; Pathway interactions; Patients; Pattern; Pharmaceutical Preparations; Pharyngeal structure; Platinum; Positioning Attribute; Premalignant; Prevention; Prevention strategy; Primary Neoplasm; Prognostic Factor; Prostaglandin Receptor; Protein-Serine-Threonine Kinases; Proteins; Proteomics; Reporting; Research; Risk Factors; Sampling; Sentinel Lymph Node; Series; Serotyping; Serum; Signal Transduction; Sirolimus; Specimen; Stem cells; Subfamily lentivirinae; Surface; System; TP53 gene; Testing; Tetracyclines; Therapeutic; Tissues; Tobacco use; Tongue; Treatment Effectiveness; Tumor Suppressor Genes; Vascular Endothelial Growth Factor C; analog; base; cancer cell; cancer therapy; cell growth; chemotherapeutic agent; forest; human FRAP1 protein; improved; interest; killings; laser capture microdissection; lymph nodes; mTOR Inhibitor; malignant mouth neoplasm; nanomedicine; neoplastic; novel; oral carcinogenesis; prevent; prognostic; programs; protein expression; receptor; recombinase; research clinical testing; single molecule; sphingosine kinase; tumorigenesis

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 between 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, COX-2 and prostaglandin receptors, EPS8, and FOXM1. 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. We succeeded in using single-wall carbon nanotube (SWNT) forest platforms for immunosensing biomarkers in serum and tissue lysates, and for the targeted killing of HNSCC cells, taking advantage of the ease of functionalization of SWNTs. As most HNSCC overexpress EGFR, we attached cisplatin, a first line chemotherapeutic agent, and EGF (for targeting) to SWNTs. EGF-SWNTs were internalized by HNSCC cells depending on the surface overexpression of EGFR. EGF-SWNT-cisplatin bioconjugates caused the rapid death of HNSCC cells, but sparing control cells showing low levels of EGFR. These studies provided the first evidence of targeted killing of cancer cells by guiding SWNTs loaded with an anticancer drug to cancerous cells based on cell surface receptors. We followed this study by performing the first single-molecule imaging of platinum-based drug attached to single-wall carbon nanotubes, a collaborative achievement that has recently received the broad attention of the emerging nanomedicine community. 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, 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. For this line of research, we are also exploring the use of new AAV serotypes based on our ongoing collaborations with Jay Chiorini.

30％的努力。理解口腔癌的基因组和蛋白质组学方法 尽管众所周知，HNSCC的危险因素，例如酒精和烟草消费，但仍未完全了解造成这种恶性肿瘤的分子机制。我们已经使用了许多新型方法来研究HNSCC中的基因和蛋白质表达谱。我们已经表明，激光捕获显微解剖（LCM）可用于从异源肿瘤样品中采购特定的细胞群，并且可以有效地使用LCM所致的材料来提取RNA，DNA和蛋白质。我们已经与其他研究机构合作，通过组合LCM，基因阵列和蛋白质组学平台对HNSCC进行基因和蛋白质表达分析。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：在先前的基因阵列分析工作中，我们已经确定了许多基因在正常口服粘膜和癌症之间差异表达，并且在比较与槟榔咀嚼和烟草使用相关的癌症时，许多不同的基因。这一信息使我们能够确定大量的分子，这些分子对HNSCC进展的贡献现已在我们的实验室和校外社区中进行研究。在此报告期间评估的分子的实例包括鞘氨醇激酶，COX-2和前列腺素受体，EPS8和FOXM1。这些研究还表明，VEGF-C的表达是一种有效的淋巴管生长生长因子，是最转移性HNSCC病变的共同特征，这为当前努力提供了研究的基本原理，研究研究了VEGF-C是否可以停止HNSCC的转移性扩散。 碳纳米管：用于细胞表面受体引导的癌症治疗的癌症生物标志物和药物输送系统的免疫传感器：HNSCC的基因组和蛋白质组学分析现在可以允许发展新型诊断和预后价值的生物标志物。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中的预触发病变和微量表。我们成功地使用了单壁碳纳米管（SWNT）森林平台，以利用SWNT的功能化，从而在血清和组织裂解物中对生物标志物进行免疫传感。作为大多数HNSCC过表达的EGFR，我们将顺铂（第一线化学治疗剂）和EGF（用于靶向）附加到SWNT上。 EGF-SWNT由HNSCC细胞内化，具体取决于EGFR的表面过表达。 EGF-SWNT-CISPLATIN BIOCONJUGATES引起了HNSCC细胞的快速死亡，但较低的控制细胞显示出低水平的EGFR。这些研究提供了基于细胞表面受体的癌细胞中载有抗癌药物的抗癌药物的SWNT，提供了靶向杀死癌细胞的第一个证据。我们遵循了这项研究，通过单壁碳纳米管进行了第一个基于铂的药物的单分子成像，这是一项合作成就，最近受到了新兴纳米医学界的广泛关注。 40％的努力。在HNSCC中进行调节的信号网络：基于HSNCC预防和治疗的新型机制方法 考虑到他们的总体5年生存率相对较低（50％），并且在过去3年中没有太大改善，因此迫切需要针对HNSCC患者的新治疗选择。 有关负责HNSCC进展的放松管制分子机制的性质的新兴信息提供了探索基于HNSCC的新机制治疗方法的可能性。 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.这些努力将AKT-MTOR途径确定为HNSCC的潜在治疗靶点。 雷帕霉素对MTOR抑制可防止新的HNSCC原位模型中的淋巴管生成和局部淋巴结转移。 HNSCCS经常转移到局部淋巴结，淋巴结受累是临床结果不良的最重要的预后因素。有趣的是，与MSKCC的Bhuvanesh Singh合作，我们观察到MTOR的激活代表了HNSCC临床标本的广泛事件，入侵了局部淋巴结。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，该模型包括将荧光标记的HNSCC细胞植入到免疫功能的小鼠的舌头中。这些原位肿瘤自发地转移至宫颈淋巴结，其中HNSCC细胞的存在可以通过组织学评估来揭示。我们与罗伯托·韦格特（Roberto Weigert）一起，使用静脉内两光子显微镜观察了入侵舌头和淋巴管内的肿瘤细胞。在此模型系统中监测和定量淋巴结入侵的能力使我们能够探索MTOR的阻断是否可能影响HNSCC转移。我们发现，用雷帕霉素抑制MTOR在原发性肿瘤中降低了淋巴管生成，并防止了HNSCC癌细胞向宫颈淋巴结传播，从而延长了动物的生存。这些发现可能为包括雷帕霉素及其类似物在内的MTOR抑制剂的未来临床评估提供了理由，这是分子靶向HNSCC患者治疗的分子转移预防策略的一部分。 30％的努力。口腔恶性肿瘤的动物模型 HNSCC研究领域的一个主要局限性是动物模型的可用性有限，以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。 新型的遗传定义和化学诱导的口腔特异性动物模型研究SCC：我们最近为HNSCC的基因工程小鼠模型（GEMM）做出了重要贡献。我们继续进行这些研究，包括与Ashok Kulkarni合作的TGF-β和Akt-Mtor途径之间的相互作用的最新分析。在这一研究中，我们现在将精力集中在这一领域，以开发口服特定系统，从而实现上皮干细胞室中基因的激活/失活。当前的研究旨在概括HNSCC的进展，从而提供了一个合适的系统来研究靶向肿瘤发育的靶向方法。除了使用CRE聚集酶和四环素诱导系统来删除相关的肿瘤抑制基因产物（即P16，PTEN，PTEN，P53，TGF-β受体）和表达致癌分子（即RAS，EGFR），我们现在在两个不同的方面做出了新的努力。我们试图通过使用仅在上皮干细胞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鉴定的遗传改变的生物学相关性，这些遗传改变有望由我们的内部和壁外社区中的致癌基因组研究产生。对于这一研究，我们还根据与Jay Chiorini的持续合作来探讨新的AAV血清型的使用。