Regulation of Extracellular Matrix 1(ECM1) and its role in thyroid carcinogenesis

应聘者描述(由申请人提供)：应聘者为助理教授(终身制)，主要对内分泌外科肿瘤学和基础科学研究方面的培训感兴趣。她非常致力于一个长期的职业目标，即在甲状腺癌研究领域确立自己的独立调查者地位。她一直在一个非常合作和协同的环境中工作，并对她的学术成功做出了大量的机构承诺。她正在申请这一奖项，目的是最终能够申请NIH R01拨款，并建立自己的研究项目。她对这一奖励机制的职业发展计划包括正式的课程工作和参加与她的研究领域相关的研讨会和会议，目的是增加癌症基因调控机制的教学和实践知识。此外，她将与她的导师和研究审查委员会密切互动，以确保她进步到独立。甲状腺癌是影响内分泌腺的最常见的恶性肿瘤，其发病率在过去30年里一直在上升。这并不是一种致命的恶性肿瘤。然而，据报道，复发疾病(发生在25%的患者中)会增加随后的死亡率，即使是分化良好的甲状腺癌也是如此。分化良好的肿瘤通常采用手术和放射性碘消融治疗。然而，复发往往对放射性碘不太敏感，而且不可能可靠地预测哪些肿瘤会复发或转移。此外，一些类型的甲状腺癌，如间变性肿瘤，侵袭性很强，目前还没有有效的治疗方法。初步研究表明，细胞外基质1(ECM1)在间变性甲状腺癌中过度表达，其表达可能是分化型甲状腺恶性肿瘤的预后指标。使用细胞系模型的其他初步研究表明，Tfap2c可能调节ECM1的表达。这项提议的工作背后的假设是，ECM1是甲状腺癌中一个重要的预后标记物，它增加了转移潜力，其表达受TFAP2调节。这项应用旨在扩大我们对ECM1过度表达的调控及其在预后中的作用的理解，具体目的如下。1)确定ECM1的转录调控机制，特别是Tfap2c；2)检测ECM1的过度表达和沉默对肿瘤细胞侵袭和转移潜能的作用；3)检测ECM1表达作为甲状腺癌患者预后决定因素的作用，并确定它是否与Tfap2c共表达。荧光素酶分析将用于定义ECM1的启动子区域，染色质免疫沉淀将用于确定Tfap2c是否与该区域结合。ECM1过表达的影响将通过体外侵袭和迁移试验以及体内原位小鼠模型进行评估。最后，我们将通过实时RT-PCR和免疫组织化学方法对一组以医院为基础的甲状腺癌患者进行ECM1表达检测，以确定其是否与临床结果相关。这项研究的意义和与健康相关的是，它将增加我们对甲状腺癌侵袭性和预后不良表型的分子和遗传学基础的理解。这将有望导致对它们的发展的新的见解，并增强我们发现这些和其他低风险肿瘤的新的和定向的治疗方法的能力。

DESCRIPTION (provided by applicant): The candidate is an Assistant Professor (tenure track) with primary clinical interest in Endocrine Surgical Oncology and training in basic science research. She is very committed to a long-term career goal of establishing herself as an independent investigator in the area of thyroid cancer research. She has been working in a very collaborative and synergistic environment with substantial institutional commitment to her academic success. She is applying for this award with the intent of eventually being able to apply for an NIH R01 grant and establish her own research program. Her career development plan for this award mechanism includes formal course work and attendance at seminars and conferences pertinent to her field of research with a goal to increase both didactic and practical knowledge in the mechanisms of gene regulation in cancers. In addition, she will interact closely with her mentors and Research Review Committee to ensure that she progresses to independence. Thyroid cancer is the most common malignancy affecting the endocrine glands and its incidence rates have been rising over the past 30 years. It is not a uniformly fatal malignancy. However, recurrent disease (occurring in 25% of patients) has been reported to increase subsequent mortality, even from well- differentiated thyroid cancers. Well-differentiated tumors are typically treated with surgery and radio-active iodine ablation. However, recurrences are often less responsive to radioactive iodine and it is not possible to reliably predict which tumors will recur or metastasize. In addition, some types of thyroid cancer such as anaplastic tumors are very aggressive and currently have no effective therapy. Preliminary studies suggest that Extracellular Matrix 1 (ECM1) is over-expressed in anaplastic thyroid cancers and that its expression may be a prognostic indicator in differentiated thyroid malignancies. Additional preliminary studies using cell line models indicate that TFAP2C likely regulates ECM1 expression. The hypothesis underlying the proposed work is that ECM1 is an important prognostic marker in thyroid cancer that confers increased metastatic potential and that its expression is regulated by TFAP2. This application proposes to expand our understanding of the regulation of ECM1 overexpression and its role in prognosis with the following specific aims. 1) Determine the mechanisms of transcriptional regulation of ECM1, with particular reference to TFAP2C; 2) Examine the role of ECM1 over-expression and silencing on invasive and metastatic potential of tumor cells and 3) Examine the role of ECM1 expression as a determinant of outcome in patients with thyroid cancer and determine whether it is co-expressed with TFAP2C. Luciferase assays will be used to define the promoter region of ECM1 and chromatin immunoprecipitation will be used to determine if TFAP2C binds this region. The effects of ECM1 overexpression will be evaluated using in vitro invasion and migration assays and in vivo in an orthotopic mouse model. Lastly, ECM1 expression will be examined by real-time RT-PCR and immunohistochemistry in a hospital-based cohort of patients with thyroid cancer to determine whether it correlates with clinical outcomes. The significance and health-relatedness of this research endeavor is that it will increase our understanding of the molecular and genetic basis of the aggressive and poor-prognosis phenotype of thyroid cancer. This will hopefully lead to new insights regarding their development and enhance our ability to discover novel and directed therapies for these and other poor-risk tumors.