PROTEOMIC AND GENOMIC PROFILING OF SAMPLES FROM THYROID CANCER PATIENTS

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。列出的机构是 中心，不一定是研究者的机构。 问题陈述： 大约90%的内分泌癌是甲状腺癌（Hundahl等人，1998; Correra等人，1995年）。预计今年将有超过33，500例甲状腺癌（TC）新发病例，预计将有1，530名美国人死于该疾病（Jemal et al.，2007年）。 早期发现可以提高生存率。目前，大多数诊断为甲状腺结节的患者都需要进行细针穿刺（FNA）活检，以确定结节是否为恶性。这种方法有助于确定结节是否由良性滤泡上皮细胞组成。在许多滤泡性肿瘤病例中，由于细胞学特征相似，在FNA标本检查期间，良性肿瘤无法与恶性肿瘤区分。在FNA样本中不可能评估包膜和血管浸润。在这些情况下，手术通常被推荐用于治疗甲状腺结节，从那里获得了可疑的抽吸。事实上，多达80%的未确定病例在进行手术后的组织学检查期间被鉴定为良性肿瘤（Deveci等人，2006年）。 发展快速、敏感、特异、可靠和廉价的方法来鉴别诊断癌症和监测治疗后的缓解是必要的。如果成功，它将减少甚至消除许多FNA结果可疑的患者不必要的甲状腺切除术。 我们认为，在此提出的研究将导致开发这样一种工具。 本研究的目的是利用从血清和血浆以及FNA标本中提取的RNA和蛋白质，探索一组特异于TC类型的蛋白质组学和基因组学生物标志物。这些结果随后将与来自手术期间切除的肿瘤样品的一组蛋白质组学和基因组生物标志物以及相应的术后组织病理学诊断进行比较。 因此，将确定合适的基因组和蛋白质组标志物的较小子集。这将有助于设计一个更大的后续实验，以确定来自具有已知临床结果的患者的甲状腺活检的混合细胞群体的生物标志物谱。专注于较少的蛋白质芯片类型将允许增加来自不同TC类型的复制数量，并获得更具统计学意义的结果。从长远来看，可以开发一种快速TC诊断系统。 拟议研究背后的具体假设是，每种类型的TC都可以通过一组特定的生物标志物来表征。 研究方法： 进行FNA用于诊断目的，而不是专门用于本研究。对于本研究，将在常规FNA程序期间采集额外的抽吸物，用于蛋白质组学研究。 将从同意该程序的患者中收集血清。如果进行手术，则将保存用于组织病理学评价的组织碎片用于本研究。 FNA手术：在有或没有局部麻醉的情况下进行。使用27-25号针头收集约1-6次抽吸。手术后，患者可能会出现轻微不适和皮肤变色或血肿。手术由经验丰富的内分泌学家或病理学家进行。 采血：每例患者将采集约15- 20 ml血液 肿瘤样本：如果进行手术，切除肿瘤的碎片（约250- 500 mg）将被快速冷冻并储存在-80 ℃下用于进一步分析。 样本处理和数据分析：将从FNA和组织样本中提取RNA和蛋白质样本，并储存在-80 ℃下，直至分别使用微阵列芯片SELDI-TOF芯片进行处理。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Statement of the problem: Approximately 90% of all endocrine cancers are thyroid carcinomas (Hundahl et al., 1998; Correra et al., 1995). Over 33,500 new cases of thyroid cancer (TC) are expected this year, and 1,530 Americans are expected to die from the disease (Jemal et al., 2007). Early detection can increase survival rate. Currently, most patients diagnosed with a thyroid nodule are referred for fine needle aspirate (FNA) biopsy to determine if the nodule is malignant or not. This method is helpful in determining if the nodule consists of benign follicular epithelium cells. In many cases of follicular neoplasms, due to similarity in cytologic features, benign tumor cannot be differentiated from malignant cancer during examination of FNA specimen. It is impossible to evaluate capsular and vascular invasion in FNA samples. In these cases, surgery is routinely recommended for the treatment of thyroid nodules from which a suspicious aspiration has been obtained. In fact, as many as 80% of the undetermined cases are identified as benign tumors during histological examinations after the surgery is performed (Deveci et al., 2006). The development of rapid, sensitive, specific, reliable, and inexpensive methods to differentially diagnose cancers and to monitor remission after treatment is necessary. If successful, it would reduce or even eliminate unnecessary thyroidectomies in many patients with suspicious FNA results. We believe that the study proposed here will lead to development of such a tool. The objective of the study is to explore a set of proteomic and genomic biomarkers specific to the type of TC using RNA and proteins extracted from the blood serum and plasma and from FNA specimens. These results would later be compared with a set of proteomic and genomic biomarkers from tumor samples excised during the surgery and with the corresponding post surgical histo-pathological diagnosis. As a result, a smaller subset of suitable genomic and proteomic markers will be determined. This will help design a larger follow-up experiment to identify biomarker profiles of mixed cell populations from thyroid biopsies from patients with known clinical outcomes. Focusing on fewer protein chip types will allow increasing the number of replications from different TC types and obtaining results that will be much more statistically significant. In the long run, a system of rapid TC diagnostics may be developed. The specific hypothesis behind the proposed research is that each type of TC can be characterized by a specific set of biomarkers. Methods: FNAs are preformed for diagnostic purposes and not specifically for this study. For this study, additional aspirates would be collected during the routine FNA procedure to be used in the proteomic study. Blood serum would be collected from the patients consenting to this procedure. If surgery is performed, a fragment of tissue saved for histo-pathological evaluation would be used in this study. FNA procedure: is performed with or without local anesthesia. About 1-6 aspirations are collected using 27-25 gauge needle. After the procedure patient may expect slight discomfort and skin discoloration or hematoma. The procedure is performed by experienced endocrinologist or pathologist. Blood collection: approximately 15-20ml of blood will be collected from each patient Tumor sample: if the surgery is performed, a fragment of the removed tumor (approx. 250-500mg) will be snap frozen and stored in -80C) for further analysis. Sample processing and data analysis: RNA and protein samples will be extracted from FNA and tissue samples and stored in -80C until processed using microarray chips SELDI-TOF chips respectively.