PROTEOMIC AND GENOMIC PROFILING OF SAMPLES FROM THYROID CANCER PATIENTS

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 问题陈述： 大约90%的内分泌癌是甲状腺癌(Hundahl等人，1998年；Corera等人，1995年)。预计今年将新增33,500例甲状腺癌(TC)病例，预计将有1,530名美国人死于这种疾病(Jemal等人，2007年)。早期发现可提高存活率。目前，大多数被诊断为甲状腺结节的患者都被转诊为细针抽吸(FNA)活检，以确定结节是否恶性。这种方法有助于确定结节是否由良性滤泡上皮细胞组成。在许多滤泡性肿瘤中，由于细胞学特征相似，在FNA标本检查中无法区分良、恶性肿瘤。在FNA样本中评估包膜和血管侵犯是不可能的。在这些病例中，对于可疑吸入物的甲状腺结节，通常建议手术治疗。事实上，在手术后的组织学检查中，多达80%的未确定病例被确认为良性肿瘤(Deveci等人，2006年)。 开发快速、灵敏、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。我们相信，这里提出的研究将导致开发这样一个工具。 本研究的目的是利用从血清、血浆和FNA标本中提取的RNA和蛋白质，探索一套针对TC类型的蛋白质组和基因组生物标记物。这些结果稍后将与手术中切除的肿瘤样本中的一组蛋白质组和基因组生物标记物以及相应的手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记的较小子集。这将有助于设计一个更大的后续实验，从已知临床结果的患者的甲状腺活检中识别混合细胞群体的生物标记物图谱。专注于更少的蛋白质芯片类型将允许增加不同TC类型的重复数量，并获得更具统计学意义的结果。从长远来看，可以开发一种快速诊断TC的系统。 这项研究背后的具体假设是，每种类型的TC都可以由一组特定的生物标志物来表征。 方法： FNAS是为诊断目的而进行的，而不是专门用于本研究。在这项研究中，额外的吸液将在常规的FNA程序中收集，用于蛋白质组学研究。从同意这一手术的患者身上采集血清。如果进行手术，本研究将使用保存下来用于组织病理学评估的组织片段。 FNA手术：在有或没有局部麻醉的情况下进行。大约1-6个心愿是用27-25号针收集的。手术后，患者可能会出现轻微的不适、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：每名患者大约采集15-20毫升血液 肿瘤样本：如果进行手术，切除的肿瘤的一段(约250-500毫克)将被快速冷冻并储存在-80摄氏度)以供进一步分析。 样品处理和数据分析：RNA样品和蛋白质样品将从FNA和组织样品中提取并保存在-80℃，直到分别使用芯片芯片SELDI-TOF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.