Molecular Classification of Suspicious Thyroid Tumors

A.具体目标 可疑甲状腺结节的鉴别诊断难题。 虽然细针吸取细胞学(FNA)是鉴别诊断甲状腺结节的最佳诊断工具，但它通常不能区分良恶性病变。因此，大多数甲状腺结节患者因为可疑但不确定的恶性肿瘤诊断而需要手术。由于良性和恶性甲状腺结节的外科治疗不同，可疑FNAs的患者可能会以不太理想的手术方式进行治疗。一些良性病变的患者可能接受不必要的手术，而一些恶性病变的患者一旦在永久组织切片上被诊断为恶性肿瘤，可能需要再次手术才能完成甲状腺切除术。 研究前提 虽然其他人和我们已经记录了甲状腺肿瘤的各种基因改变，但没有特定的改变(S)可以可靠地区分良恶性病变。我们认为，特定的基因表达模式与不同的甲状腺肿瘤类型有关，这些独特的表达模式可以用于区分一种肿瘤类型和另一种肿瘤类型。因此，我们建议通过基因芯片分析良性和恶性甲状腺病变的基因分型，这些病变在FNA细胞学上可以而且经常表现为“可疑”病变。为此，我们对63个甲状腺肿瘤进行了微阵列分析，并在8个甲状腺肿瘤亚型中的4个中选择了一组最能区分良恶性的11个基因。这些数据将得到验证和扩展，以包括另外4种甲状腺肿瘤亚型。然后，一组候选的、可区分的基因将通过实时RT-PCR和甲状腺肿瘤的原位杂交(ISH)来验证，随后是组织芯片的免疫组织化学(IHC)和细胞块的免疫细胞化学(ICC)。 从甲状腺细胞系和FNA样本制备。]最终和未来的目标将是将这些结果应用于可疑甲状腺FNA样本的鉴别诊断，以改善对患有这种诊断的患者的临床管理。 为了实现这些目标，我们提出了以下建议： 具体目标1：通过基因芯片分析确定甲状腺肿瘤的良、恶性。 特异性目的2：用实时定量RT-PCR和原位杂交技术验证目的1中筛选的候选基因集。 具体目的3：建立免疫组织化学(IHC)分析AIM 2中确认的基因，通过组织微阵列分析(TMA)对甲状腺肿瘤和[从石蜡包埋的甲状腺细胞系制备的细胞块，并对从FNA样本制备的细胞块进行免疫细胞化学(ICC)分析]进行验证。

A. SPECIFIC AIMS Diagnostic Dilemma in the Differential Diagnosis of a Suspicious Thyroid Nodule. Although fine needle aspiration (FNA) cytology is the best diagnostic tool in the differential diagnosis of a thyroid nodule, it often cannot differentiate a benign from a malignant lesion. Consequently, a majority of patients with thyroid nodules require surgery because of a suspicious, but not definite, diagnosis of malignancy. Because the surgical management for benign and malignant thyroid nodules differs, patients with suspicious FNAs may be treated in a less than ideal way surgically. Some patients with benign lesions may undergo unnecessary surgery and some with malignant lesions may require a second operation for completion thyroidectomy once a diagnosis of malignancy is rendered on permanent histological section. Research Premise Although others and we have documented various genetic alterations in thyroid neoplasms, no specific alteration(s) can reliably distinguish benign from malignant lesions. We propose that specific gene expression patterns are associated with the various thyroid tumor types and that these unique expression patterns can be capitalized upon to differentiate one tumor type from another. We therefore propose to genotype by microarray analysis the benign and malignant thyroid lesions that can and often do present as "suspicious" lesions on FNA cytology. [To that end we have performed microarray analysis on 63 thyroid tumors and have selected a set of 11 genes that best discriminate benign from malignant in 4 of 8 thyroid tumor sub-types. These data will be validated and expanded to include 4 additional thyroid tumor sub-types. A panel of candidate, discriminating genes will then be validated by real time RT-PCR and in situ hybridization (ISH) of thyroid tumors, followed by both immunohistochemistry (IHC) of tissue microarrays and, immunocytochemistry (ICC) of cell blocks Prepared from thyroid cell lines and FNA samples.] The ultimate and future goal would then be to apply these results to the differential diagnosis of suspicious thyroid FNA samples in order to improve the clinical management of patients with this diagnosis. To accomplish these goals we propose the following: Specific Aim 1: To genomically define by microarray analysis benign and malignant thyroid tumors. Specific Aim 2: To validate the candidate gene sets selected in Aim 1 by real-time quantitative RT-PCR and, [in situ hybridization ISH]. Specific Aim 3: To develop immunohistochemical (IHC) assays for genes validated in Aim 2, validate these by tissue microarrays analysis (TMA) of thyroid tumors and [cell blocks prepared from paraffin-embedded thyroid cell lines, and perform immunocytochemical (ICC) assays to cell blocks prepared from FNA samples.]