Differentiating Tumor Classes by Analysis of Alternative Splice Variant Patterns

描述(申请人提供)：随着基因组序列的完成和大约20-25,000个基因的完整组成，研究人员已经清楚地知道，人类的复杂性不能用这么少的基因来完全解释。MRNA的选择性剪接提供了一种复杂的方法，通过这种方法，单个基因可以以不同的亚型表达，正是通过这个过程，每个mRNA可以被翻译成几种不同的蛋白质。因此，这种复杂性为识别差异表达的基因异构体和潜在的诊断标记提供了丰富的来源。由于临床医生和外科医生不能在术前或术中确定恶性病变，所以细针抽吸(FNA)不确定的甲状腺病变的患者不能得到最佳的临床处理。因此，迫切需要更多的恶性肿瘤诊断标记物。因此，我们建议通过剪接阵列分析差异表达的选择性剪接变体来检查甲状腺肿瘤，通过RTPCR、原位杂交或免疫组织化学来验证我们的发现，并测试我们的方法在诊断不确定的甲状腺结节的FNA活检中的适用性。使用这种新的分子方法区分良性和恶性甲状腺肿瘤的能力可以改善美国每年超过10万名新患者的临床和外科治疗，这些患者的甲状腺FNA样本不确定或不充分。事实上，这种分析的潜在应用可以极大地改善这些患者的临床管理。公共卫生相关性：在美国，每年进行350,000次细针活检(FNA)以诊断甲状腺结节，其中超过100,000次是不确定的或不足以诊断的。因此，我们将研究这些肿瘤以区分分子标记。对于有这些FNA诊断的患者，使用区分分子模式可以极大地改善他们的外科治疗，因为在恶性肿瘤的情况下不需要两次手术，或者在良性肿瘤的情况下不需要患者进行任何手术。

DESCRIPTION (provided by applicant): With completion of the genome sequence and the estimated 20-25,000 genes comprising its entirety, it has become clear to investigators that the complexity of humans cannot be fully explained by so few genes. Alternative splicing of mRNA provides an intricate means by which individual genes can be expressed in various isoforms and it is through this process that each mRNA can be translated into several different proteins. This complexity thus provides a rich source for identifying differentially expressed gene isoforms and thus, potential diagnostic markers. Because the clinician and surgeon cannot determine malignancy pre- or intra-operatively, patients with indeterminate thyroid lesions on fine needle aspiration (FNA) cannot be optimally clinically managed. Therefore, additional diagnostic markers of malignancy are greatly needed. We therefore propose to examine thyroid tumors by splice array analysis for differentially expressed alternative splice variants, validate our findings by RTPCR, in situ hybridization, or immunohistochemistry and test the applicability of our assay on FNA biopsies form thyroid nodules that are indeterminate diagnostically. The ability to distinguish benign from malignant thyroid tumors using this novel molecular approach could improve both the clinical and surgical management of over 100,000 new patients in the United States annually who present with indeterminate or inadequate thyroid FNA samples. Indeed, potential application of this assay could dramatically improve the clinical management of these patients. PUBLIC HEALTH RELEVANCE: 350,000 fine needle aspirations (FNA) are performed annually in the United States to diagnose thyroid nodules, and of these, over 100,000 are indeterminate or inadequate for diagnosis. We therefore will study these tumors for differentiating molecular markers. For patients with these FNA diagnoses, use of differentiating molecular patterns could dramatically improve their surgical management by obviating the need for two operations in the case of a malignancy or, by obviating the need a patient undergoing any surgery in the case of a benign tumor.