New Diagnostic Assay for Early Detection of Lung Cancer

描述（申请人提供）：早期诊断肺癌可降低与肺癌相关的死亡率。我们已经证明，染色体探针的多色荧光原位杂交在检测肺癌细胞时比传统的细胞学研究更敏感。我们精确地定义了原发性肺癌的基因组特征，并为这些特征开发了特异性探针。我们还根据机构审查委员会批准的方案获得了肺癌患者的生物样本和相应的流行病学和临床数据。我们假设这些特征构成了生物标志物，并且一组针对基因组特征的特异性探针的原位策略将足够敏感和特异性地检测早期肺肿瘤。基于原位杂交的原理，利用阵列技术的优势，我们建议开发一种创新的原位阵列，它将允许同时测量这些关键的肿瘤特异性遗传变化，并且可以简单地应用于易于获取的替代标本。我们将首先通过肺组织微阵列的双色荧光原位杂交来评估潜在的生物标志物，以确定一组为检测肺癌细胞提供最大总体灵敏度和特异性的探针。然后，我们将开发一种原位基因组DNA阵列：一个玻璃盖盖，上面将结合用多色荧光染料标记的这些探针的混合物。我们将用连续接受开胸手术的肺癌患者的痰液、口腔刷牙和肺肿瘤组织标本来测试该方法，并将这些数据与临床协变量进行比较，以确定该方法对替代组织的诊断能力。最后，我们将结合自动点计数系统，开发一种可靠、经济、快速的肺癌基因检测方法。这项研究将导致第一个基于原位的检测，由多个特定的基因探针组成，可用于替代材料的常规诊断。这一新策略将对大规模基于人群的分子流行病学研究和治疗干预具有重大意义。当与其他适当的肿瘤特异性基因结合时，它也将在其他类型癌症的诊断和分期方面有很大的应用。这个项目将是多学科的，包括流行病学、分子生物学、细胞遗传学、病理学、生物信息学和统计学。

DESCRIPTION (provided by applicant): Early diagnosis of lung cancer may reduce the mortality associated with it. We have demonstrated that multicolor-fluorescence in situ hybridization with chromosomal probes is more sensitive in detecting lung cancer cells than conventional cytologic studies are. We precisely defined genomic signatures in primary lung cancer and developed specific probes for these signatures. We also obtained biologic samples and corresponding epidemiologic and clinical data from patients with lung cancer under an Institutional Review Board approved protocol. We hypothesize that these signatures constitute biomarkers and that an in situ strategy with a panel of specific probes for the genomic signatures will be sufficiently sensitive and specific to detect early neoplasia of the lung. Based on the principle of in situ hybridization, taking advantage of array techniques, we propose to develop an innovative in situ array that will allow simultaneous measurement of these key tumor-specific genetic changes and that can be simply applied to easily accessible surrogate specimens. We will first evaluate the potential biomarkers by dual-color fluorescence in situ hybridization with lung tissue microarrays to identify a set of probes that provide the greatest overall sensitivity and specificity for detection of lung cancer cells. We will then develop an in situ genomic DNA array: a glass coverslip to which a cocktail of these probes, labeled with multicolor fluorochromes, will be bound. We will test the assay with sputum and oral brushings and lung tumor tissue specimens from consecutive patients with lung cancer who had undergone thoracotomy and compare these data with clinical covariates to determine the assay's diagnostic ability with surrogate tissues. Finally, we will combine the assay with an automated dot-counting system to develop a reliable, cost-effective, rapid genetic test for lung carcinogenesis. This study should result in the first in situ-based assay consisting of multiple specific genetic probes that can be employed for routine diagnosis with surrogate materials. This novel strategy will have substantial implications for large-scale population-based molecular epidemiologic studies and therapeutic interventions. It will also have great application for the diagnosis and staging of other types of cancer when combined with other appropriate tumor-specific genes. This project will be multidisciplinary, incorporating epidemiology, molecular biology, cytogenetics, pathology, bioinformatics, and statistics.