Non-invasive Biomarker Panels for Early Detection of Lung Squamous Cell Carcinoma

描述（由申请人提供）：本申请是针对PA-09-199“鉴别癌症早期检测和预防的非编码RNA（ncRNA）靶点（R 01）"提交的。我们的目标是开发基于肺癌的小ncRNA生物标志物，可以与基因组探针联合收割机结合，以提高肺鳞状细胞癌（LSqCC）的早期检测。LSqCC是第二常见的肺癌。2010年，LSqCC将导致47，190人死亡，占美国所有癌症死亡人数的8.4%。鉴于晚期LSqCC预后不良，其早期发现将可能降低死亡率。然而，目前用于LSqCC早期检测的诊断技术要么是侵入性的，要么准确性差。我们收集了早期LSqCC患者和无癌对照的诱导痰样本。我们开发了一种富集技术，可以有效地收集深呼吸道上皮细胞从痰液样本。我们还开发了基于痰液的基因组探针，与痰液细胞学检查相比，可以更高的灵敏度诊断LSqCC。然而，探针的灵敏度不足以用于临床应用。小ncRNA正在成为癌症诊断中的潜在生物标志物。我们是第一个证明miRNAs稳定存在于痰液中的人。此外，使用微阵列分析42个早期LSqCC组织和配对的非癌肺组织，我们最近鉴定了39个LSqCC相关的小ncRNA。ncRNA包括30种miRNAs和9种snoRNAs，为恶性肿瘤的诊断提供了新的潜在生物标志物。因此，我们假设将ncRNA生物标志物与痰中的基因组探针相结合将改善LSqCC的早期检测。利用我们对病例和对照的诱导痰样本进行充分表征的优势，我们将执行以下四个具体目标：1）确定是否可以通过qRT-PCR在痰液中准确和稳健地测量miRNA和snoRNA，2）在痰液样本的训练集中优化用于早期LSqCC的一组高度特异性和灵敏性的ncRNA生物标志物，3），验证样品测试组中的ncRNA生物标志物，和4）评估ncRNA生物标志物和基因组探针用于LSqCC早期检测的组合使用。生物标志物在临床环境中的未来应用将可能提供一种有用的手段，以提高LSqCC的早期检测，从而降低死亡率。该研究将有力地支持PA的主要兴趣之一“体液中ncRNA和遗传变异标记的组合，以提高癌症诊断的准确性”。 公共卫生相关性：2010年，肺鳞状细胞癌（LSqCC）在美国估计导致47，190例死亡。我们将开发新的生物标志物，可用于非侵入性地识别早期LSqCC。生物标志物的未来应用将有可能降低LSqCC的死亡率。

DESCRIPTION (provided by applicant): This application is submitted in response to PA-09-199 "Identifying non-coding RNA (ncRNA) targets for cancer early detection and prevention (R01)". Our objective is to develop sputum-based small ncRNA biomarkers that can combine with the genomic probes to improve early detection of lung squamous cell cancer (LSqCC). LSqCC is the second most frequent lung cancer. In 2010, LSqCC will cause an estimated 47,190 deaths, accounting for 8.4 % of all cancer deaths in the USA. Given the poor prognosis associated with advanced stage LSqCC, its early detection will potentially reduce the mortality. However, the current diagnostic techniques for LSqCC early detection are either invasive or have poor accuracy. We have collected induced sputum samples from early stage LSqCC patients and cancer-free controls. We developed an enrichment technique that can efficiently collect deep respiratory epithelial cells from the sputum samples. We also developed sputum-based genomic probes that can diagnose LSqCC with higher sensitivity compared with sputum cytology. However, the sensitivity of the probes is not high enough for clinical application. Small ncRNAs are emerging as potential biomarkers in cancer diagnosis. We have been the first to demonstrate that miRNAs are stably present in sputum. Furthermore, using microarrays to analyze 42 early stage LSqCC tissues and paired noncancerous lung tissues, we recently identified 39 LSqCC-associated small ncRNAs. The ncRNAs include 30 miRNAs and 9 snoRNAs, providing new potential biomarkers for diagnosis of the malignancy. Therefore, we hypothesize that combining the ncRNA biomarkers with the genomic probes in sputum will improve early detection of LSqCC. Taking advantage of our well-characterized induced sputum samples of the cases and controls, we will perform the following four specific aims: 1), determining whether the miRNAs and snoRNAs can be accurately and robustly measured in sputum by qRT-PCR, 2), optimizing a panel of highly specific and sensitive ncRNA biomarkers for early stage LSqCC in a training set of the sputum specimens, 3), validating the ncRNA biomarkers in an testing set of the samples, and 4), evaluating the combined use of the ncRNA biomarkers and genomic probes for early detection of LSqCC. Future application of the biomarkers in clinical settings will potentially provide a useful means to improve early detection of LSqCC, and hence reduce the mortality. The study will strongly support one of the PA's main interests "Combination of ncRNA and genetic variation markers in body fluids to increase accuracy of cancer diagnosis". PUBLIC HEALTH RELEVANCE: Lung squamous cell cancer (LSqCC) causes an estimated 47,190 deaths in 2010 in the USA. We will develop new biomarkers that can be used to noninvasively identify early stage LSqCC. Future application of the biomarkers will have the potential to reduce the mortality of LSqCC.