Magnetic Enrichment for Genetic Detection of Carcinoma Cells in Sputum

描述（由申请人提供）： 肺癌是美国的头号癌症杀手。发展敏感、无创的肺癌早期检测方法将降低肺癌的死亡率，是临床的重要目标。为了实现这一目标，我们已经确定了一组可以在痰液中检测到的遗传特征，为肺癌的早期检测提供了一个潜在的非侵入性生物标志物面板（Clin. Cancer Res，AACR Breaking News）。然而，遗传生物标志物在临床环境中的应用受到痰液细胞异质性的限制，痰液通常包括超过97%的巨噬细胞和中性粒细胞，而仅包括1%的支气管上皮细胞。新开发的磁性细胞分选（MACS）允许在单个过程中分离多达109个细胞，从而可以有效地从生物流体中富集稀有细胞用于分子遗传分析。R03研究的目的是通过检测痰液中富集的支气管上皮细胞中的遗传生物标志物来开发一种非侵入性诊断测试，用于早期检测肺癌，具有高灵敏度和特异性。我们建议：1）通过使用具有抗体珠的MACS从痰液中浓缩和纯化支气管上皮细胞以特异性地消耗巨噬细胞和嗜中性粒细胞，和2）确定生物标志物组用于早期检测来自I期肺癌患者的富集痰液样品中的肺癌的诊断性能。完成后，我们将开发一种遗传方法，用于有效检测痰中的生物标志物组，可用于高灵敏度的肺癌早期检测。这项R03研究将为未来的大规模队列和多中心研究奠定基础，这些研究旨在验证其在常规临床环境中的实用性。有助于实现我们目标的资源包括基础和临床研究人员的密切合作，以及分子遗传学，细胞病理学，肿瘤学和生物统计学方面的互补专业知识。此外，经批准的机构审查委员会方案将允许我们从早期肺癌患者和对照组中收集痰液样本，这将确保适当的分析并验证项目中开发测试的临床适用性。

DESCRIPTION (provided by applicant): Lung cancer is the number one cancer killer in the USA. The development of sensitive and noninvasive approaches for lung cancer early detection will reduce the mortality, is thus an important clinical goal. Toward this goal, we have identified a set of genetic signatures that can be detected in sputum, providing a potential noninvasive biomarker panel for the early detection of lung cancer (Clin. Cancer Res, AACR Breaking News). However, the application of the genetic biomarkers in the clinical settings is limited by the cellular heterogeneity of sputum, which typically includes more than 97% macrophages and neutrophils and only 1% bronchial epithelial cells. The newly developed magnetic cell sorting (MACS) allows up to 109 cells separated in a single process, thus can efficiently enrich rare cells from biological fluids for molecular genetic analysis. The objective of this R03 study is to develop a noninvasive diagnostic test by detecting the genetic biomarkers in enriched bronchial epithelial cells from sputum for the early detection of lung cancer with high sensitivity and specificity. We propose to: 1) concentrate and purify bronchial epithelial cells from sputum by using MACS with antibody-beads to specifically deplete macrophages and neutrophils, and 2) determine diagnostic performance of the biomarker panel for the early detection of lung cancer in the enriched sputum samples from patients with stage I lung cancer. At completion, we will develop a genetic approach for effectively detecting the biomarker panel in sputum that can be used for the early detection of lung cancer with high sensitivity. This R03 research will set the stage for future large-scale cohort and multicenter studies designed to validate its utility for adoption in routine clinical settings. Resources that will contribute towards our objective include close collaboration of basic and clinical investigators with complementary expertise in molecular genetics, cytopathology, oncology, and biostatistics. Furthermore, an approved Institutional Review Board protocol will allow us to collect sputum samples from early stage lung cancer patients and controls, which will secure appropriate analysis and verify clinical applicability of the developing test in the project.