Magnetic Enrichment for Genetic Detection of Carcinoma Cells in Sputum

描述(由申请人提供)： 肺癌是美国头号癌症杀手。发展敏感的、非侵入性的肺癌早期检测方法将降低死亡率，因此是一个重要的临床目标。为了这个目标，我们已经确定了一组可以在痰中检测到的基因特征，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小组(Clin)。癌症资源，AACR突发新闻)。然而，由于痰中含有97%以上的巨噬细胞和中性粒细胞，而支气管上皮细胞仅占1%，限制了遗传生物标志物在临床中的应用。最新开发的磁性细胞分选(MACS)允许在一个过程中分离多达109个细胞，从而可以有效地从生物液中浓缩稀有细胞，用于分子遗传分析。本R03研究的目的是通过检测痰中富集的支气管上皮细胞中的遗传生物标记物，建立一种无创诊断方法，用于肺癌的早期诊断，具有较高的敏感性和特异性。我们建议：1)通过使用带有抗体微球的MSC特异性地去除巨噬细胞和中性粒细胞来浓缩和纯化痰中的支气管上皮细胞；2)从I期肺癌患者的浓缩痰样本中确定用于早期检测肺癌的生物标志物组合的诊断性能。完成后，我们将开发一种基因方法，有效地检测痰中的生物标记物小组，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，旨在验证其在常规临床环境中采用的有效性。有助于实现我们目标的资源包括在分子遗传学、细胞病理学、肿瘤学和生物统计学方面具有互补专业知识的基础和临床研究人员的密切合作。此外，机构审查委员会批准的方案将允许我们收集早期肺癌患者和对照组的痰样本，这将确保进行适当的分析，并验证正在开发的测试在项目中的临床适用性。

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.