High-throughput molecular-specific cell nanocytology for cancer screening

项目摘要 尽管人们普遍认为早期癌症筛查可以显著提高生存率，但目前还没有任何检测方法。 存在的主要癌症的准确和成本效益的筛查。一个值得注意的例外是宫颈癌， 由于巴氏涂片的引入，已从妇女癌症死亡的第一位降至第14位， 涂片作为初始筛查测试，然后是更具侵入性的阴道镜检查。然而，没有这样的两层 筛查模式可用于其他癌症。最近，我们在西北大学的研究小组， 与Northshore大学卫生系统合作，开发了一种新的癌症筛查模式， 癌性肿瘤的存在通过来自替代物的组织学正常细胞的简单拭子来检测 通过分波光谱（PWS）显微镜鉴定现场致癌作用。领域 致癌作用假定导致局灶性肿瘤的遗传/环境环境不仅存在于 这是一个特定的位置，但会影响整个器官。我们对750多名患者的数据显示， 对从这些替代部位的拭子中获得的细胞进行分析（例如，口腔细胞、肺癌、直肠癌 结肠癌等）可以区分非癌症和癌症患者。虽然这些数据 支持PWS对现场致癌作用敏感的观点，但它对那些 在癌症发展/进展中起关键作用的大分子/细胞器（例如染色质 复合物）。我们最近的数据表明，组织学染料可以提供PWS的分子特异性， 从特定细胞器或大分子结构散射的目标放大。因此，在这 我们将首先开发一种新的高通量分子PWS（HT-mPWS）仪器， 对细胞内细胞器具有分子特异性和纳米级敏感性。我们接下来将确定 细胞器/大分子在现场致癌过程中经历纳米级的最大变化。 由于将有许多染料可以针对特定的细胞器，我们将确定一个最佳的染料， 从靶细胞器散射的>10倍增强。虽然HT-mPWS是一种平台技术， 我们的首个测试项目将集中于肺癌筛查。肺癌 是美国人癌症死亡的主要原因，吸烟是最重要的风险因素 约占癌症的90%。我们对肺癌筛查的方法将是确定是否存在或 通过组织学正常的口腔细胞的简单拭子检查没有肺肿瘤。因此，我们将选择最佳 这种染料能最大限度地提高口腔细胞对肺癌的敏感性。我们接下来将测试HT-mPWS系统， 最佳染料和相应的分子标记物对100名患者的训练集和预测 随后将制定诊断肺癌患者的规则。最后，我们将测试预测规则， 一项对200名患者进行的独立盲法研究。这一提案的成功完成将导致一个多- 中心临床试验，随后临床实施HT-mPWS以筛查主要癌症。

Project Summary Although it is well accepted that early cancer screening would increase survival dramatically, no test currently exists for accurate and cost-effective screening of major cancers. A notable exception is cervical cancer, which has been relegated from the 1st to the 14th cause of cancer deaths in women due to the introduction of the Pap- smear as an initial screening test that is followed by a more invasive colposcopy. However, no such two-tier screening paradigm is available for other cancers. Recently our group at Northwestern University, in collaboration with Northshore University Health System, developed a new cancer screening paradigm in which the presence of a cancerous tumor is detected by a simple swab of histologically normal cells from surrogate sites via Partial Wave Spectroscopic (PWS) microscopy identification of field carcinogenesis. Field carcinogenesis posits that the genetic/environmental milieu that leads to a focal tumor exists not only at that particular location, but affects the entire organ. Our data from more than 750 patients showed that the PWS analysis of the cells obtained from a swab from these surrogate sites (e.g, buccal cells ¿ lung cancer, rectal cells ¿ colon cancer, etc.) could discriminate between non-cancerous and cancer patients. While these data support the idea that PWS is sensitive to field carcinogenesis, it lacks specificity to those macromolecules/organelles that play a critical role in cancer development/progression (e.g, chromatin complexes). Our recent data demonstrated that histological dyes can provide molecular specificity to PWS by the targeted amplification of scattering from specific organelles or macromolecular structures. Hence, in this proposal we will first develop a new high-throughput molecular PWS (HT-mPWS) instrument that will be both molecular-specific and nanoscale-sensitive to the intracellular organelle. We will next identify the organelles/macromolecules that undergo the greatest change in the nanoscale during field carcinogenesis. Since there will be many dyes that can target a specific organelle, we will identify an optimal dye that will give >10 fold enhancement of scattering from a targeted organelle. Although HT-mPWS is a platform technology to screen for different types of cancers, our initial test case will be focused on lung cancer screening. Lung cancer is the leading cause of cancer deaths among Americans, with smoking being the most important risk factor accounting for ~90% of cancers. Our approach to lung cancer screening will be to identify the presence or absence of a lung tumor by a simple swab of histologically normal buccal cells. Hence, we will select optimal dyes that will maximize the sensitivity of buccal cells to lung cancer. We will next test the HT-mPWS system, the optimal dyes and the corresponding molecular markers on a training set of 100 patients and a prediction rule to diagnose lung cancer patients will be subsequently developed. Finally, we will test the prediction rule on an independent blinded study of 200 patients. The successful completion of this proposal will lead to a multi- center clinical trial and subsequently to clinical implementation of HT-mPWS for screening major cancers.