High-throughput molecular-specific cell nanocytology for cancer screening

描述（由申请人提供）：尽管人们普遍认为早期癌症筛查可以显著提高生存率，但目前还没有一种检测方法可以准确、经济地筛查主要癌症。一个明显的例外是宫颈癌，由于采用了巴氏涂片检查作为最初的筛查试验，然后进行侵入性更强的阴道镜检查，宫颈癌已从妇女癌症死亡原因的第1位降至第14位。然而，对于其他癌症，没有这样的双层筛选范例。最近，我们在西北大学的研究小组与北岸大学卫生系统合作，开发了一种新的癌症筛查模式，通过局部波光谱（PWS）显微镜鉴定现场癌变，通过简单的拭子从替代部位提取组织学正常细胞来检测癌性肿瘤的存在。现场癌变假定导致局灶性肿瘤的遗传/环境环境不仅存在于该特定部位，而且影响整个器官。来自750多名患者的数据表明，从这些替代部位（例如，口腔细胞-肺癌，直肠细胞-结肠癌等）的拭子中获得的细胞的PWS分析可以区分非癌性和癌性患者。虽然这些数据支持PWS对现场癌变敏感的观点，但它对那些在癌症发生/进展中起关键作用的大分子/细胞器（例如染色质复合物）缺乏特异性。我们最近的数据表明，组织学染料可以通过特定细胞器或大分子结构的靶向扩增散射来提供PWS的分子特异性。因此，在本提案中，我们将首先开发一种新的高通量分子PWS （HT-mPWS）仪器，该仪器将对细胞内细胞器具有分子特异性和纳米级敏感性。接下来，我们将确定在现场致癌过程中，在纳米尺度上经历最大变化的细胞器/大分子。由于将有许多染料可以靶向特定的细胞器，我们将确定一种最佳染料，它将使>从目标细胞器散射增强10倍。虽然HT-mPWS是一种筛查不同类型癌症的平台技术，但我们最初的测试案例将集中在肺癌筛查上。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90%的癌症。我们的肺癌筛查方法是通过简单的拭子检测组织学上正常的颊细胞来确定是否存在肺肿瘤。因此，我们将选择最佳的染料，以最大限度地提高口腔细胞对肺癌的敏感性。接下来我们将对HT-mPWS系统进行测试，并在100例患者的训练集上测试最优的染料和相应的分子标记，随后将开发出诊断肺癌患者的预测规则。最后，我们将在200例患者的独立盲法研究中检验预测规则。该提案的成功完成将导致多中心临床试验，并随后将HT-mPWS用于筛查主要癌症的临床实施。

DESCRIPTION (provided by applicant): 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 i 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.