Peptide-based slides for improving the diagnostic quality of sputum specimens

摘要/摘要 肺癌是美国男性和女性癌症相关死亡的主要原因， 令人沮丧的5年存活率为15%。虽然手术是唯一的治愈选择，但70%的患者被诊断出 无法手术的晚期疾病，从而突出了缺乏有效的早期筛查方法。通常情况下，低 剂量计算机断层扫描(LDCT)被用作主要的筛查方式，然而，90%-95%的可疑 LDCT发现的肺部病变最终被发现是良性的。因此，这些患者在医学上接受了 不必要的程序和治疗，导致成本、时间和发病率增加。因此，有一种 迫切需要开发一种能够识别高危人群的无创补充筛查技术 这一技术有助于肺癌患者的早期诊断，从而能够在早期和潜在的可治愈阶段发现肺部病变。至 为此，在吸烟者的痰中发现了脱落的支气管上皮细胞的细胞学和分子异常。 已经显示出与肺癌发病率有很强的相关性。然而，这些具有诊断意义的细胞 只占痰中细胞成分的不到1%，因此对它们的检测极具挑战性。 因此，开发一种方法来分离和捕获这些稀少但重要的细胞是至关重要的，同时 从痰中去除其他遮蔽细胞，从而提高这些至关重要的 生物群落。这项第一阶段应用的目标是开发一种新型的多肽涂层玻璃片，它可以 选择性地捕获和保留多肽修饰区域中的感兴趣的上皮细胞，同时移动 将单元格遮挡到幻灯片的单独区域。在多肽修饰的玻片上捕获上皮细胞将是第一次 经过优化，可识别用于后续临床测试的铅多肽。此后修饰的多肽的配伍性 将建立具有临床常用染色技术的载玻片。最后，分析了使用的可行性。 这些用于从人类痰标本中捕获细胞的多肽修饰玻片将得到验证。成功 该项目的完成不仅将允许临床医生更好地可视化捕获的上皮细胞， 而且还允许对这些细胞的进一步下游处理以识别存在的分子和/或遗传异型性， 因为这项技术的非破坏性。

SUMMARY/ABSTRACT Lung cancer is the leading cause of cancer-related mortality in both men and women in United States with a dismal 5-year survival rate of 15%. While surgery is the only curative option, 70% of the patients are diagnosed with inoperable advanced disease, thus highlighting a lack of effective early screening methods. Typically, low dose computed tomography (LDCT) is used as the primary screening modality, however, 90-95% of suspicious lung lesions identified by LDCT eventually are found to be benign. As a result, such patients undergo medically unnecessary procedures and treatments, resulting in increased cost, time and morbidity. Hence there is an urgent need to develop a non-invasive complementary screening technique which can identify the high-risk population for lung cancer and thus enable detection of lung lesions at an early and potentially curable stage. To this end, cytological and molecular aberrations found in exfoliated bronchial epithelial cells in sputum of smokers have shown a strong correlation to lung cancer incidence. However, these diagnostically important cells represent less than 1% of the cellular composition of sputum, thus making their detection extremely challenging. Hence, it is essential to develop a method which can isolate and capture these scant yet important cells, while removing the other obscuring cells from sputum and thereby improve the clinical utility of these vital biospecimens. The goal of this Phase I application is to develop a novel peptide-coated glass slide, which can selectively capture and retain the epithelial cells of interest in a peptide-modified region, while moving the obscuring cells to a separate region of the slide. Epithelial cell capture on the peptide-modified slides will be first optimized to identify lead peptides for subsequent clinical testing. Thereafter compatibility of the peptide-modified slides with commonly employed clinical staining techniques will be established. Finally, the feasibility of using these peptide-modified slides for capturing cells from human sputum specimens will be validated. Successful completion of this project will not only allow for better visualization of the captured epithelial cells to the clinicians, but also enable further downstream processing of these cells to identify molecular and/or genetic atypia present, because of the non-destructive nature of this technology.