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

总结/摘要 肺癌是美国男性和女性癌症相关死亡率的主要原因， 5年生存率只有15%。虽然手术是唯一的治疗选择，但70%的患者被诊断为 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，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 通过LDCT识别的肺部病变最终被发现是良性的。因此，这些患者接受医学治疗。 不必要的程序和治疗，导致成本、时间和发病率增加。因此， 迫切需要开发一种非侵入性的补充筛查技术， 因此，能够在早期和潜在可治愈的阶段检测肺部病变。到 为此，在吸烟者痰液脱落的支气管上皮细胞中发现细胞学和分子畸变 与肺癌发病率有很强的相关性。然而，这些重要的诊断细胞 在痰液的细胞组成中占不到1%，因此其检测极具挑战性。 因此，开发一种能够分离和捕获这些稀少但重要的细胞的方法， 从痰中去除其他模糊细胞，从而提高这些重要细胞的临床效用， 生物标本第一阶段申请的目标是开发一种新型的肽涂层载玻片， 选择性地捕获和保留肽修饰区域中的感兴趣的上皮细胞，同时移动肽修饰区域中的上皮细胞。 将细胞遮蔽到载玻片的单独区域。肽修饰载玻片上的上皮细胞捕获将是第一次 优化以鉴定用于后续临床测试的先导肽。此后，肽修饰的化合物的相容性 将建立具有常用临床染色技术的载玻片。最后，分析了使用的可行性 这些用于从人痰标本中捕获细胞的肽修饰的载玻片将被验证。成功 该项目的完成将不仅允许临床医生更好地观察捕获的上皮细胞， 而且能够进一步下游处理这些细胞以鉴定存在的分子和/或遗传缺陷， 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.