Novel Surfaces for Enriching Epithelial Cells: A Breastmilk Model

描述(申请人提供)：从体液中采集和浓缩上皮细胞是一项具有挑战性的(昂贵和耗时的)任务，但由于许多癌症类型涉及上皮细胞，因此需要在癌症的早期发现和治疗方面取得翻译进展。例如，一种非常有希望和准确的乳腺癌风险检测方法，基于在母乳中发现的乳房上皮细胞DNA的变化，需要对这些细胞进行纯化和浓缩，并排除牛奶中普遍存在的淋巴细胞、脂肪和蛋白质。由于必要的即刻上皮细胞浓缩过程在人工、仪器和实验室用品(抗体包被的珠子)方面代价高昂，因此需要替代的上皮细胞收集和浓缩方法。拟议中的计划将开发能够选择性地附着牛奶中的上皮细胞并拒绝不需要的细胞部分和分子的表面。这种分离方案的潜在影响来自于表面本身的创新特征：使用了坚固而经济的纳米级粘连元件，如聚合物线圈和固定的纳米颗粒，而不是昂贵而脆弱的生物分子片段和抗体。这些复杂的表面可以部署在高度敏感的微流控设备中或珠子上，这些微流控设备可以由缺乏高度专业培训的个人在现场使用。由这些创新表面实现的筛查方法预计将具有广泛的可及性，从而对乳腺癌的早期检测产生广泛的积极影响。同时，在这个项目中开发的表面设计策略，只要稍加修改，就可以应用于创建针对从尿液、唾液和粘液中收集的其他上皮组织中的癌症和癌前细胞变化的额外表面。 公共卫生相关性：这项提议将开发新的表面，从悬浮液和人类母乳中选择性地附着上皮细胞，以使它们能够随后对乳腺癌或癌症风险的分子指标进行分析。这种表面是创新的，因为它们将使用固定的纳米级黏附元件而不是生物分子片段或抗体来实现对目标细胞的选择性结合。这项提议中开发的表面可以精炼成牛奶以外的其他体液，并为扩大便携式细胞浓缩技术奠定基础，这些技术增加了更广泛的女性人口对早期风险评估的可及性。

DESCRIPTION (provided by applicant): Harvesting and concentrating epithelial cells from body fluids is a challenging (expensive and time-consuming) but important task required for translational advances in the early-detection and treatment of cancer, since many cancer types involve epithelial cells. As an example, a highly promising and accurate risk detection method for breast cancer, based on changes in the DNA of breast epithelial cells found in human milk, requires purification and concentration of these cells, and exclusion of lymphocytes, fats, and proteins that are prevalent in milk. Because the necessary immediate epithelial enrichment process is costly in terms of labor, instrumentation, and lab supplies (antibody-coated beads), alternate methods of epithelial cell collection and concentration are needed. The proposed program will develop surfaces capable of selectively adhering epithelial cells from milk and rejecting the undesired cell fractions and molecules. The potential impact of this separation scheme comes from the innovative features of the surfaces themselves: robust and economical nano-scale adhesive elements such as polymer coils and immobilized nanoparticles are employed instead of expensive and fragile biomolecular fragments and antibodies. These sophisticated surfaces could be deployed in highly sensitive microfluidic devices or on beads, which could be used in the field by individuals who lack highly specialized training. The screening methods made possible by these innovative surfaces are expected to have wide accessibility and therefore broad positive impact for early detection of breast cancer. At the same time the surface design strategies developed in this program could, with minor modification, be applied to the creation of additional surfaces targeting cancer and pre-cancerous cell changes in other epithelial tissues, collected from urine, saliva, and mucus. PUBLIC HEALTH RELEVANCE: This proposal will develop novel surfaces that selectively adhere epithelial cells from suspensions and human breast milk, to enable their subsequent analysis for molecular indicators of breast cancer or cancer risk. The surfaces are innovative because they will achieve selective binding of targeted cells using immobilized nano-scale adhesive elements rather than biomolecular fragments or antibodies. The surfaces developed in this proposal can be refined for other body fluids besides milk, and form the foundation for scaled-up portable cell enrichment technologies that increase the accessibility of early risk assessment to a broader population of women.