Novel Surfaces for Enriching Epithelial Cells: A Breastmilk Model

描述（由申请人提供）：从体液中收获和浓缩上皮细胞是一项具有挑战性（昂贵且耗时）但重要的任务，这是癌症早期检测和治疗中的转化进展所需的，因为许多癌症类型涉及上皮细胞。例如，一种基于母乳中发现的乳腺上皮细胞DNA变化的非常有前途和准确的乳腺癌风险检测方法需要纯化和浓缩这些细胞，并排除牛奶中普遍存在的淋巴细胞，脂肪和蛋白质。由于必需的立即上皮富集过程在劳动力、仪器和实验室用品（抗体包被的珠粒）方面是昂贵的，因此需要上皮细胞收集和浓缩的替代方法。拟议的计划将开发能够选择性地粘附来自牛奶的上皮细胞并拒绝不需要的细胞组分和分子的表面。这种分离方案的潜在影响来自于表面本身的创新特征：采用坚固且经济的纳米级粘合剂元件，如聚合物线圈和固定化纳米颗粒，而不是昂贵且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体设备或珠子上，可以由缺乏高度专业培训的个人在现场使用。这些创新的表面使筛查方法成为可能，预计将具有广泛的可及性，因此对乳腺癌的早期检测产生广泛的积极影响。与此同时，在该计划中开发的表面设计策略可以通过微小的修改应用于创建针对其他上皮组织中的癌症和癌前细胞变化的额外表面，这些上皮组织从尿液，唾液和粘液中收集。 公共卫生相关性：该提案将开发新的表面，选择性地粘附悬浮液和人类母乳中的上皮细胞，以便随后分析乳腺癌或癌症风险的分子指标。这些表面是创新的，因为它们将使用固定的纳米级粘合元件而不是生物分子片段或抗体来实现靶细胞的选择性结合。该提案中开发的表面可以针对除乳汁外的其他体液进行改进，并为扩大便携式细胞富集技术奠定基础，从而增加更广泛女性人群的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.