Nanoparticle-based Mannetic Microluidic Enrichment System (MMES)

描述（由申请人提供）：富集的异常细胞将使基因组、蛋白质组和表观基因组分析能够检测表达基因、蛋白质/肽谱和表观遗传标记物中的癌症特异性改变。使用体液以常规方式高效地进行这种富集的能力可能导致早期癌症检测的改进。目前的富集方法依赖于荧光激活细胞分选（FACS）和免疫磁性分离（IMS）。流式细胞术是一个新兴的诊断领域，它提供了高纯度的细胞亚群的分离，然而，它的分选速度（104个细胞/秒）是缓慢的，它的设备是大的和昂贵的。IMS是一种简单、低成本的分离靶细胞的方法，但批量操作限制了其通量，且富集因子较低。该NIH SBIR I期提案旨在开发一种简单、高通量和高富集因子的系统，用于富集罕见癌细胞。在第一阶段，我们将证明使用尺寸可调的氧化铁磁性纳米颗粒（MNP）富集具有高富集因子的癌细胞的可行性。在第二阶段，我们将通过关注体液中的特定癌细胞来优化该系统。本项目的最终成果将包括无柱试剂盒、柱试剂盒和用于不同富集水平的磁性微流控浓缩器。 公共卫生相关性：超过80%的人类肿瘤起源于上皮细胞，通常在粘膜表面，并且是克隆起源（例如，结肠、肺、前列腺、口腔、食道、胃、子宫颈、膀胱）。当肿瘤细胞自发脱落到痰、尿甚至各种渗出液中时，它们的发展往往变得明显。这些脱落细胞内的分子和遗传异常可用于检测和识别癌前病变或非常早期的癌症，如果临床上可获得高灵敏度的技术来识别数百万正常细胞中的少数异常细胞。该项目的成功开发将为富集体液中的罕见癌细胞和细胞碎片提供高通量和高富集因子系统，用于早期癌症检测。

DESCRIPTION (provided by applicant): Enriched abnormal cells will enable genomic, proteomic, and epigenomic analyses to detect cancer-specific alterations in expressed genes, protein/peptide profiles, and epigenetic markers. The ability to carry out such enrichment in high efficiency in a routine way using body fluids may lead to improvements in cancer detection at early stage. The current enrichment methods rely on fluorescence-activated cell sorting (FACS) and immunomagnetic separation (IMS). FACS is an emerging diagnostic field which offers the isolation of cell subsets with high purity however, its sorting rate (104 cells/s) is slow and its apparatus is large and expensive. IMS is simple and low cost method to separate targeting cells however, the batch operation limits its throughput and its enrichment factor is low. This NIH SBIR Phase I proposal is to develop a simple, high throughput and high enrichment factor system for the enrichment of rare cancer cells. In phase I, we will demonstrate the feasibility of using size tunable iron oxide magnetic nanoparticles (MNPs) for the enrichment of cancer cells with high enrichment factor. In phase II, we will optimize this system by focusing on a specific cancer cell in body fluid. The final deliverables of this project will include columnless kit, column kit, and the magnetic microfluidic concentrator for different level of enrichment. PUBLIC HEALTH RELEVANCE: More than 80 percent of human tumors originate from epithelial cells, often at a mucosal surface, and are clonal in origin (e.g., colon, lung, prostate, oral cavity, esophagus, stomach, uterine cervix, bladder). Their development often becomes apparent when tumor cells exfoliate spontaneously into sputum, urine, or even into various effusions. The molecular and genetic abnormalities within these exfoliated cells could be used to detect and identify precancerous lesions or very early stage cancer if highly sensitive technologies were clinically available to identify the few abnormal cells among millions of normal cells. Successful development of this project will provide a high throughput and high enrichment factor system for the enrichment of rare cancer cells and cell fragments in body fluids for early cancer detection.