CAREER: Label-Free Cell Sorting Based on Single-Cell Metabolism

该项目将开发选择性分离癌细胞的技术。这将允许在单细胞水平上进行表征，从而实现有效的靶向癌症治疗。结合相关技术的实验室模块将被设计为提供鼓舞人心的、基于发现的体验，这些体验已被证明对指导和留住科学和工程领域的学生至关重要。这项工作将在一个主要是本科生的机构进行，在那里本科生接触到真实的研究经验将吸引不同的学生。实验包和教材将通过已建立的项目和网络每年提供给当地社会经济条件较弱的初中和高中的数百名学生。通过原始材料、网站和视频广泛传播这些模块将鼓励培养有能力的生物技术工作人员。高糖代谢是癌症的一个标志。癌细胞会将葡萄糖转化为乳酸。肿瘤中高水平的乳酸与转移、复发和不良预后相关。因此，表现出高乳酸分泌率的细胞是治疗的特定目标。对这些重要而稀有的细胞的研究需要有效的分离工具。一种新型的无标记微流控平台将用于基于细胞代谢的单细胞快速和廉价的分离。单细胞全转录组分析将以定量的方式将基因表达与细胞表型联系起来。它将揭示细胞间基因表达的异质性，并阐明高度恶性癌细胞的决定因素。由于糖酵解是细胞活力的明确指标，该平台还将用于快速简便地分离活细胞和死细胞，用于药理学和毒理学应用。最终，本提案中开发的技术可以应用于基于细胞代谢的细胞或亚群分离，在细胞生物学，生物能源，生物技术和医学中有广泛的应用。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

The project will develop technology to selectively isolate cancer cells. This will allow characterization at the single cell level to enable effective, targeted cancer therapies. Laboratory modules that incorporate relevant technologies will be designed to provide inspirational, discovery-based experiences that have been shown to be critical for both directing and retaining students in science and engineering fields. This work will be performed at a primarily undergraduate institution, where exposure of undergraduates to authentic research experiences will engage a diverse cross-section of students. Experimental kits and teaching material will be developed and offered through established programs and networks to hundreds of students per year at local socioeconomically disadvantaged middle school and high schools. Wide dissemination of these modules via source material, website, and videos will encourage the development of a capable biotechnology workforce.High glucose metabolism is one hallmark of cancer. Cancer cells are programmed to convert glucose to lactate. High levels of lactate in tumors are associated with metastasis, recurrence, and poor outcomes. Cells that exhibit high lactate secretion rates are therefore specific targets for therapy. The study of these important and rare cells requires efficient tools for their isolation. A novel label-free microfluidic platform will be used for the fast and inexpensive isolation of single cells based on cellular metabolism. Single-cell whole transcriptome analysis will connect gene expression to cell phenotype in a quantitative fashion. It will reveal the cell-to-cell heterogeneity in gene expression and clarify the determinants of highly malignant cancer cells. As glycolysis is a clear indicator of cell vitality, the platform will also be used to quickly and easily separate live and dead cells for applications in pharmacology and toxicology. Ultimately, the technology developed in this proposal can be applied to isolate cells or subpopulations based on cellular metabolism for a broad range of applications in cell biology, bioenergy, biotechnology and medicine.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.