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

职业：基于单细胞代谢的无标记细胞分选

• 批准号: 1751861
• 负责人: Paul Abbyad
• 金额: $ 50.5万
• 依托单位: Santa Clara University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1751861&HistoricalAwards=false
• 关键词: CAREER; Label; Free; Cell; Sorting

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.

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、有针对性的癌症治疗。将设计包含相关技术的实验室模块，以提供鼓舞人心的、基于发现的经验，这些经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。这项工作将在一个以本科生为主的机构进行，在那里，本科生接触到真实的研究经验将吸引不同阶层的学生。将开发实验工具包和教材，并通过现有的计划和网络向当地社会经济困难的初中和高中每年数百名学生提供。通过原始材料、网站和视频广泛传播这些模块将鼓励有能力的生物技术工作人员的发展。高糖代谢是癌症的一个标志。癌细胞被编程将葡萄糖转化为乳酸。肿瘤中高水平的乳酸与转移、复发和不良预后有关。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工具来分离它们。一种新型的无标记微流控平台将用于基于细胞代谢的快速、廉价的单细胞分离。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，该平台还将被用于快速轻松地分离活细胞和死亡细胞，用于药理学和毒理学的应用。最终，这项建议中开发的技术可以应用于分离基于细胞代谢的细胞或亚群，用于细胞生物学、生物能源、生物技术和医学的广泛应用。这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

项目成果

Sorting by interfacial tension (SIFT): Label-free enzyme sorting using droplet microfluidics

• DOI: 10.1016/j.aca.2019.08.025
• 发表时间: 2019-12-16
• 期刊: ANALYTICA CHIMICA ACTA
• 影响因子: 6.2
• 作者: Horvath, Daniel G.;Braza, Samuel;Abbyad, Paul
• 通讯作者: Abbyad, Paul

Multiplexed Single-Cell Measurements of FDG Uptake and Lactate Release Using Droplet Microfluidics

• DOI: 10.1177/1533033819841066
• 发表时间: 2019-03-29
• 期刊: TECHNOLOGY IN CANCER RESEARCH & TREATMENT
• 影响因子: 2.8
• 作者: Sengupta, Debanti;Mongersun, Amy;Pratx, Guillem
• 通讯作者: Pratx, Guillem

Sorting by interfacial tension (SIFT): label-free selection of live cells based on single-cell metabolism

• DOI: 10.1039/c8lc01328d
• 发表时间: 2019-04-21
• 期刊: LAB ON A CHIP
• 影响因子: 6.1
• 作者: Pan, Ching W.;Horvath, Daniel G.;Abbyad, Paul
• 通讯作者: Abbyad, Paul