Understanding of therapeutic cells functions: A single-cell lab on a chip

了解治疗细胞功能：芯片上的单细胞实验室

• 批准号: 1803872
• 负责人: Tania Konry
• 金额: $ 29.95万
• 依托单位: Northeastern University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1803872&HistoricalAwards=false
• 关键词: Understanding; therapeutic; cells; functions; single

The human body is capable of killing cancer cells without the assistance of drugs. This occurs frequently and is credited to cells that are part of the immune system. They are referred to as natural killer (NK) cells. The mechanism of their interaction with cancer cells is not understood well. One major difficulty is that the interaction is occurring within the body, and we usually do not detect it until a large population of cancer cells (a tumor) is present. At that point, a large number of tumor cells is interacting with a large number of NK cells. A lot more could be learned about the functions of the NK cells if they were observed interacting individually with a single cancer cell. This project will design a device that should enable those observations to be made by creating droplets containing one NK cell and one cancer cell. The device will be used to measure several aspects of the interaction. It will also allow the observation of NK cells with cancer cells that are in many different stages of development, and activity in the presence of different drugs. If this project is successful, it could lead to dramatic improvements in strategies for cancer immunotherapy, leading to longer and healthier lives after a cancer diagnosis. In addition, this project will support research experiences for high school and college students. It will also train community college faculty in many of the research techniques used, greatly amplifying the positive impact on developing a highly skilled biotechnology workforce.The project will focus on a novel microfluidic chip that will integrate large arrays to guarantee sufficient number of reactions for screening, and parallel designs to achieve multiplexing capabilities. The parallel device designs will enable screening of different combinations of drugs/concentrations/cells across multiple storage arrays of bio-reactions. These designs simplify the number of inlets required for operation of the micro-chip and create micro-channels with unified flow resistance for synchronous generation of bioreactors with the same dimensions/volumes. Furthermore, the design incorporates an on-board gradient generator that creates a series of drug concentrations and enables testing of different therapeutic cell lines against one target specimen. The single cell resolution and the dynamic measurement of multiplex biomarkers allows the dynamic assessment of the specific state of any cell type before during and after therapy. This allows regulation mechanisms of activation and inhibition and the causality of events to be determined. Thus the proposed on-chip approach allows concurrent assessment of vaccine therapeutic potential, correlating mechanistic and functional information to gain greater insight into cell functions and responses at multiple levels. The methods developed by the proposed studies will further allow us to answer important biological questions. These include the contribution of stochasticity in regulating cell-based therapies, and cell responses and variation in response to a broad range of immunomodulatory stimuli.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.

人体能够在没有药物帮助的情况下杀死癌细胞。这种情况经常发生，并归因于免疫系统的一部分细胞。它们被称为自然杀伤细胞（NK）。它们与癌细胞相互作用的机制尚不清楚。一个主要的困难是这种相互作用发生在体内，我们通常在大量癌细胞（肿瘤）出现之前不会发现它。此时，大量肿瘤细胞与大量NK细胞发生相互作用。如果观察NK细胞单独与单个癌细胞相互作用，我们可以了解更多NK细胞的功能。该项目将设计一种设备，通过制造含有一个NK细胞和一个癌细胞的液滴来进行这些观察。该设备将用于测量交互的几个方面。它还将允许观察NK细胞与处于许多不同发展阶段的癌细胞，以及在不同药物存在下的活性。如果这个项目成功，它可能会导致癌症免疫治疗策略的巨大改进，导致癌症诊断后更长寿、更健康的生活。此外，该项目将为高中生和大学生提供研究经验。它还将培训社区学院的教师使用许多研究技术，大大扩大对发展高技能生物技术劳动力的积极影响。该项目将专注于一种新型微流控芯片，该芯片将集成大型阵列，以保证足够数量的反应进行筛选，并并行设计以实现多路复用能力。平行装置设计将能够筛选跨多个生物反应存储阵列的不同药物/浓度/细胞组合。这些设计简化了微芯片操作所需的入口数量，并创建了具有统一流动阻力的微通道，用于同步生成具有相同尺寸/体积的生物反应器。此外，该设计还包含一个机载梯度发生器，可以产生一系列药物浓度，并可以针对一个目标样本测试不同的治疗细胞系。单细胞分辨率和多种生物标志物的动态测量允许在治疗前后对任何细胞类型的特定状态进行动态评估。这使得激活和抑制的调节机制以及事件的因果关系得以确定。因此，拟议的芯片上方法可以同时评估疫苗的治疗潜力，关联机制和功能信息，从而在多个层面上更深入地了解细胞功能和反应。拟议研究开发的方法将进一步使我们能够回答重要的生物学问题。其中包括调节细胞疗法的随机性，以及细胞对各种免疫调节刺激的反应和变异。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。