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

人体能够在没有药物辅助的情况下杀死癌细胞。这种情况经常发生，归功于作为免疫系统一部分的细胞。它们被称为自然杀伤(NK)细胞。它们与癌细胞相互作用的机制还不是很清楚。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)出现之前不会检测到它。在这一点上，大量的肿瘤细胞与大量的NK细胞相互作用。如果观察到NK细胞单独与单个癌细胞相互作用，就可以了解更多关于NK细胞的功能。该项目将设计一种设备，通过创建包含一个NK细胞和一个癌细胞的液滴来进行这些观察。该设备将被用来测量互动的几个方面。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的关系，以及在不同药物存在时的活性。如果这个项目成功，它可能导致癌症免疫治疗策略的戏剧性改进，导致癌症诊断后更长、更健康的生命。此外，该项目将支持高中生和大学生的研究体验。该项目还将培训社区大学教师使用的许多研究技术，极大地放大对发展一支高技能生物技术工作队伍的积极影响。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，以确保有足够数量的反应用于筛选，并进行并行设计，以实现多路复用能力。并行设备设计将能够在多个生物反应存储阵列中筛选不同的药物/浓度/细胞组合。这些设计简化了微芯片运行所需的入口数量，并创建了具有统一流动阻力的微通道，用于同步生成相同尺寸/体积的生物反应器。此外，该设计结合了一个板载梯度发生器，可以产生一系列药物浓度，并能够针对一个目标样本测试不同的治疗细胞系。单细胞分辨率和多重生物标志物的动态测量允许在治疗前、治疗期间和治疗后动态评估任何细胞类型的特定状态。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，建议的芯片上方法允许同时评估疫苗的治疗潜力，将机制和功能信息关联起来，以在多个水平上更好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。其中包括随机性在规范基于细胞的治疗中的贡献，以及细胞对广泛的免疫调节刺激的反应和变化。该奖项反映了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。

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.