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

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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.