Multigenerational lineage heterogeneity and metabolic plasticity of CD8 T cells

描述（由申请人提供）：从临床的角度来看，理解和操纵T细胞发育的动态可以为各种自身免疫性疾病的进展提供关键的见解，并为癌症免疫治疗和疫苗接种提供潜在的治疗选择。虽然现有的单细胞方法现在正在阐明异质性的重要性，但T细胞进化以实现特定表型状态的方式仍不清楚。我们建议使用新的微流体平台来动态监测几代人中单个CD8+ T细胞的反应，以研究分化，可塑性和代谢。为了实现这一点，我们将利用流体动力学陷阱，通过在芯片上实施标准免疫荧光技术来动态询问单个T细胞后代的几代。由于细胞周围的液体可以快速频繁地交换而不会干扰生长，因此我们将能够监测细胞表面标志物的表达，并在精确的时间点递送改变细胞代谢的药物。作为对谱系生化标志物的补充，我们还将测量单个活化T细胞的多代生长速率，以提供细胞代谢状态的物理测量。

DESCRIPTION (provided by applicant): From a clinical perspective, understanding and manipulating the dynamics of T cell development may offer key insight in to the progression of various autoimmune diseases as well as present potential treatment options with regards to cancer immunotherapy and vaccination. Although existing single cell approaches are now elucidating the significance of heterogeneity, the way in which T cells evolve to achieve particular phenotypic states remains unclear. We are proposing to use novel microfluidic platforms for dynamically monitoring the response of individual CD8+ T cells over several generations to investigate differentiation, plasticity and metabolism. To accomplish this, we will utilize hydrodynamic traps to dynamically interrogate several generations of a single T cell's progeny by implementing standard immunofluorescence techniques on-chip. Since fluid surrounding the cells can be rapidly and frequently exchanged without perturbing growth, we will be able to monitor expression of cell surface markers and deliver drugs that alter cellular metabolism at precise time points. As a compliment to biochemical markers of lineage we will also measure multigenerational growth rates of single activated T cells in order to provide a physical measurement of the cell's metabolic state.