Elucidating spatial and temporal dynamics of macrophage polarization using bioluminescence microscopy

项目摘要/摘要： 巨噬细胞极化参与正常生理和病理过程 流程。巨噬细胞近乎普遍的重要性导致了对 对基因组和蛋白质组进行功能分类。然而，要衡量它仍然是一个挑战。 巨噬细胞的实时极化，例如在评估生物材料状态的情况下 植入物或纳米药物输送系统在实体肿瘤中的疗效。我们的预赛 数据显示，生物发光显微镜能够提供有价值的蛋白质表达数据 对于巨噬细胞的极化结合了传统成像系统的细胞跟踪能力。 这项研究计划将解决目前巨噬细胞实时评估方面的差距 通过1)进一步发展测量巨噬细胞极化的技术 利用生物发光显微镜的动力学和2)评估细胞内和细胞外的 环境刺激对感知疾病患者巨噬细胞极化最重要 环境背景。为了实现这些目标，我们将使用体外培养系统和 生物发光成像。这项工作将确定测量巨噬细胞的新策略 并验证在疾病背景下分析两极分化的重要性。最终， 这项研究将为合成生物学的发展带来新的战略，增强 诊断性测试和开发新的治疗靶点。此外，知识和 所开发的工具可以应用于极化至关重要的其他细胞，如干细胞 分化、化学吸引和细胞代谢。

Project Summary/Abstract: Macrophage polarization is involved in the progression of normal physiological and pathological processes. The near universal importance of macrophage has led to widescale assessment of the genomic and proteomic to classify their function. However, it is still a challenge to measure macrophage polarization in real time such as in the context of assessing the status of a biomaterial implant or the efficacy of a nanoparticle drug delivery system in a solid tumor. Our preliminary data show that bioluminescence microscopy is able to deliver the protein expression data valuable for macrophage polarization combined the cell tracking capability of traditional imaging system. This research program will address the current gap in real-time assessment of macrophage polarization by 1) further developing the technology to measure macrophage polarization dynamics using bioluminescence microscopy and 2) assessing the intrinsic and extracellular environmental stimuli that are most important for sensing macrophage polarization in a diseased environment context. To accomplish these goals, we will employ ex-vivo culture systems and live bioluminescence imaging. This work will identify novel strategies for measuring macrophage polarization and validate the importance of analyzing polarization in a disease context. Ultimately, this research will lead to new strategies for synthetic biology development, enhancement of diagnostic testing and development of new therapeutic targets. In addition, the knowledge and tools developed can be applied to other cells where polarization is paramount such as stem cell differentiation, chemoattraction, and cell metabolism.