Macrophage-Fibroblast Communication in Cell Migration and Extracellular Matrix Remodeling

项目摘要 细胞-细胞信号在涉及细胞-细胞-的动态环境中维持动态平衡功能 派生的旁分泌因素，机械信号，和不同的氧气水平。巨噬细胞和成纤维细胞是关键 细胞类型存在于几乎所有哺乳动物组织中，它们整合了来自环境的不同信号，并 参与组织的动态平衡。现有的实验模型还不能精确地控制细胞来源 同时监测细胞迁移和细胞间通讯 细胞外基质(ECM)中的单个细胞水平。因此，关键的知识差距存在于理解 在复杂微环境中控制细胞间信号传递的基本机制。我的研究小组 将通过调查两个关键问题来解决这一知识差距：(1)巨噬细胞迁移是如何调节的 通过3D环境中成纤维细胞分泌的旁分泌和机械线索之间的相互作用？(2)如何 低氧水平调节成纤维细胞活化、细胞外基质重塑和巨噬细胞-成纤维细胞串扰？至 对于第一个问题，我们将把细胞内信号生物传感器与一种新的微流控技术相结合 精确控制旁分泌因子和细胞生成的力量。这些研究的结果将揭示 细胞迁移的基本原理。为了解决第二个问题，我们将设计多层微流控 具有基于集成成像的成纤维细胞激活和测量的多路分析设备 机械力。第二个问题的结果将提供对生理学的机械性见解 多细胞氧敏和细胞外基质重塑的过程。我们过去的研究和使用3D的初步结果 微流控设备展示了设计组织微环境和控制细胞的可行性 实时响应。总之，拟议的研究将建立一种新的基于微流体的方法来 研究组织微环境中细胞迁移和细胞外基质重塑的基本机制 时空定义的氧气景观、机械力和旁分泌因素。

Project Summary Cell-cell signaling maintains homeostatic functions in the presence of a dynamic environment that involves cell- derived paracrine factors, mechanical cues, and varying oxygen levels. Macrophages and fibroblasts are key cell types present in almost all mammalian tissues that integrate diverse signals from their environment and are involved in tissue homeostasis. Existing experimental models have not been able to precisely control cell-derived factors and oxygen levels while simultaneously monitoring cell migration and cell-cell communication at the single cell level in the extracellular matrix (ECM). Hence, a critical knowledge gap exists in understanding the fundamental mechanisms that control intercellular signaling in complex microenvironments. My research group will address this knowledge gap by investigating two key questions: (1) How is macrophage migration regulated by the interplay between fibroblast-secreted paracrine and mechanical cues in a 3D environment? (2) How do low oxygen levels modulate fibroblast activation, ECM remodeling, and macrophage-fibroblast crosstalk? To address the first question, we will integrate intracellular signaling biosensors with a novel microfluidic technology to control paracrine factors and cell-generated forces precisely. Results from these studies will uncover fundamental principles of cell migration. To address the second question, we will engineer multi-layer microfluidic devices with integrated imaging-based multiplexed analysis of fibroblast activation and measurement of mechanical forces. Results from the second question will provide mechanistic insights into the physiological process of multicellular oxygen-sensing and ECM remodeling. Our past studies and preliminary results using 3D microfluidic devices demonstrate the feasibility of engineering tissue microenvironments and controlling cellular responses in real-time. In summary, the proposed studies will establish a new microfluidics-based approach to studying basic mechanisms of cell migration and ECM remodeling in tissue microenvironments with spatiotemporally defined oxygen landscapes, mechanical forces, and paracrine factors.