Macrophage-Fibroblast Communication in Cell Migration and Extracellular Matrix Remodeling

项目摘要 细胞-细胞信号传导在动态环境中维持稳态功能，该动态环境涉及细胞- 衍生的旁分泌因子、机械提示和不同的氧气水平。巨噬细胞和成纤维细胞是关键 存在于几乎所有哺乳动物组织中的细胞类型，其整合来自其环境的不同信号， 参与组织内环境的稳定。现有的实验模型还不能精确地控制细胞衍生的 因子和氧水平，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，在理解这些问题方面存在着严重的知识差距。 在复杂的微环境中控制细胞间信号传导的基本机制。我的研究小组 我将通过研究两个关键问题来解决这一知识缺口：（1）巨噬细胞迁移是如何调节的 成纤维细胞分泌的旁分泌和3D环境中的机械提示之间的相互作用？(2)怎么 低氧水平调节成纤维细胞活化、ECM重塑和巨噬细胞-成纤维细胞串扰？到 解决第一个问题，我们将整合细胞内信号生物传感器与一种新的微流体技术 来精确控制旁分泌因子和细胞产生的力量。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体 具有集成的成纤维细胞活化的基于成像的多重分析和 机械力第二个问题的结果将提供对生理学的机械见解。 多细胞氧传感和ECM重塑的过程。我们过去的研究和使用3D的初步结果 微流控装置展示了工程化组织微环境和控制细胞生长的可行性。 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， 研究组织微环境中细胞迁移和ECM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.