Sensing and modulating the chemokine environment with synthetic cells

项目摘要 趋化因子的梯度控制着多种细胞类型的生理转运，其中许多都是相同的 趋化因子导致多种疾病，包括癌症、动脉粥样硬化和神经退行性变。控制 趋化因子梯度也提供了新的机会来改善治疗细胞的靶向招募 再生医学网站。趋化因子及其受体在生物医学中的重要作用 推动了调节这些途径的新药物的持续开发。然而， 针对趋化因子信号转导的化合物的临床翻译仍然缓慢，部分原因是尚未解决 关于局部趋化因子梯度如何控制细胞迁移的基本问题，特别是在疾病中 正常组织结构丧失的组织。而趋化因子的局部梯度被认为是关键 细胞运动的决定因素，测量或操纵趋化因子环境的方法 紧邻单元格的情况仍然有限。这项提案的目标是开发和利用合成纤维 细胞作为一种合成生物学工具来操纵趋化因子环境，这将有助于解决局部、 邻近细胞的趋化因子梯度引导细胞的趋化。具体地说，我们将重点关注两种趋化因子 受体通路，CXCL12-CXCR4和CCL21-CCR7，与多种疾病的进展密切相关 常见疾病和基于细胞的再生疗法的有希望的靶点。合成细胞可以是 自下而上设计的具有特殊功能的从头开始，包括分泌趋化因子。合成的 细胞不是活的，不生长或分裂，在未来的活体应用中使用可能是一种更安全的选择。 利用工程哺乳动物和合成细胞方面的最新合成生物学进展，合成 将构建具有自定义输入-输出关系的单元格。我们将设计合成细胞来检测 特定趋化因子的局部浓度，允许实时监测趋化因子梯度。我们也 将设计合成细胞，当与活细胞直接相互作用时，通过分泌趋化因子来做出反应 破坏呈现给活细胞的趋化梯度。这些工具将使我们能够了解 局部浓度的趋化因子调节活细胞的信号和运动。拟议中的工作 包括两个特定的目标：1)开发能够报告局部趋化因子浓度的合成细胞； 2)开发可调节趋化因子环境以调节趋化作用的合成细胞。这个 拟议的研究具有重要意义，因为它将哺乳动物细胞中已建立的合成生物学概念应用于 合成细胞，填补了工具开发中的一个根本空白，该空白阻碍了对 在复杂环境中的趋化性。这项工作还将产生持久的影响，为 使用工程合成细胞操纵局部趋化因子谱的潜在新干预措施 针对多种疾病环境下的治疗，具有特异性和可控性。

Project Summary Gradients of chemokines control physiologic trafficking of multiple cell types, and many of these same chemokines drive multiple diseases, including cancer, atherosclerosis, and neurodegeneration. Controlling chemokine gradients also offers novel opportunities to improve recruitment of therapeutic cells to target sites for regenerative medicine. Critical functions of chemokines and chemokine receptors in biomedicine has motivated ongoing development of new pharmaceutical agents regulating these pathways. However, clinical translation of compounds targeting chemokine signaling remains slow, due in part to unresolved basic questions about how local gradients of chemokines control cell migration, particularly in diseased tissues with loss of normal tissue architecture. While local gradients of chemokines are recognized as key determinants of cell movement, methods to measure or manipulate the chemokine environment immediately adjacent to cells remain limited. The objective of this proposal is to develop and utilize synthetic cells as a synthetic biology tool to manipulate the chemokine environment that will help address how local, cell-adjacent chemokine gradients steer chemotaxis of cells. Specifically, we will focus on two chemokine receptor pathways, CXCL12-CXCR4 and CCL21-CCR7, strongly associated with progression of several common diseases and promising targets for cell-based regenerative therapies. Synthetic cells can be engineered de novo from the bottom-up with specialized functions, including chemokine secretion. Synthetic cells are not alive, do not grow or divide, and may be a safer alternative to use in future in vivo applications. Leveraging recent synthetic biology developments in engineered mammalian and synthetic cells, synthetic cells with custom input-output relationship will be constructed. We will engineer synthetic cells to detect local concentrations of a specific chemokine, allowing real-time monitoring of chemokine gradients. We also will design synthetic cells that upon direct interaction with a living cell, respond by secreting a chemokine to disrupt the chemotactic gradient presented to a living cell. These tools will enable us to understand how local concentrations of a chemokine regulate signaling and movement of living cells. The proposed work consists of two specific aims: 1) To develop synthetic cells that can report local chemokine concentrations; and 2) To develop synthetic cells that modulate the chemokine environment to regulate chemotaxis. The proposed research is significant as it applies established synthetic biology concepts in mammalian cells to synthetic cells, filling a fundamental gap in tool development that has prevented complete understanding of chemotaxis in complex environments. The work will also have a lasting impact that opens the door for potential new interventions using engineered synthetic cells to manipulate local chemokine profiles specifically and controllably for therapy in multiple disease settings.