Understanding and Controlling Macrophage Behavior in Angiogenesis

了解和控制血管生成中的巨噬细胞行为

• 批准号: 10629777
• 负责人: Kara Lorraine Spiller
• 金额: $ 7.74万
• 依托单位: DREXEL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-01-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10629777
• 关键词: Adoptive Transfer; Behavior; Biocompatible Materials; Cells; Cues; Data; Goals; Heart; Hindlimb; Hydrogels; In Vitro; Inflammatory; Inflammatory Response; Injury; Ischemia; Modeling; Mus; Phase; Phenotype; Population; Process; Site; T-Lymphocyte; Testing; Time; Tissues; Vascularization; Work; angiogenesis; bioscaffold; cell type; directed differentiation; healing; macrophage; monocyte; response; response to injury; scaffold; wound healing

Project summary At the heart of angiogenesis and biomaterial vascularization lies the inflammatory response, orchestrated primarily by macrophages, which dramatically shift phenotype over time in response to microenvironmental cues. In the normal response to injury, macrophages are initially pro-inflammatory (aka M1), and at later stages they are replaced by a mixed population referred to collectively as M2 that upregulate factors associated with resolution of the wound healing process. Previously we showed that M1 macrophages are critical for the initiation of angiogenesis, but they must switch to M2 for stable angiogenesis and wound healing. The extent of the diversity of this M2 population, and how they regulate angiogenic processes, is still unknown. At later stages of angiogenesis and biomaterial vascularization, M2 macrophages are generated 1) via transition from M1 macrophages, or 2) from direct differentiation of newly arriving monocytes. The differences between the M2 macrophages arising from each population have not been investigated. Preliminary data suggest that M1-derived M2 macrophages possess enhanced angiogenic functionality, and that biomaterials that transiently stimulate the initial M1 phase may enhance the subsequent response to M2-promoting biomaterials to achieve enhanced vascularization and healing. The overarching hypothesis of this project is that biomaterials that promote sequential M1 and M2 activation of the same population of macrophages will enhance vascularization. We recently found that adoptively transferred macrophages and M2-promoting microparticles are synergistic in promoting tissue revascularization in a murine hindlimb ischemia model. However, the macrophages were rapidly cleared from the site of injury (within 2 days), making it difficult to study the phenotype changes in adoptively transferred macrophages. Therefore, the goal of this project is to adoptively transfer macrophages using biomaterial scaffolds as a cell carrier, so that the macrophages are retained at the site of injury for long enough to track their phenotype changes and to observe their interactions with other cell types involved in angiogenesis. We will use porous scaffolds (as opposed to hydrogels) because other work in our lab has shown using in vitro studies that macrophage-T cell crosstalk is important for the M1-to-M2 transition of macrophages, so it is important that T cells are able to inflitrate the biomaterial carrier. In the current project, we will administer pro-inflammatory M1 macrophages to the site of injury using porous scaffolds in order to test the hypothesis that M1 macrophage crosstalk with T cells promotes Th2 differentiation in T cells, the M1-to-M2 transition in macrophages, and enhanced angiogenesis. The hypothesis is that adoptively transferred M1 macrophages will transition into a distinct M2 phenotype with enhanced angiogenic functions (compared to M0 macrophages) via crosstalk with endogenous T cells.

项目概要 血管生成和生物材料血管化的核心在于精心策划的炎症反应 主要是通过巨噬细胞，随着时间的推移，巨噬细胞会根据微环境线索显着改变表型。 在对损伤的正常反应中，巨噬细胞最初是促炎细胞（又名 M1），在后期阶段，它们 被统称为 M2 的混合群体所取代，M2 上调与 解决伤口愈合过程。之前我们表明 M1 巨噬细胞对于启动至关重要 血管生成，但它们必须切换到 M2 才能实现稳定的血管生成和伤口愈合。的程度 M2 群体的多样性以及它们如何调节血管生成过程仍然未知。在后期阶段 血管生成和生物材料血管化，M2 巨噬细胞通过 1) 从 M1 转变而产生 巨噬细胞，或 2) 来自新到达的单核细胞的直接分化。 M2与M2的区别 尚未对每个群体产生的巨噬细胞进行研究。初步数据表明 M1 衍生 M2 巨噬细胞具有增强的血管生成功能，并且瞬时刺激血管生成的生物材料 初始 M1 阶段可能会增强对 M2 促进生物材料的后续反应，以实现增强 血管化和愈合。该项目的总体假设是生物材料可以促进 同一巨噬细胞群的连续 M1 和 M2 激活将增强血管化。 我们最近发现过继转移的巨噬细胞和 M2 促进微粒 在小鼠后肢缺血模型中协同促进组织血运重建。然而， 巨噬细胞从损伤部位迅速清除（2天内），使得研究表型变得困难 过继转移的巨噬细胞的变化。因此，该项目的目标是过继转移 巨噬细胞利用生物材料支架作为细胞载体，使巨噬细胞保留在 损伤足够长的时间来跟踪它们的表型变化并观察它们与其他细胞类型的相互作用 参与血管生成。我们将使用多孔支架（而不是水凝胶），因为我们实验室的其他工作 体外研究表明，巨噬细胞-T 细胞串扰对于 M1 到 M2 的转变非常重要 巨噬细胞，因此 T 细胞能够浸润生物材料载体非常重要。在当前的项目中，我们 将使用多孔支架将促炎 M1 巨噬细胞注入损伤部位，以测试 假设 M1 巨噬细胞与 T 细胞的相互作用促进 T 细胞中的 Th2 分化，即 M1 到 M2 巨噬细胞的转变，并增强血管生成。 假设过继转移的 M1 巨噬细胞将转变为独特的 M2 巨噬细胞 通过与M0巨噬细胞的串扰增强血管生成功能的表型（与M0巨噬细胞相比） 内源性T细胞。