Particle-Assisted Control over Macrophage-Neutrophil interactions (Pac-Man)

巨噬细胞-中性粒细胞相互作用的粒子辅助控制（吃豆人）

• 批准号: 10725989
• 负责人: Kara Lorraine Spiller
• 金额: $ 20万
• 依托单位: DREXEL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10725989
• 关键词: Address; Adoptive Cell Transfers; Adoptive Transfer; Allogenic; Anti-Inflammatory Agents; Apoptotic; Autologous; Behavior; Biocompatible Materials; Cell Therapy; Cells; Cues; Data; Dexamethasone; Disease; Dose; Drug Delivery Systems; Fibrosis; Flow Cytometry; Gene Expression Profiling; Generations; Goals; Histology; Human; Immune; Immune response; In Situ; In Vitro; Inflammation; Inflammatory; Injury; Knowledge; Macrophage; Mechanics; Mediating; Mediator; Mus; Muscle; Patients; Pharmaceutical Preparations; Phenotype; Prevention; Process; Proliferating; Punch Biopsy; Regulatory T-Lymphocyte; Series; Site; Skeletal Muscle; Source; T-Cell Activation; T-Lymphocyte; Testing; Therapeutic; Time; Tissues; Trauma; biodegradable polymer; cell type; high reward; high risk; improved; in vitro testing; in vivo; in vivo Model; innovation; limb loss; manufacturing cost; mouse model; neutrophil; novel strategies; particle; prevent; quadriceps muscle; regenerative; repair model; repaired; response; satellite cell; tissue repair; uptake; volumetric muscle loss; wound healing

Project Summary Volumetric muscle loss (VML) is a debilitating injury caused by trauma or disease to skeletal muscle that leads to incapacitating fibrosis and loss of limb function. We and others have identified delayed clearance of apoptotic neutrophils as a primary mediator of fibrosis via detrimental effects on satellite cell and macrophage behavior. Because macrophages are critical regulators of wound healing and also the primary cell type that clear apoptotic neutrophils in a process called efferocytosis, macrophage cell therapy is a promising therapeutic approach, but is limited by two main challenges: 1) Macrophages are highly plastic cells that rapidly shift phenotype in response to microenvironmental cues. Therefore, a strategy is needed to control their phenotype in situ following administration, to prevent them from changing phenotype in response to pro-inflammatory cues at the site of injury. 2) High manufacturing costs and regulatory hurdles prevent the use of autologous (patient- derived) macrophages, especially because very high numbers are required, but allogeneic (donor-derived) macrophages elicit a strong T cell-mediated adverse immune response. We developed an innovative biomaterial-mediated macrophage cell therapy strategy that simultaneously addresses both of these challenges. In this strategy, referred to as Particle-Assisted Control over Macrophage-Neutrophil interactions (Pac-Man), the macrophages are first loaded ex vivo with polymeric biodegradable microparticles that slowly release dexamethasone intracellularly, thus controlling macrophage phenotype from the inside out following their administration to sites of injury. Dexamethasone was selected because it causes an anti-inflammatory/pro- regenerative phenotype in macrophages, increases their efferocytosis of apoptotic cells, and suppresses their ability to activate T cells. Thus, the intracellular release of dexamethasone following administration of the macrophages to sites of injury is expected to make them clear detrimental neutrophils, resolve inflammation, and suppress T cell activation to prevent rejection of allogeneic cells. In Aim 1, the functional phenotype of the adoptively transferred Pac-Man macrophages will be rigorously characterized over time in vitro and in vivo using flow cytometry, gene expression profiling, and analysis of their interactions with apoptotic neutrophils. Effects on muscle repair will be assessed using histology and functional mechanical testing. In Aim 2, the potential to use an allogeneic cell source will be tested in vitro and in vivo using primary human immune cells from mixed donors and mice from different strains. This project will advance an innovative off-the-shelf, translational cell therapy to enhance clearance of apoptotic cells, which would be transformative for the treatment of fibrotic injuries such as VML and many others.

项目摘要 肌肉体积损失（VML）是由骨骼肌的创伤或疾病引起的使人衰弱的损伤， 导致失能性纤维化和肢体功能丧失。我们和其他人已经确定， 凋亡中性粒细胞通过对卫星细胞和巨噬细胞的有害影响作为纤维化的主要介质 行为因为巨噬细胞是伤口愈合的关键调节器，也是清除伤口的主要细胞类型， 凋亡的中性粒细胞在一个过程中称为巨噬细胞增多症，巨噬细胞疗法是一种有前途的治疗方法 这种方法，但受到两个主要挑战的限制：1）巨噬细胞是高度可塑的细胞， 表型响应微环境线索。因此，需要一种策略来控制它们的表型 给药后原位，以防止它们响应促炎性信号而改变表型 在受伤的地方。2)高制造成本和监管障碍阻碍了自体（患者- 来源）巨噬细胞，特别是因为需要非常高的数量，但同种异体（供体来源） 巨噬细胞引起强烈的T细胞介导的不良免疫应答。我们开发了一种创新的 生物材料介导的巨噬细胞治疗策略，同时解决这两个挑战。 在该策略中，称为对巨噬细胞-神经元相互作用的颗粒辅助控制（Pac-Man）， 巨噬细胞首先离体负载聚合物可生物降解的微粒 地塞米松细胞内，从而控制巨噬细胞表型从内到外， 给药至损伤部位。选择地塞米松是因为它会产生抗炎/促炎作用 巨噬细胞的再生表型，增加它们对凋亡细胞的吞噬作用，并抑制它们的 激活T细胞的能力。因此，在施用本发明的化合物后，地塞米松的细胞内释放增加。 巨噬细胞的损伤部位，预计使他们清除有害的中性粒细胞，解决炎症， 抑制T细胞活化以防止同种异体细胞排斥。在目标1中， 过继转移的Pac-Man巨噬细胞将在体外和体内使用 流式细胞术、基因表达谱分析以及它们与凋亡中性粒细胞的相互作用的分析。影响 将使用组织学和功能性机械测试来评估肌肉修复。在目标2中，利用 将使用来自混合供体的原代人免疫细胞在体外和体内测试同种异体细胞来源 和不同品系的小鼠。该项目将推进一种创新的现成的转化细胞疗法， 增强凋亡细胞的清除，这对于治疗纤维化损伤， VML和许多其他人。