Fu Jia, Zhang Yanan, Lv Yifan, Li Ruilin, Gu Hongchen, Yang Jingxing
School of Biomedical Engineering, Med-X Research Institute, Shanghai Jiao Tong University, Shanghai 200030, China.
Hefei Institute of Innovative Early Cancer Screening Technologies, Shanghai Jiao Tong University, Hefei 230000, China.
Int J Mol Sci. 2025 Sep 20;26(18):9199. doi: 10.3390/ijms26189199.
Genetic engineering of macrophages, particularly for chimeric antigen receptor macrophage (CAR-M) therapy, holds great promise for immunotherapy, yet is constrained by the challenge of efficient gene delivery into primary macrophages, which are notoriously resistant to transfection. While conventional strategies focus on optimizing the physicochemical properties of lipid nanoparticles (LNP), they often fail to overcome the intrinsic biological barriers of these cells. Here, we introduced a "bioactive nanocarrier" paradigm, hypothesizing that incorporating a cellular modulator directly into LNP structure can synergistically overcome these barriers. We designed and synthesized a novel LNP by integrating the pro-inflammatory fatty acid, arachidonic acid (ARA), as a functional structural component (ARA-LNP). Systematic optimization of the ARA content and mRNA payload revealed a formulation that achieves high transfection efficiency (83.76%) in primary M2-polarized bone marrow-derived macrophages (BMDMs), a cell type that recapitulates pro-tumoral phenotype in the tumor microenvironment. Leveraging this advanced delivery platform, we successfully generated HER2-targeting CAR-M that demonstrated potent and specific phagocytic activity against HER2-expressing tumor cells in vitro. This work presents a powerful strategy where the nanocarrier itself transiently modulates the target cell state to enhance gene delivery, providing a new design principle for engineering macrophages and other hard-to-transfect immune cells for therapeutic applications.
巨噬细胞的基因工程，特别是嵌合抗原受体巨噬细胞（CAR-M）疗法，在免疫治疗方面具有巨大潜力，但由于难以将基因高效递送至原代巨噬细胞而受到限制，众所周知，原代巨噬细胞对转染具有抗性。虽然传统策略侧重于优化脂质纳米颗粒（LNP）的物理化学性质，但它们往往无法克服这些细胞固有的生物学障碍。在此，我们引入了一种“生物活性纳米载体”范式，假设将细胞调节剂直接整合到LNP结构中可以协同克服这些障碍。我们通过整合促炎脂肪酸花生四烯酸（ARA）作为功能结构成分，设计并合成了一种新型LNP（ARA-LNP）。对ARA含量和mRNA负载量的系统优化揭示了一种在原代M2极化骨髓来源巨噬细胞（BMDM）中实现高转染效率（83.76%）的配方，BMDM是一种在肿瘤微环境中重现促肿瘤表型的细胞类型。利用这一先进的递送平台，我们成功生成了靶向HER2的CAR-M，其在体外对表达HER2的肿瘤细胞表现出强大且特异性的吞噬活性。这项工作提出了一种强大的策略，即纳米载体本身可短暂调节靶细胞状态以增强基因递送，为工程化巨噬细胞和其他难以转染的免疫细胞用于治疗应用提供了一种新的设计原则。
