The State Key Laboratory of Pharmaceutical Biotechnology, Division of Immunology, Medical School, Nanjing University, Nanjing 210093, PR China.
MOE Key Laboratory of High Performance Polymer Materials and Technology, Department of Polymer Science & Engineering, College of Chemistry & Chemical Engineering, and Jiangsu Key Laboratory for Nanotechnology, Nanjing University , Nanjing, 210093, PR China.
Theranostics. 2018 Nov 29;8(22):6307-6321. doi: 10.7150/thno.29746. eCollection 2018.
Macrophages orchestrate inflammation and control the promotion or inhibition of tumors and metastasis. Ferumoxytol (FMT), a clinically approved iron oxide nanoparticle, possesses anti-tumor therapeutic potential by inducing pro-inflammatory macrophage polarization. Toll-like receptor 3 (TLR3) activation also potently enhances the anti-tumor response of immune cells. Herein, the anti-tumor potential of macrophages harnessed by FMT combined with the TLR3 agonist, poly (I:C) (PIC), and FP-NPs (nanoparticles composed of amino-modified FMT (FMT-NH) surface functionalized with PIC) was explored. Proliferation of B16F10 cells co-cultured with macrophages was measured using immunofluorescence or flow cytometry (FCM). Phagocytosis was analyzed using FCM and fluorescence imaging. FP-NPs were prepared through electrostatic interactions and their properties were characterized using dynamic light scattering, transmission electron microscopy, and gel retardation assay. Anti-tumor and anti-metastasis effects were evaluated in B16F10 tumor-bearing mice, and tumor-infiltrating immunocytes were detected by immunofluorescence staining and FCM. FMT, PIC, or the combination of both hardly impaired B16F10 cell viability. However, FMT combined with PIC synergistically inhibited their proliferation by shifting macrophages to a tumoricidal phenotype with upregulated TNF-α and iNOS, increased NO secretion and augmented phagocytosis induced by NOX2-derived ROS . Combined treatment with FMT/PIC and FMT-NH/PIC respectively resulted in primary melanoma regression and alleviated pulmonary metastasis with elevated pro-inflammatory macrophage infiltration and upregulation of pro-inflammatory genes . In comparison, FP-NPs with properties of internalization by macrophages and accumulation in the lung produced a more pronounced anti-metastatic effect accompanied with decreased myeloid-derived suppressor cells, and tumor-associated macrophages shifted to M1 phenotype. mechanistic studies revealed that FP-NPs nanoparticles barely affected B16F10 cell viability, but specifically retarded their growth by steering macrophages to M1 phenotype through NF-κB signaling. FMT synergized with the TLR3 agonist PIC either in combination or as a nano-composition to induce macrophage activation for primary and metastatic melanoma regression, and the nano-composition of FP-NPs exhibited a more superior anti-metastatic efficacy.
巨噬细胞可以协调炎症反应，并控制肿瘤的促进或抑制以及转移。Ferumoxytol（FMT）是一种临床批准的氧化铁纳米颗粒，通过诱导促炎型巨噬细胞极化具有抗肿瘤治疗潜力。Toll 样受体 3（TLR3）的激活也能强烈增强免疫细胞的抗肿瘤反应。在此，研究人员探索了 FMT 联合 TLR3 激动剂聚（I：C）（PIC）和 FP-NPs（由氨基修饰的 FMT（FMT-NH）表面功能化的 PIC 组成的纳米颗粒）对巨噬细胞的抗肿瘤潜力。通过免疫荧光或流式细胞术（FCM）测量与巨噬细胞共培养的 B16F10 细胞的增殖。通过 FCM 和荧光成像分析吞噬作用。通过静电相互作用制备 FP-NPs，并通过动态光散射、透射电子显微镜和凝胶阻滞实验对其性质进行表征。在 B16F10 荷瘤小鼠中评估了抗肿瘤和抗转移作用，并通过免疫荧光染色和 FCM 检测肿瘤浸润免疫细胞。FMT、PIC 或两者的组合几乎不会损害 B16F10 细胞活力。然而，FMT 与 PIC 联合协同抑制其增殖，通过上调 TNF-α 和 iNOS，增加 NO 分泌并增强 NOX2 衍生的 ROS 诱导的吞噬作用，将巨噬细胞转变为杀瘤表型。FMT/PIC 和 FMT-NH/PIC 的联合治疗分别导致原发性黑色素瘤消退，并缓解肺部转移，同时增加促炎巨噬细胞浸润和上调促炎基因。相比之下，具有被巨噬细胞内化和在肺部积累特性的 FP-NPs 产生了更明显的抗转移作用，同时减少了骨髓源性抑制细胞，肿瘤相关巨噬细胞向 M1 表型转变。机制研究表明，FP-NPs 纳米颗粒几乎不影响 B16F10 细胞活力，但通过 NF-κB 信号通路将巨噬细胞转向 M1 表型，特异性地阻碍其生长。FMT 与 TLR3 激动剂 PIC 联合或作为纳米复合物协同诱导巨噬细胞活化，用于原发性和转移性黑色素瘤消退，FP-NPs 的纳米复合物表现出更优异的抗转移疗效。
