M2 macrophage-targeted iron oxide nanoparticles for magnetic resonance image-guided magnetic hyperthermia therapy

doi:10.1016/j.jmst.2020.11.058

Abstract

Abstract:

Tumor-associated macrophages (TAMs) play an important role in tumor development and progression. In particular, M2 TAMs can promote tumor growth by facilitating tumor progression and malignant behaviors. Selectively targeted elimination of M2 TAMs to inhibit tumor progression is of great significance for cancer treatment. Iron oxide nanoparticles based magnetic hyperthermia therapy (MHT) is a classical approach to destroy tumor tissue with deep penetration depth. In this study, we developed a typical M2 macrophage-targeted peptide (M2pep) functionalized superparamagnetic iron oxide nanoparticle (SPIO) for magnetic resonance imaging (MRI)-guided MHT in an orthotopic breast cancer mouse model. The obtained multifunctional SPIO-M2pep with a hydrodynamic diameter of 20 nm showed efficient targeting capability, high transverse relaxivity (149 mM^-1 s^-1) and satisfactory magnetic hyperthermia performance in vitro. In vivo studies demonstrated that the SPIO-M2pep based MRI can monitor the distribution of nanoparticles in tumor and indicate the suitable timing for MHT. The M2 macrophage-targeted MHT significantly reduced the tumor volume and the population of pro-tumoral M2 TAMs in tumor. In addition, the SPIO-M2pep based MHT can remodel the tumor immune microenvironment (TIME). The multifunctional SPIO-M2pep with M2 macrophage-targeting ability, high magnetic hyperthermia efficiency, MR imaging capability and effective role in remodeling the TIME hold great potential to improve clinical cancer therapy outcomes.

Key words: M2 macrophages-targeted peptide, Iron oxide nanoparticles, Magnetic resonance imaging, Magnetic hyperthermia therapy, Tumor immune microenvironment

Wenshen Wang, Fenfen Li, Shibo Li, Yi Hu, Mengran Xu, Yuanyuan Zhang, Muhammad Imran Khan, Shaozhen Wang, Min Wu, Weiping Ding, Bensheng Qiu. M2 macrophage-targeted iron oxide nanoparticles for magnetic resonance image-guided magnetic hyperthermia therapy[J]. J. Mater. Sci. Technol., 2021, 81: 77-87.

Figures/Tables 8

Scheme. 1. Schematic of the preparation of SPIO-M2pep and the mechanism of SPIO-M2pep in M2 TAM-targeted and MRI-guided magnetic hyperthermia therapy of breast cancer.

Fig. 1. Characterization of SPIO and SPIO-M2pep nanoparticles. Transmission electron microscopy (TEM) images of (A) SPIO and (B) SPIO-M2pep nanoparticles with negative staining (inset: the size distribution analyses). (C) UV-vis absorption spectra. (D) Fluorescence emission spectra excited at a wavelength of 470 nm. (E) In vitro temperature increase profile of SPIO and SPIO-M2pep (6.0 μg μL -1) under an alternating magnetic field (2.56 kA m-1, 351 kHz). (F) The specific absorption rate values of SPIO (57.06 ± 0.78 W g -1) and SPIO-M2pep (56.03 ± 1.59 W g -1) under an AMF (2.56 kA m-1, 351 kHz).

Fig. 2. Cytotoxicity and targeting ability of SPIO-M2pep. (A) MTT analysis of SPIO or SPIO-M2pep on M2 macrophage cells. (B) MTT analysis of SPIO or SPIO-M2pep on 4T1 cells. (C) Prussian blue staining of M2 macrophage cells after different treatments. (D) Iron uptake in M2 macrophage cells treated with SPIO and SPIO-M2pep for different incubation times (25 μg mL -1, n = 3).

Fig. 3. In vitro magnetic hyperthermia therapy performance. (A) MTT analysis of M2 macrophage cells after treatment in a water bath for different times at 42 °C. (B) Fluorescence images of Hoechst/PI staining of M2 macrophage cells with different treatments under AMF (blue: Hoechst 33342 staining of nucleus, red: PI staining of dead cells, green: FITC signal from SPIO-M2pep). (C) Quantitative statistics of cell viability from cell fluorescence images after MHT treatment.

Fig. 4. MR imaging abilities of SPIO and SPIO-M2pep nanoparticles in vitro and in vivo. (A) T2-weighted MR images of SPIO and SPIO-M2pep nanoparticles dispersed in water at various concentrations of Fe. (B) Relaxivities (r2) of the SPIO and SPIO-M2pep nanoparticles measured at 3.0 T. (C) MR imaging of orthotopic breast tumor mice before and 30 min after injection of SPIO or SPIO-M2pep (tumor: yellow dashed line).

Fig. 5. M2 TAMs targeted MHT hinders cancer progression. (A) Infrared radiation (IR) thermal images of orthotopic breast tumor mice treated with MHT for 15 min. (B) Tumor growth curves in tumor-bearing mice treated with MHT after different treatment. (C) Tumor weights of different groups taken on day 14. (D) Representative pictures showing the lung metastases. E) The number of lung metastasis nodules of different groups (n = 5). (F) Body weight records of tumor-bearing mice with different treatment. (G) H&E analysis of the lung tissues collected from different groups.

Fig. 6. SPIO-M2pep-mediated MHT selectively reduced and repolarized M2 TAMs in tumor sites. (A) Representatives of the treatments of PBS, SPIO, and SPIO-M2pep in mice with or without AMF, depicting the F4/80 (APC) and CD206 (PE) expression on TAMs by flow cytometry. (B) Quantitative statistical analysis of F4/80+ and CD206+ macrophages from the flow cytometry results (mean ± SD; n = 3-4 per group). (C) Immunohistochemical staining of CD163+ macrophages from tumor tissues after treatment. (D) Quantitative statistical analysis of CD163+ macrophages from the immunohistochemistry imagies (mean ± SD; n = 5). (E) Immunofluorescence images of tumor sections stained with CD11b, CD86 and CD206.

Fig. 7. Remodeling of TIME. (A) CD3+ in CD45+ cells. (B) CD8+ in CD3+ cells. (C) CD4+ in CD3+ cells. (D) CD69+ in CD3+ cells. (E) CD69+ in CD8+ cells. (F) CD69+ in CD4+ cells. (G) IF assay of CD3, CD8, Granzyme B, TNF-α, IFN-γ, IL-10 and TGF-β.

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