An innovative NRF2 nano-modulator induces lung cancer ferroptosis and elicits an immunostimulatory tumor microenvironment

Abstract

Simultaneous targeting of both the tumor microenvironment and cancer cells by a single nanomedicine has not been reported to date. Here, we report the dual properties of zero-valent-iron nanoparticle (ZVI-NP) to induce cancer-specific cytotoxicity and anti-cancer immunity. Methods: Cancer-specific cytotoxicity induced by ZVI-NP was determined by MTT assay. Mitochondria functional assay, immunofluorescence staining, Western blot, RT-qPCR, and ChIP-qPCR assays were used to dissect the mechanism underlying ZVI-NP-induced ferroptotic cancer cell death. The therapeutic potential of ZVI-NP was evaluated in immunocompetent mice and humanized mice. Immune cell profiles of allografts and ex vivo cultured immune cells were examined by flow cytometry analysis, RT-qPCR assay, and immunofluorescence. Results: ZVI-NP caused mitochondria dysfunction, intracellular oxidative stress, and lipid peroxidation, leading to ferroptotic death of lung cancer cells. Degradation of NRF2 by GSK3/β-TrCP through AMPK/mTOR activation was enhanced in such cancer-specific ferroptosis. In addition, ZVI-NP attenuated self-renewal ability of cancer and downregulated angiogenesis-related genes. Importantly, ZVI-NP augmented anti-tumor immunity by shifting pro-tumor M2 macrophages to anti-tumor M1, decreasing the population of regulatory T cells, downregulating PD-1 and CTLA4 in CD8 + T cells to potentiate their cytolytic activity against cancer cells, while attenuating PD-L1 expression in cancer cells in vitro and in tumor-bearing immunocompetent mice. In particular, ZVI-NPs preferentially accumulated in tumor and lung tissues, leading to prominent suppression of tumor growth and metastasis. Conclusions: This dual-functional nanomedicine established an effective strategy to synergistically induce ferroptotic cancer cell death and reprogram the immunosuppressive microenvironment, which highlights the potential of ZVI-NP as an advanced integrated anti-cancer strategy.

Prognostic and Immunological Role of Key Genes of Ferroptosis in Pan-Cancer

“…Interestingly, our study also indicated that GPX4 expression was positively correlated with the expression levels of monocyte markers including CD14 and CD115 and M2 macrophage markers including VSIG4 and MS4A4A both in ESCA and in HNSC. Hsieh et al reported that zero-valent-iron nanoparticle (ZVI-NP) could promote the shift from pro-tumor M2 macrophages to anti-tumor M1 macrophages and finally inhibit the tumor progression ( Hsieh et al, 2021 ). Our results suggested that activation of M2 macrophages in ESCA and HNSC might be correlated with high expression of GPX4.…”

Targeting ferroptosis-based cancer therapy using nanomaterials: strategies and applications

“…Thus, the research to date has revealed that these nanomaterials significantly transformed TME macrophages from M2 to M1 phenotype, while having a good synergistic effect with anti-PD-L1 therapy 67 . Coincidentally, another study 68 has also reported that nanomaterials targeting ferroptosis can also promote the conversion of M2 macrophaes to M1 macrophages within the TME and attenuate PD-L1 expression in the TME. Their study suggests that ferroptosis promotes antitumor immunity 68 .…”

“…Coincidentally, another study 68 has also reported that nanomaterials targeting ferroptosis can also promote the conversion of M2 macrophaes to M1 macrophages within the TME and attenuate PD-L1 expression in the TME. Their study suggests that ferroptosis promotes antitumor immunity 68 . Another study, confirmed the promotion of the recruitment of infiltrating T cells by targeted ferroptosis nanomaterials and suggested their joint promotion with immunotherapy 69 .…”

Nanoparticles for Ferroptosis Therapy in Cancer

“…However, the polarization of TAMs toward the M1 tumorkilling type is attenuated by the release of anti-inflammatory factors as well as by altered metabolic functions of the macrophages. Among the strategies considered to promote the activation of TAMs, the use of NPs inducing FERR proved efficacy [59][60][61].…”

“…The simultaneous targeting of cancer cells and tumor microenvironment was achieved using zero-valent-iron NPs (ZVI-NP, Table 3). Aside from hitting cancer cells by inducing iron-mediated FERR, these NPs stimulate anticancer immunity by reprogramming the microenvironment [60]. Human (A549, H460, H1299) and mouse (LLC) lung carcinoma cells exposed to ZVI-NP showed alterations in redox balance, mitochondria dysfunction, and lipid peroxidation.…”