Tumor lysate-cloaked CuMOF-sorafenib nanoassembler: A synergistic cuproptosis-ferroptosis nanoweapon against tumors

Abstract

Clinical tumor management is hindered by insufficient therapeutic efficacy and systemic side effects. Ferroptosis and cuproptosis, emerging metal ion-disruption-driven regulated cell death pathways, hold significant therapeutic potential. However, their combined efficacy is constrained by the tumor microenvironment (TME), where elevated levels of glutathione (GSH) and glutathione peroxidase 4 (GPX4) effectively mitigate reactive oxygen species (ROS) and lipid peroxidation. To overcome these barriers, we developed a biomimetic Cu-MOF-sorafenib nanoassembler (CMSP) for synergistic cuproptosis-ferroptosis-based anticancer therapy. CMSP comprises sorafenib (SOF)-loaded Cu-MOFs coated with tumor cell lysate-derived proteins. It exerts robust antitumor effects through three interconnected mechanisms: i) The tumor lysate coating facilitates homotypic targeting and immune evasion, thereby enhancing tumor-specific accumulation; ii) The acidic TME triggers the release of copper ions, which amplify ROS generation via Fenton-like reactions, while SOF inhibits System Xc^-, thereby depleting GSH and downregulating GPX4 expression; iii) Copper ions induce the aggregation of lipoylated proteins, initiating cuproptosis, while SOF synergistically promotes lipid peroxidation, driving ferroptosis. Both in vitro and in vivo studies demonstrate the efficient tumor accumulation, growth suppression, and metastasis inhibition of CMSP. By integrating biomimetic targeting, oxidative stress amplification, and dual-pathway synergy, CMSP presents a promising strategy to advance the development of personalized antitumor nanomedicines.

Keywords: Cuproptosis; Drug delivery system; Ferroptosis; Metal organic frameworks; Tumor lysate.

Graphical abstract

Scheme 1

Schematic illustration of the CMSP drug delivery system for synergistic cuproptosis-ferroptosis anticancer therapy. Schemes created with BioRender.com.

Fig. 1

Synthesis and characterization of CMSP. (a) SEM images of CM, CMS and CMSP. TEM image (b) and elemental mapping (c) of CMSP. (d) XRD pattern of CM, CMS and CMSP. (e) Zeta potential analysis of CM, CMS and CMSP. (f) XPS full spectrum of CMSP. (g) Valence analysis of Cu on the surface of CMSP. (h) Glutathione consumption curves of CM, CMS and CMSP. (i) UV–vis absorption and photo (inset) of TMB solution after different treatments (1: TMB; 2: TMB + GSH; 3: TMB + GSH + CMSP; 4: TMB + GSH + H₂O₂; 5: TMB + H₂O₂ + CMSP; 6: TMB + GSH + H₂O₂ + CM; 7: TMB + GSH + H₂O₂ + CMS; 8: TMB + GSH + H₂O₂ + CMSP). UV–vis absorption and photo (inset) of TMB solution after treatment at (j) different pH values and (k) different CMSP concentrations.

Fig. 2

CMSP induces 4T1 cell death and suppresses proliferation. (a) Toxicity assessment of CM, CMS, and CMSP at varying concentrations against 4T1 cells. (b) Live/dead staining of 4T1 cells following 24 h incubation with 100 μg/mL CM, CMS, and CMSP NPs. (c) Flow cytometry analysis of different death stages in 4T1 cells after 12 h incubation with 100 μg/mL CM, CMS, and CMSP NPs. (d) Representative images of 4T1 cell colony formation after 12 h incubation with CM, CMS, and CMSP. (e) Quantitative statistical analysis of the flow cytometry results shown in panel (c). (f) Flow cytometry analysis and (g) corresponding quantitative statistics of 4T1 cell proliferation using CFDA SE staining after 12 h incubation with CM, CMS, and CMSP. Data represent mean ± SD, n = 3, ns: not significant, ∗∗∗∗p < 0.0001.

Fig. 3

CMSP induces cellular damage by disrupting mitochondria function. (a) Fluorescence images of intracellular ROS content in 4T1 cells after incubation with CM, CMS and CMSP for 12 h. (b) Flow cytometry analysis and (c) corresponding quantitative statistics of intracellular ROS. (d) Fluorescence images of mitochondrial membrane potential after incubation of 4T1 with CM, CMS and CMSP for 12 h. JC-1 monomer (green) and JC-1 polymer (red). (e) Flow cytometry analysis and (f) corresponding quantitative statistics of mitochondrial membrane potential. Data represent mean ± SD, n = 3, ns: not significant, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗∗p < 0.0001. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Fig. 4

CMSP induce ferroptosis and cuproptosis. (a) Cell survival rate of 4T1 cells after being incubated with TTM, Ferr-1, CQ, TCEP and Z-FA-FMK for 6 h and CMSP for 24 h, respectively. (b) The GSH content in 4T1 cells after incubation with CM, CMS and CMSP for 12 h. (c) Fluorescence imaging and (d) flow cytometry analysis of the SLC7A11 protein expression in 4T1 cells after incubation with CM, CMS and CMSP for 12 h. (e) Flow cytometry analysis of FDX1 protein expression in 4T1 cells incubated with CM, CMS and CMSP for 12 h. (f) Fluorescence images of DLAT protein aggregation in 4T1 cells incubated with CM, CMS and CMSP for 12 h. (g) Western blot images of DLAT, GPX4 and FDX1. (h) Flow cytometry analysis and (i) fluorescence imaging of lipid ROS in 4T1 cells after incubation with CM, CMS and CMSP for 12 h. (j) The intracellular MDA content in 4T1 cells after incubation with CM, CMS and CMSP for 12 h. (k) Flow cytometry analysis and (l) fluorescence imaging of GPX4 protein expression in 4T1 cells incubated with CM, CMS and CMSP for 12 h. (m) TEM images of mitochondrial ultrastructure in 4T1 cells after different treatments. The red arrow indicates the mitochondria. Data represent mean ± SD, n = 3, ns: not significant, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Fig. 5

Antitumor efficiency in mice bearing 4T1 tumors treated with CM, CMS and CMSP. (a) Schematics depicting the therapeutic schedule in subcutaneous 4T1 cancer with intravenous injection of saline, CM, CMS or CMSP. Schemes created with BioRender.com. (b) Changes in the body weight of mice. (c) Changes in tumor volume. (d) Weight statistics of tumor tissues. (e) Images of tumor. (f–i) Tumor volume changes of saline, CM, CMS and CMSP treatment. (j) H&E and Ki67 staining of tumor tissue in saline, CM, CMS, and CMSP treatment groups. (k) TUNEL staining and (l) fluorescence statistics of tumor tissue in saline, CM, CMS, and CMSP treatment groups. (m) Images of India Ink staining in the lungs of mice implanted with tumor models and (n) results of lung metastases. Data represent mean ± SD, n = 3, ns: not significant, ∗p < 0.05, ∗∗∗∗p < 0.0001.

Fig. 6

CMSP induce ferroptosis and cuproptosis in mice bearing 4T1 tumors. Fluorescence staining of (a) DLAT, (b) FDX1, and fluorescence statistics of (c) DLAT, (d) FDX1; Fluorescence staining of (g) SLC7A11 and (h) GPX4 and fluorescence statistics of (e) SLC7A11 and (f) GPX4. (n = 3, ns: not significant, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001).