Chemodynamic therapy (CDT) that kills tumor cells by converting low-reactivity H2O2 into highly toxic hydroxyl radicals (•OH) is an emerging tumor therapeutic modality, but its therapeutic efficacy is largely limited by both the lack of tumor targeting and redox homeostasis in tumor cells. Herein, we report Cu2+-encapsulated and GalNAc-imprinted biodegradable silica nanoparticles (nanoMIP) for boosting CDT. In this strategy, the Cu2+ was first encapsulated into disulfide-bridged silica nanoparticles with a high loading capacity of ∼18.3%, followed by in situ functionalization via molecular imprinting using GalNAc as a template. Such a nanovector could specifically target tumor cells overexpressing the Tn antigen to promote the cellular uptake. After internalization into tumor cells, the degradation of nanoMIP occurred in response to the tumor microenvironment, spontaneously releasing Cu2+/Cu+ via redox cycles, which in turn promoted highly potent GSH depletion and triggered •OH generation by a Fenton-like reaction. Notably, we found that the catalase activity could be effectively inhibited by the produced Cu+, which indirectly upregulated the endogenous H2O2 level. As a result, the “maladjusted” tumor cells lost the resistance against •OH damage, finally resulting in the apoptosis of tumor cells. In vitro and in vivo experiments demonstrated that our nanoMIP exhibited excellent cytotoxicity against tumor cells and high efficacy of tumor inhibition in the xenograft tumor model with negligible side effects. Taken together, our study provides not only a promising strategy for maximizing the CDT efficacy but also a new insight for developing MIP-based nanomedicine.

A sound understanding of cell migration behaviors and the internal mechanism is crucial for studying cancer metastasis and invasion. Continuous cell tracking and quantifying cellular and molecular dynamics of cell migration at the single-cell level is essential to elucidate rare, dynamic, and heterogeneous cell responses. Yet, a competent comprehensive analytical platform is lacking. Herein, we present an integrated single living cell analysis platform that enables long-term observation of migration behavioral phenotypes in single cells and simultaneous analysis of the signaling proteins and complexes during cell migration. Considering correlation between pathways and phenotypes, this platform is capable of analyzing multiple phenotypes and signaling protein dynamics at a subcellular resolution, reflecting the molecular mechanism of biological behavior. Using the EGFR-PI3K signaling pathway as a proof-of-principle, we explored how the pathway and related regulators, Rho GTPases, promote different migration phenotypes. Signaling pathway protein complexes p85α-p110α and p85α-PTEN were found to reciprocally modulate each other and subsequently regulate the expression level of the small GTPases by EGFR-related signaling pathways, which governs the cell migratory behavior. Thus, this single-cell analysis platform is a promising tool for rapid molecular mechanism analysis and direct observation of migration phenotypes at the single-cell level, providing insights into the molecular mechanism and phenotypes in cell migration.