Multi-Omics Analysis Reveals the Unexpected Immune Regulatory Effects of Arsenene Nanosheets in Tumor Microenvironment

Arsenene, a two-dimensional (2D) monoelemental layered nanosheet composed of arsenic, was recently reported to feature outstanding anticancer activities. However, the specific biological mechanism of action remains unknown. In this work, we extensively analyzed the mechanism of arsenene in vivo and in vitro and discovered the unexpected immune regulatory capability of arsenene for the first time. Analysis of cell phenotypes in tumor microenvironment by single-cell RNA sequencing revealed that arsenene remodeled the tumor microenvironment by recruiting a high proportion of anticancer immune cells to eliminate the tumor. Mechanistically, arsenene significantly activated T cell receptor signaling pathways to produce antitumor immune cells while inhibiting DNA replication and TCA cycle pathways of tumor cells in vivo. Further proteomic analysis on tumor cells revealed that arsenene induced reactive oxygen species production and oxidative stress damage by targeting thioredoxin TXNL1. The overloaded reactive oxygen species (ROS) further triggered endoplasmic reticulum stress responses to release damage-associated molecular patterns (DAMPs) and "eat-me" signals from dying tumor cells, leading to the activation of antigen-presenting processes to induce the subsequent effector tumor-specific CD8+ T cell immune responses. This unexpected discovery indicated for the first time that 2D inorganic nanomaterials could effectively activate direct anticancer immune responses, suggesting arsenene as a promising candidate nanomedicine for future cancer immunotherapy.