T-2 Toxin Induces Macrophage Extracellular Trap Formation via CLOCK-WNT10B-PAD2/ROS Signaling

Area of Science:

• Immunotoxicology
• Cellular Biology
• Molecular Mechanisms

Background:

• T-2 toxin is a potent immunotoxic mycotoxin.
• Mechanisms of T-2 toxin-induced macrophage extracellular trap (MET) formation are not fully understood.
• The role of the circadian regulator CLOCK in this process is unexplored.

Purpose of the Study:

• To investigate the kinetic features and molecular regulation of T-2 toxin-induced MET formation.
• To elucidate the involvement of the circadian regulator CLOCK in T-2 toxin-induced METs.
• To identify key molecular pathways driving T-2 toxin-induced METs.

Main Methods:

• Utilized RAW264.7 and THP-1 macrophage cell lines.
• Analyzed T-2 toxin-induced MET formation kinetics (14-32 nM, 24 h).
• Investigated the roles of PAD2, ROS, CLOCK, and WNT10B pathways.

Main Results:

• T-2 toxin induced METs in a time-dependent manner, with critical windows at 4 and 8 hours.
• The PAD2 pathway was identified as a continuous driver, with ROS playing a minor role.
• T-2 toxin suppressed CLOCK expression, relieving WNT10B repression and activating downstream pathways promoting MET formation.
• *Clock* knockdown enhanced MET release, confirming CLOCK's negative regulatory role.

Conclusions:

• T-2 toxin suppresses CLOCK expression, leading to WNT10B derepression and subsequent MET generation.
• This study reveals a novel mechanism of T-2 toxin-induced MET formation involving circadian clock regulation.
• Findings advance the understanding of mycotoxin immunotoxicity and macrophage extracellular trap formation.