# 14-3-3ε–dependent deubiquitination and translocation of NLRP3 activates the inflammasome during sepsis

## TL;DR

This study shows how 14-3-3ε helps activate the NLRP3 inflammasome in sepsis, offering a new potential treatment target.

## Contribution

The study identifies 14-3-3ε as a novel positive regulator of NLRP3 inflammasome activation through deubiquitination and translocation.

## Key findings

- 14-3-3ε binds to NLRP3 and promotes its K63 deubiquitination and translocation to MAMs.
- Blocking 14-3-3ε improves survival and reduces organ injury in septic mice.
- Plasma 14-3-3ε levels are elevated in sepsis patients and correlate with disease severity.

## Abstract

The activation of the NLRP3 inflammasome is a pivotal step in hyperinflammation in sepsis; however, the regulatory mechanisms underlying its activation are not fully understood. In this study, we found that 14-3-3ε facilitates NLRP3 inflammasome activation by enhancing NLRP3 K63 deubiquitination and promoting its translocation to the mitochondria-associated ER membranes (MAMs) for full activation. Mass spectrometry revealed that 14-3-3ε binds to NLRP3 in macrophages during sepsis. Plasma 14-3-3ε levels were elevated in patients with sepsis and were positively associated with disease severity. 14-3-3ε promoted NLRP3 inflammasome activation by facilitating NLRP3 aggregation and NLRP3–ASC assembly. The interaction between 14-3-3ε and NLRP3 was dependent on phosphorylation at the S194 site of NLRP3 NACHT domain. The NLRP3–14-3-3ε interaction promoted K63 deubiquitination and enhanced the translocation of NLRP3 to MAMs, which is necessary for full activation of NLRP3 inflammasome. Furthermore, macrophage-conditional KO of 14-3-3ε or treatment with BV02, a 14-3-3 inhibitor, improved the survival rate and alleviated organ injuries in septic mice. Taken together, our data indicate that 14-3-3ε functions as a positive regulator of the NLRP3 inflammasome and could be a target for sepsis treatment.

14-3-3ε promotes NLRP3 inflammasome activation via K63 deubiquitination and MAM translocation, serving as a potential therapeutic target in sepsis.