Endosome-anchored NLRP3 inflammasome
What we have shown so far
- Activation switch: Protein kinase D–dependent phosphorylation of NLRP3 at Ser295 acts as an activating switch that helps NLRP3 reach a competent assembly state.
- Endosomal recruitment: Upon activation, NLRP3 is recruited to endosomes.
- Permissive membrane: PI4P-enriched endosomal membranes provide the permissive platform for ASC filament nucleation and downstream signaling.
- Sequence of events: Endosomal recruitment → ASC polymerization → caspase-1 activation → cytokine release; supported by endogenous readouts in cells and tissues.
Project
Aim 1: Determinants of endosomal recruitment
Define the minimal membrane signature and thresholds for NLRP3 binding and nucleation using quantitative membrane reconstitution and live-cell validation.
Aim 2: Assembly sequence and structure
Resolve intermediate states on endosomal membranes with correlative imaging, cryo-EM/ET and ASC co-assembly analysis to obtain a mechanistic timeline.
Aim 3: Physiology and intervention
Test endosome-centric mechanisms in primary cells and mouse models; develop readouts to identify perturbations that tune NLRP3 output.
Key tools (selected)
- Liposome-based membrane reconstitution
- Endogenous complex imaging in cells and mouse tissues
- Proteo-lipidomics and interactome mapping
- Cryo-EM and cryo-ET of assembly states
- Genome-scale perturbation discovery
Public overview. Operational protocols, constructs, and parameter values are intentionally not disclosed.