A Novel Bioluminescent Biosensor Quantifying Intramolecular Interaction and Levels of Pyroptosis Effector GSDMD

Area of Science:

• Biochemistry
• Molecular Biology
• Cell Biology

Background:

• Gasdermin D (GSDMD) executes pyroptosis, a crucial inflammatory programmed cell death pathway.
• GSDMD's role in diseases like heart failure and cancer highlights its therapeutic potential.
• Studying GSDMD function is difficult due to its lack of enzymatic activity.

Purpose of the Study:

• To develop a novel bioluminescent biosensor for Gasdermin D (GSDMD).
• To enable real-time monitoring of GSDMD dynamics and pyroptosis in living cells.
• To facilitate drug discovery for GSDMD-targeted therapies in inflammatory diseases.

Main Methods:

• Utilized NanoBiT technology to create a GSDMD bioluminescent biosensor (GSDMD-BS).
• Quantified intramolecular interactions, uncleaved GSDMD levels post-caspase-1 cleavage, and expression in live cells.
• Validated sensor accuracy in vitro and demonstrated in vivo potential using a xenograft mouse model.

Main Results:

• Successfully developed a functional GSDMD bioluminescent biosensor (GSDMD-BS).
• The biosensor accurately measures GSDMD levels and cleavage events in vitro.
• Demonstrated the biosensor's applicability in a preclinical mouse model.

Conclusions:

• The novel GSDMD biosensor provides a valuable tool for real-time pyroptosis monitoring.
• This technology will accelerate research into GSDMD and pyroptosis.
• The biosensor can be applied to high-throughput drug screening for inflammatory diseases.