A screen for MeCP2-TBL1 interaction inhibitors using a luminescence-based assay

Area of Science:

• Neuroscience
• Molecular Biology
• Genetics

Background:

• MeCP2 duplication syndrome (MDS) is a severe neurodevelopmental disorder characterized by neuronal dysfunction due to elevated MeCP2 levels.
• MeCP2 interacts with the nuclear co-repressor (NCoR) complex via TBL1 and TBLR1, a binding site crucial for MeCP2 toxicity in MDS models.

Purpose of the Study:

• To develop a high-throughput screening assay to identify small molecules that disrupt the MeCP2-TBL1/TBLR1 interaction.
• To facilitate the discovery of potential therapeutics for MeCP2 duplication syndrome.

Main Methods:

• A NanoLuc luciferase complementation assay was designed to measure the MeCP2-TBL1/TBLR1 interaction.
• A dual screening approach was employed, combining the primary assay with a counter-screen using protein kinase A (PKA) subunits.
• Compound libraries were interrogated using this dual screening strategy.

Main Results:

• The NanoLuc assay demonstrated high performance with excellent control separation and low signal variance (Z-factor = 0.85).
• The dual screening approach successfully identified candidate inhibitors of the MeCP2-TBL1/TBLR1 interaction.
• The study validated the feasibility of screening large compound collections for therapeutic leads.

Conclusions:

• The developed assay is a viable tool for identifying small molecules targeting the MeCP2-TBL1/TBLR1 interaction.
• This work paves the way for developing novel small molecule therapeutics to treat MeCP2 duplication syndrome.
• Future large-scale screens are anticipated to yield promising drug candidates for MDS.