Profiling of MicroRNA Targets Using Activity-Based Protein Profiling: Linking Enzyme Activity to MicroRNA-185...

Area of Science:

• Biochemistry
• Molecular Biology
• Immunometabolism

Background:

• MicroRNAs (miRNAs) regulate cellular processes by repressing protein translation.
• Traditional methods often fail to fully elucidate global miRNA functions.
• Understanding miRNA-185's role in immunometabolism requires advanced profiling.

Purpose of the Study:

• To investigate the functional alterations caused by microRNA-185 (miR-185) in human liver cells.
• To identify specific protein targets and metabolic pathways regulated by miR-185.
• To explore the potential for pharmacological intervention based on miR-185 activity.

Main Methods:

• Activity-based protein profiling using fluorophosphonate probes.
• Transcriptomics and lipidomics analyses.
• Functional assays involving monoglyceride lipase inhibition.

Main Results:

• miR-185 specifically altered the activity of metabolic serine hydrolase enzymes.
• Enzymatic changes correlated with alterations in cellular lipid species, including very-low-density lipoprotein components.
• Inhibition of monoglyceride lipase, a miR-185 target, reduced hepatitis C virus levels.

Conclusions:

• The study identified key functional changes in serine hydrolases regulated by miR-185.
• These miR-185-mediated changes in serine hydrolase activity are pharmacologically targetable.
• A small molecule inhibitor demonstrated the potential to recapitulate the phenotypic effects of miR-185.