Decrypting drug actions and protein modifications by dose- and time-resolved proteomics.

Area of Science:

• Proteomics
• Cancer Biology
• Pharmacology

Background:

• Most cancer drugs target cellular pathways by altering posttranslational modifications (PTMs).
• Limited understanding exists regarding the scope and dynamics of drug-induced PTM changes.
• Characterizing drug-PTM interactions is crucial for understanding drug efficacy and mechanisms.

Purpose of the Study:

• To develop and validate a proteomic assay, decryptM, for quantifying drug-modulated PTMs.
• To investigate drug target engagement and mechanism of action through PTM analysis.
• To create a comprehensive resource of drug-PTM modulation profiles.

Main Methods:

• Development of decryptM, a proteomic assay to measure thousands of PTMs in response to drug treatment.
• Application of decryptM to profile 31 cancer drugs across 13 cell lines.
• Generation of 1.8 million dose-response curves for drug-PTM interactions.

Main Results:

• DecryptM successfully quantifies drug-PTM modulation, providing insights into target engagement.
• Examples include detecting DNA damage from chemotherapeutics and identifying PTM signatures of kinase inhibitors.
• Rituximab's mechanism of killing CD20-positive B cells via B cell receptor overactivation was demonstrated.

Conclusions:

• DecryptM is a broadly applicable proteomic approach for dissecting drug mechanisms of action.
• The assay provides valuable data on drug-specific PTM responses.
• The generated dataset serves as an interactive molecular resource for cancer drug research.