Exome Sequencing of Drug-Resistant Clones for Target Identification

Area of Science:

• Oncology
• Molecular Biology
• Genomics

Background:

• Phenotype-based screening identifies small molecules with anticancer activity.
• Determining the specific protein targets of these compounds remains a significant challenge in drug discovery.
• Current gold standard methods for target identification involve genetic validation in vitro and in vivo.

Purpose of the Study:

• To establish a robust procedure for identifying the molecular targets of small molecule anticancer compounds.
• To enable rapid target discovery for cytotoxic agents using a genetic approach.

Main Methods:

• Utilized the colorectal cancer cell line HCT-116 to generate drug-resistant clones.
• Employed whole-exome sequencing to identify recurrent mutations in resistant cells.
• Integrated downstream in vitro and in vivo validation experiments to confirm target engagement.

Main Results:

• Successfully isolated drug-resistant clones from HCT-116 cells.
• Identified recurrent mutations associated with compound resistance through whole-exome sequencing.
• Demonstrated the feasibility of this strategy for identifying drug targets.

Conclusions:

• This combined approach of generating resistant clones and whole-exome sequencing provides a powerful tool for rapid small molecule target identification.
• The method facilitates the discovery of novel therapeutic targets for anticancer drug development.