Multiplexed Transcriptomics for Screening Drug Combinations and Defining the Mechanism of Action of HCC Therapeutics...

Area of Science:

• Oncology
• Genomics
• Pharmacology

Background:

• Hepatocellular carcinoma (HCC) remains a leading cause of cancer-related mortality worldwide.
• Classical drug screening methods lack the throughput and detailed cellular response information needed for complex cancer therapies.
• Single-cell sequencing technologies offer enhanced capabilities for drug discovery and understanding treatment mechanisms.

Purpose of the Study:

• To screen clinical drug combinations for anti-hepatocellular carcinoma (HCC) activity using a high-throughput single-nucleus sequencing platform.
• To elucidate the mechanism of action for effective drug combinations identified.
• To identify key regulators and cellular responses involved in drug-induced tumor suppression.

Main Methods:

• Utilized the snHH-seq platform for high-throughput screening of drug combinations against HCC.
• Performed single-cell transcriptomics analysis to identify effective drug combinations and their effects.
• Conducted in vitro and in vivo functional assays, ferroptosis assays, and gene regulatory network analysis.

Main Results:

• The HY combination (HHT and YM155) demonstrated the strongest suppression of HCC cell proliferation.
• HY treatment was found to induce ferroptosis in HCC cells, confirmed by rescue experiments.
• JUN was identified as a key regulator of proliferation inhibition, particularly in apoptotic tumor cell subclusters.

Conclusions:

• The HY drug combination is a promising therapeutic strategy for HCC, acting via ferroptosis induction.
• Integrating high-throughput single-cell screening with mechanistic studies accelerates the discovery of targeted combination therapies.
• This approach provides a framework for precise HCC interventions by exploiting pathway vulnerabilities and cellular heterogeneity.