Gene expression changes were assessed in tumor cells treated with epigenetic inhibitors and after drug withdrawal

Sequential therapy is a promising strategy to enhance antitumor efficacy; however, the rational design of sequential drug treatments remains challenging. Here, we introduce an AI framework, AlphaTherapy, which can design sequential drug combinations for cancer treatment. Analysis of AlphaTherapy-predicted combinations and large-scale in vitro validation revealed that the first drug in effective anti-tumor combinations often targets transcription, particularly epigenetic inhibitors. To investigate how epigenetic drugs enhance the efficacy of subsequent treatments, RNA-seq was performed on tumor cells treated with epigenetic inhibitors followed by drug withdrawal to assess gene expression changes.