PROTAC Mediated Structure-Function Disruption of CD26 A Therapeutic Strategy for Driver Negative NSCLC

Background: Previously PROTAC research in NSCLC has focused on targeting driver mutations to overcome resistance, leaving 40% of patients lacking actionable alterations without therapeutic options. CD26-a transmembrane glycoprotein overexpressed in NSCLC with dual oncogenic functions-represents an unexplored PROTAC target for driver mutation-negative tumors.To explore the transcriptional changes induced by P43 in H460 non-small cell lung cancer cells, we performed bulk RNA sequencing (RNA-seq) on cells treated with DMSO (vehicle control) or P43 for 24 hours. Each condition included 6 biological replicates (total n=12 samples).Briefly, H460 cells were treated with either DMSO or P43 at the predetermined concentration for 24 hours under standard culture conditions. Total RNA was isolated from each sample, followed by DNase I treatment to eliminate genomic DNA contamination. RNA quality was assessed to ensure suitability for sequencing.Interpretation: Targeted CD26 degradation represents a novel therapeutic strategy exploiting protein overexpression rather than mutational dependency. By concurrently disrupting CD26s dual oncogenic roles, this approach not only showed efficacy in NSCLC in both in vivo and in vitro but also addresses limitations of mutation-targeted PROTAC based therapies, offering promise for driver mutation-negative NSCLC patients with high CD26 expression. P4-3s preclinical efficacy and safety profile support further clinical translation attempts while its mechanism provides a blueprint for targeting other membrane proteins.