Dual-target regulation of prostate cancer genetic dependencies by a programmable tetrahedral DNA nanostructure

Prostate cancer can depend on interconnected transcriptional regulators and functionally redundant paralogs, both of which may limit the effectiveness of single-target intervention. To enable coordinated dual-target regulation, we developed TetraArm-TDN, a programmable tetrahedral DNA nanostructure that integrates two antisense specificities within a single scaffold. We examined two biologically distinct target pairs: AR/FOXA1, which cooperate in prostate lineage transcription, and SMARCA2/SMARCA4, the alternative catalytic ATPases of SWI/SNF complexes. AR/FOXA1 taTDN concurrently reduced both proteins, attenuated androgen-responsive transcription, and impaired cell-cycle progression with G1-phase accumulation. Coordinated SMARCA2/SMARCA4 suppression produced stronger inhibition of prostate cancer cell growth than single-target treatments and was associated with increased cleaved PARP. In 22Rv1 xenografts, AR/FOXA1 dual targeting inhibited tumor growth both alone and in combination with enzalutamide. Mechanistically, AR and FOXA1 protein reduction occurred without proportional depletion of their corresponding mRNAs. Among the FOXA1-targeting constructs tested, the CDS-directed construct produced stronger protein suppression than the UTR-directed constructs, while polysome profiling showed reduced FOXA1 mRNA association with heavier polysome fractions. Together, these findings show that TetraArm-TDN can coordinately regulate transcriptionally cooperative and paralog-redundant targets in prostate cancer and suggest that translation-associated regulation may contribute to its target-suppressive activity.