Multiplex Aptamer-Guided DNA Nanosuppressors for Quantitative Modulation of Autophagy–Survivin Crosstalk and Imaging-Guided Apoptosis Induction

Survivin, a critical inhibitor-of-apoptosis protein frequently overexpressed in cancer, represents a promising therapeutic target, yet its efficacy is confounded by the intricate interplay with autophagy. To quantitatively dissect this interplay, we engineer a panel of eight aptamer-guided gene suppressants (AGSs) on programmable DNA nanostructures, enabling simultaneous cancer-selective gene silencing and intracellular imaging. These AGSs specifically target Survivin, core autophagy-related genes (ATG5, ATG6, and ATG7), and additional oncogenes (PLK1, BMP-2, cMyc, and TK1). Systematic screening across 44 combination regimens reveals time- and magnitude-dependent rules governing autophagy modulation in Survivin-targeted therapy. Survivin knockdown alone induces hyperactivated autophagy (expression level ≥1.4), resulting in autophagic cell death. Conversely, concurrent or delayed autophagy inhibition sustains moderate autophagy levels (1.1-1.3), activating a cytoprotective, detoxification program. Most notably, autophagy preinhibition (<0.9) significantly sensitizes cells to subsequent Survivin silencing, markedly enhancing apoptosis. Furthermore, the AGS_Survivin platform incorporated a dual-input logic gate (nucleolin and Survivin), allowing selective fluorescence activation in targeted cancer cells (A549 and MCF-7) while sparing normal counterparts (MCF-10A). This work represents the first systematic application of multiplex AGSs to dissect Survivin-autophagy dynamics, establishing a quantitative correlation between autophagy expression and functional outcomes and demonstrating their potential as analytical tools for imaging-guided apoptosis induction.