A DNAzyme-Driven DNA Nanotracker for Orthogonal Dual-Channel Imaging of an Oncogenic Signaling Axis and Its Pharmacological Modulation

Abstract Pathway-targeted therapy has emerged as a promising strategy in cancer treatment, yet its efficacy relies on the accurate analysis of key signaling axes in living cells. The miR-140/FEN1 signaling axis, which is balanced in normal hepatocytes but dysregulated in hepatocellular carcinoma, represents a potential target for cancer therapy. Here, we present a DNAzyme-driven DNA nanotracker (DDN) for the orthogonal imaging of this pathway in dual channels with high sensitivity. The nanotracker is constructed by assembling DNAzyme-powered DNA walkers, which are responsive to miR-140 and FEN1, onto gold nanoparticles, enabling simultaneous visualization of upstream miRNA expression and downstream enzymatic activity in living cells. Leveraging this platform, we further revealed the differential pharmacological modulations of the miR-140/FEN1 signaling axis by simvastatin and myricetin. In summary, the DDN serves not only as a sensitive molecular imaging probe but also as a versatile tool for mechanistic studies, offering advanced opportunities for pathway-guided drug discovery and precision therapeutics.