Microenvironment‐Guided Evolution of ssDNA‐SWCNT Probes for Selective Recognition of Aggressive Prostate Cancer Phenotypes

Despite advances in prostate cancer detection, distinguishing indolent from aggressive phenotypes remains challenging. We report a microenvironment-guided strategy for evolving phenotype-specific molecular probes using single-stranded DNA-functionalized single-walled carbon nanotubes (ssDNA-SWCNTs). Our approach employs 3D tumor models that recapitulate complex cancer microenvironments, enabling identification of ssDNA sequences with differential binding properties. We developed two distinct probes for prostate cancer cells: PC3D2, which preferentially binds hypoxia-adapted stem-like cells associated with treatment resistance, and PC2D2, which shows enhanced binding to mesenchymal-like cells. These probes exhibit characteristic second near-infrared (NIR-II, 1000-1700 nm) fluorescence, enabling non-invasive detection of aggressive phenotypes in heterogeneous tumors using NIR-II optical imaging. We demonstrate their utility for selective drug delivery to prostate cancer spheroids, resulting in enhanced therapeutic efficacy. This platform represents a significant advancement in precision diagnostics and theranostics, potentially transforming prostate cancer management through phenotype-specific targeting. The methodology offers a generalizable approach for developing nanoprobes that recognize clinically relevant cancer phenotypes based on their unique microenvironmental signatures rather than individual biomarkers.