PSMA RNA aptamer A10-3.2 conjugated with indocyanine green for rapid intraoperative imaging and effective mild photothermal therapy of prostate cancer.

In prostate cancer (PCa) precision therapy, several challenges remain; among them, the delayed onset of intraoperative fluorescence imaging and the absence of targeted temperature control for local hyperthermia restrict the flexible perioperative use of tracers and increase the risk of thermal damage to adjacent sensitive tissues. To address these issues, we developed A10-3.2-ICG, a prostate-specific membrane antigen (PSMA)-targeting RNA aptamer conjugated with indocyanine green (ICG) via amine coupling. The probe was validated by mass spectrometry and spectroscopic analysis. For intraoperative imaging, A10-3.2-ICG demonstrated enhanced fluorescence and photothermal conversion efficiency compared to free ICG, enabling subminute delineation of PSMA-positive tumors with an imaging onset faster than most currently available intraoperative imaging probes for PCa, and sustained retention for 3-4 h in xenografts. Moreover, the probe effectively distinguished cancerous, adjacent, and benign tissues in clinical frozen sections. For therapy, A10-3.2-ICG enabled dose-tunable mild photothermal therapy (mPTT; 42-45 °C), which achieved 68.64 ± 5.34% tumor inhibition after repeated treatment cycles without inducing significant toxicity. This strategy overcomes the limitations of conventional PTT by offering precise thermal control with enhanced safety. Knockdown of AC026401.3, a novel therapeutic lncRNA target identified in our previous study-via siRNA transfection significantly inhibited PCa cell proliferation, migration, and invasion, and synergized with A10-3.2-ICG-mediated mPTT to enhance apoptosis. It also reduced SLC7A11 expression, suggesting a disruption of redox homeostasis in cancer cells. Taken together, this integrated strategy combining molecular targeting, real-time imaging, and localized mPTT, with optional gene regulation, offers a promising theranostic approach for precision management of PCa.