Chiral Gd‐DOTA‐Functionalized Polymer Nanoprobe with GGT‐Triggered Charge Reversal for Tumor‐Specific MRI/Fluorescence Imaging

The development of tumor-specific contrast agents with enhanced sensitivity is crucial for precise cancer diagnostics. Herein, GGTPro-A-Cy5 is presented, a γ-glutamyl transpeptidase (GGT)-responsive polymeric dual-modal probe that integrates chiral Gd-DOTA for MRI and Cy5 for fluorescence imaging (FI). This probe leverages GGT overexpression in tumors to enable charge reversal-driven accumulation, achieving selective activation in the tumor microenvironment. The chiral Gd-DOTA, characterized by exceptional water-exchange rate and stability, is ideally suited for polymeric incorporation, yielding an excellent T₁ relaxivity of 28.9 mM^-1s^-1 in vitro-substantially higher than Gd-DOTA's 3.19 mM^-1s^-1. In vivo MRI in 4T1 tumor-bearing mice showed a significant 400% signal enhancement, compared to 25% for Gd-DOTA, accompanied by prolonged tumor retention following injection. Complementary fluorescence imaging confirmed GGT-dependent activation and tumor-specific signal persistence. Biodistribution studies demonstrated enhanced tumor uptake, and histopathological and serum analyses confirmed its biocompatibility without acute or subacute toxicity. GGTPro-A-Cy5 represents a significant advancement in dual-modal imaging, offering precise tumor targeting, high sensitivity, and strong potential for clinical translation in cancer diagnostics.