⁶⁴Cu Hypoxia Imaging Radiotracer Targeting the Human Copper Transporter

The detection and characterization of hypoxia through functional imaging represents a critical frontier in oncology, cardiology, and neurology. Despite extensive efforts, the clinical translation of a robust hypoxia imaging biomarker remains hindered by suboptimal signal limiting diagnostic accuracy. Here, we report the development of a ⁶⁴Cu-based radiotracer (⁶⁴Cu-CysPhe), which exploits the cellular copper uptake pathway involving the human copper transporter 1 (hCtr1). This targeted mechanism facilitates selective tracer accumulation in hypoxic tissues. In-vivo PET-MRI imaging in a murine breast cancer model demonstrated tumor-to-muscle ratios exceeding 3.0 up to 24 h post-injection. Notably, immunohistochemical analyses revealed that ⁶⁴Cu-CysPhe preferentially localizes to hypoxic tumor regions while excluding necrotic cores, thereby providing information about a tumor's immediate microenvironment. Comparative analysis with established radiotracers, including ⁶⁴Cu-ATSM and ¹⁸F-FDG, confirmed the sensitivity of ⁶⁴Cu-CysPhe in detecting hypoxic tumor regions. These findings establish ⁶⁴Cu-CysPhe as a promising candidate for non-invasive imaging of tumors.