Aminopeptidases (APs) in the renin-angiotensin system (RAS) and their activity balance play crucial roles in regulating vascular functions. Multiplexed analysis of RAS-related AP activities is useful for diagnosing diseases including cancer. Dynamic nuclear polarization-coupled magnetic resonance imaging (DNP-MRI) enables the simultaneous detection of multiple enzymatic activities in vivo. However, developing practical DNP-MRI probes, especially for multiplexed detection, remains challenging. Here, we report the design of DNP-MRI probes for the in vivo multiplexed analysis of AP activities. By integrating quantum mechanical calculations, organic synthesis, and physicochemical and biochemical evaluations, we developed a series of AP-responsive DNP-MRI probes with high enzymatic reactivities and distinguishable chemical shifts. Using these probes, we successfully detected and visualized multiple AP activities in vivo. Furthermore, we performed in vivo multiplexed analysis of RAS-related AP activities in tumor-bearing mice, demonstrating the potential of this approach for monitoring the efficacy of antiangiogenic cancer therapy and for the accurate discrimination of tumor types.
Yatabe et al. (Thu,) studied this question.