Cysteine cathepsin B (CTS-B) is a crucial enzyme that
is overexpressed
in numerous malignancies and contributes to the invasion and metastasis
of cancer. Therefore, this study sets out to develop and evaluate
an activity-based multimodality theranostic agent targeting CTS-B
for cancer imaging and therapy. A CTS-B activity-based probe, BMX2,
was synthesized and labeled efficiently with 68Ga and 90Y to produce 68Ga-BMX2 for multimodality imaging
and 90Y-BMX2 for radiation therapy. The affinity and specificity
of BMX2 binding with the CTS-B enzyme were determined by fluorescent
western blots using recombined active human CTS-B enzyme (rh-CTS-B)
and four cancer cell lines including HeLa, HepG2, MCF7, and U87MG,
with CA074 as the CTS-B inhibitor for control. Confocal laser scanning
microscope imaging and cell uptake measurement were also performed.
Then, in vivo PET imaging and fluorescence imaging were acquired on
HeLa xenografts. Finally, the therapeutic effect of 90Y-BMX2
was tested. BMX2 could be specifically activated by rh-CTS-B and stably
bound to the enzyme. The binding of BMX2 with CTS-B is time-dependent
and enzyme concentration-dependent. Although CTS-B expression varied
between cell lines, all showed significant uptake of BMX2 and 68Ga-BMX2. In vivo optical and PET imaging showed a high tumor
uptake of BMX2 and 68Ga-BMX2 and accumulation for more
than 24 h. 90Y-BMX2 could significantly inhibit HeLa tumor
growth. The development of 68Ga/90Y-BMX2, a
radioactive and fluorescent dual modality theranostic agent, demonstrated
an effective theranostic approach for PET diagnostic imaging, fluorescence
imaging, and radionuclide therapy of cancers, which may have a potential
for clinical translation for cancer theranostics in the future.