In vivo imaging of early stage apoptosis by measuring real-time caspase-3/7 activation |
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Authors: | Matteo Scabini Fabio Stellari Paolo Cappella Sara Rizzitano Gemma Texido Enrico Pesenti |
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Institution: | (1) Pharmacology Department, Oncology, Nerviano Medical Sciences, Viale Pasteur 10, 20014 Nerviano, Italy;(2) Biology Department, Oncology, Nerviano Medical Sciences, Viale Pasteur 10, 20014 Nerviano, Italy; |
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Abstract: | In vivo imaging of apoptosis in a preclinical setting in anticancer drug development could provide remarkable advantages in
terms of translational medicine. So far, several imaging technologies with different probes have been used to achieve this
goal. Here we describe a bioluminescence imaging approach that uses a new formulation of Z-DEVD-aminoluciferin, a caspase
3/7 substrate, to monitor in vivo apoptosis in tumor cells engineered to express luciferase. Upon apoptosis induction, Z-DEVD-aminoluciferin
is cleaved by caspase 3/7 releasing aminoluciferin that is now free to react with luciferase generating measurable light.
Thus, the activation of caspase 3/7 can be measured by quantifying the bioluminescent signal. Using this approach, we have
been able to monitor caspase-3 activation and subsequent apoptosis induction after camptothecin and temozolomide treatment
on xenograft mouse models of colon cancer and glioblastoma, respectively. Treated mice showed more than 2-fold induction of
Z-DEVD-aminoluciferin luminescent signal when compared to the untreated group. Combining D-luciferin that measures the total tumor burden, with Z-DEVD-aminoluciferin that assesses apoptosis induction via caspase
activation, we confirmed that it is possible to follow non-invasively tumor growth inhibition and induction of apoptosis after
treatment in the same animal over time. Moreover, here we have proved that following early apoptosis induction by caspase
3 activation is a good biomarker that accurately predicts tumor growth inhibition by anti-cancer drugs in engineered colon
cancer and glioblastoma cell lines and in their respective mouse xenograft models. |
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