title: In vivo imaging of early stage apoptosis by measuring real-time caspase-3/7 activation.
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.
authors.affiliation: Pharmacology Department, Oncology, Nerviano Medical Sciences, Viale Pasteur 10, 20014 Nerviano, Italy.
authors.forename: Matteo
authors.initials: M
authors.lastname: Scabini
authors.forename: Fabio
authors.initials: F
authors.lastname: Stellari
authors.forename: Paolo
authors.initials: P
authors.lastname: Cappella
authors.forename: Sara
authors.initials: S
authors.lastname: Rizzitano
authors.forename: Gemma
authors.initials: G
authors.lastname: Texido
authors.forename: Enrico
authors.initials: E
authors.lastname: Pesenti
mesh_terms: D000818:Animals; D017209:Apoptosis; D015153:Blotting, Western; D002166:Camptothecin / Q000494:pharmacology; D053148:Caspase 3 / Q000378:metabolism*; D053179:Caspase 7 / Q000378:metabolism*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D003110:Colonic Neoplasms / Q000188:drug therapy / Q000201:enzymology / Q000473:pathology; D003606:Dacarbazine / Q000031:analogs & derivatives / Q000494:pharmacology; D004789:Enzyme Activation; D049454:Firefly Luciferin / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D005434:Flow Cytometry; D005909:Glioblastoma / Q000188:drug therapy / Q000201:enzymology / Q000473:pathology; D006801:Humans; D008163:Luminescent Measurements / Q000379:methods*; D008164:Luminescent Proteins / Q000737:chemistry / Q000378:metabolism; D051379:Mice; D008819:Mice, Nude; D009842:Oligopeptides / Q000737:chemistry / Q000378:metabolism; D013194:Staining and Labeling; D000077204:Temozolomide; D023041:Xenograft Model Antitumor Assays
chemical_list: D008164:Luminescent Proteins; D009842:Oligopeptides; C550610:benzyloxycarbonyl-aspartyl-glutamyl-valyl-aspartyl-aminoluciferin; D049454:Firefly Luciferin; D003606:Dacarbazine; D053148:Caspase 3; D053179:Caspase 7; D002166:Camptothecin; D000077204:Temozolomide