Evaluation of apoptosis imaging biomarkers in a genetic model of cell death

Abstract

Purpose We have previously developed the caspase-based radiotracer, 18 F-ICMT-11, for PET imaging to monitor treatment response. We further validated 18 F-ICMT-11 specificity in a murine melanoma death-switch tumour model with conditional activation of caspase-3 induced by doxycycline. Methods Caspase-3/7 activity and cellular uptake of 18 F-ICMT-11, 18 F-ML-10 and 18 F-FDG were assessed in B16ova and B16ovaRevC3 cells after death-switch induction. Death-switch induction was confirmed in vivo in xenograft tumours, and 18 F-ICMT-11 and 18 F-ML-10 biodistribution was assessed by ex vivo gamma counting of select tissues. PET imaging was performed with 18 F-ICMT-11, 18 F-ML-10 and 18 F-FDG. Caspase-3 activation was confirmed by immunohistochemistry. Results Significantly increased caspase-3/7 activity was observed only in B16ovaRevC3 cells after death-switch induction, accompanied by significantly increased 18 F-ICMT-11 ( p < 0.001) and 18 F-ML-10 ( p < 0.05) and decreased 18 F-FDG ( p < 0.001) uptake compared with controls. B16ova and B16ovaRevC3 tumours had similar growth in vivo; however, B16ovaRevC3 growth was significantly reduced with death-switch induction ( p < 0.01). Biodistribution studies showed significantly increased 18 F-ICMT-11 tumour uptake following death-switch induction ( p < 0.01), but not for 18 F-ML-10. Tumour uptake of 18 F-ICMT-11 was higher than that of 18 F-ML-10 after death-switch induction. PET imaging studies showed that 18 F-ICMT-11 can be used to detect apoptosis after death-switch induction, which was accompanied by significantly increased expression of cleaved caspase-3. 18 F-FDG signal decreased in tumours after death-switch induction. Conclusions We demonstrate that 18 F-ICMT-11 can be used to detect caspase-3 activation in a death-switch tumour model, independent of the confounding effects of cancer therapeutics, thus confirming its specificity and supporting the development of this radiotracer for clinical use to monitor tumour apoptosis and therapy response. Electronic supplementary material The online version of this article (10.1186/s13550-019-0487-8) contains supplementary material, which is available to authorized users.

Glutamate and microglia activation as a driver of dendritic apoptosis: a core pathophysiological mechanism to understand schizophrenia

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An Insight into the Mechanism of Holamine- and Funtumine-Induced Cell Death in Cancer Cells

“…The intrinsic and extrinsic apoptotic pathways converge at the point where the executioner caspase, caspase-3 is activated [34]. Therefore, caspase-3 is an important molecular biomarker for the assessment of apoptosis [35]. In this study, caspase-3/7 activity following 12-and 24-h exposure to the compounds was considerably higher in HeLa cells at higher concentrations ( Figure 6).…”

Development of [18F]ICMT-11 for Imaging Caspase-3/7 Activity during Therapy-Induced Apoptosis

“…Induction of this death-switch by doxycycline results in rapid and synchronous apoptosis both in vitro and in vivo in syngeneic murine melanoma lines. The uptakes of [ 18 F]ICMT-11 and the apoptosis radiotracer [ 18 F]ML-10 were evaluated in B16ova and B16ovaRevC3 cells with and without doxycycline [38]. Although the mechanism of action for [ 18 F]ML-10 remains unclear, it is postulated to function through attraction of the anionic head group to positively charged phosphatidylserine residues on the surface of apoptotic cells followed by "anchoring" of the lipophilic tail in the permeable membrane of the dying cell.…”