Multimodal In Vivo Imaging and Blood Monitoring of Intrinsic and Extrinsic Apoptosis
| Autor: | Bakhos A. Tannous, Mariam Kerami, Lisa Pike, Johanna M. Niers, Grant K. Lewandrowski |
|---|---|
| Přispěvatelé: | Neurosurgery, CCA - Disease profiling |
| Rok vydání: | 2011 |
| Předmět: |
Diagnostic Imaging
Blotting Western Green Fluorescent Proteins Golgi Apparatus Mice Nude Apoptosis Caspase 3 Biosensing Techniques Biology Endoplasmic Reticulum Green fluorescent protein Mice 03 medical and health sciences symbols.namesake 0302 clinical medicine In vivo Cell Line Tumor Drug Discovery In Situ Nick-End Labeling Genetics Animals Humans Bioluminescence imaging Molecular Biology Secretory pathway 030304 developmental biology Pharmacology 0303 health sciences Endoplasmic reticulum Glioma Golgi apparatus 3. Good health Cell biology Biochemistry 030220 oncology & carcinogenesis symbols Molecular Medicine Original Article Ex vivo |
| Zdroj: | Molecular Therapy, 19(6), 1090-1096. Nature Publishing Group Niers, J M, Kerami, M, Pike, L, Lewandrowski, G & Tannous, B A 2011, ' Multimodal In Vivo Imaging and Blood Monitoring of Intrinsic and Extrinsic Apoptosis ', Molecular Therapy, vol. 19, no. 6, pp. 1090-1096 . https://doi.org/10.1038/mt.2011.17 |
| ISSN: | 1525-0016 |
| DOI: | 10.1038/mt.2011.17 |
| Popis: | Noninvasive detection and in vivo imaging of apoptosis plays a critical role in the development of therapeutics in many different fields including cancer. We have developed an apoptosis biosensor by fusing green fluorescent protein (GFP) to the N-terminus of the naturally secreted Gaussia luciferase separated by a caspase-3 cleavage peptide consisting of aspartic acid (D), glutamic acid (E), valine (V), and aspartic acid (D) or DEVD. We showed that this fusion is retained in the cytoplasm of cells in an inactive form. Upon apoptosis, the DEVD peptide is cleaved in response to caspase-3 activation, freeing ssGluc, which can now enter the secretory pathway where it is folded properly and is released from the cells and can be detected in the conditioned medium in culture or in blood of live animals ex vivo over time. Because Gluc is secreted from cells via conventional pathway through the endoplasmic reticulum (ER), Golgi and vesicles, we showed that the presence of Gluc in these compartments in response to apoptosis can be visualized in vivo using bioluminescence imaging. This reporter provides a valuable tool for imaging and real-time monitoring of apoptosis and is compatible with high-throughput functional screening application in cultured cells and animal models. |
| Databáze: | OpenAIRE |
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