Imaging Proteolytic Activities in Mouse Models of Cancer

Overview

Journal Methods Mol Biol

Specialty Molecular Biology

Date 2018 Jan 11

PMID 29318559

Authors

Anupama Pal

Alnawaz Rehemtulla

Affiliations

Soon will be listed here.

Abstract

Proteases are "protein-cleaving" enzymes, which, in addition to their non-specific degrading function, also catalyze the highly specific and regulated process of proteolytic processing, thus regulating multiple biological functions. Alterations in proteolytic activity occur during pathological conditions such as cancer. One of the major deregulated classes of proteases in cancer is caspases, the proteolytic initiators and mediators of the apoptotic machinery. The ability to image apoptosis noninvasively in living cells and animal models of cancer can not only provide new insight into the biological basis of the disease but can also be used as a quantitative tool to screen and evaluate novel therapeutic strategies. Optical molecular imaging such as bioluminescence-based genetically engineered biosensors has been developed in our laboratory and exploited to study protease activity in animal models with a high signal to noise. Using the circularly permuted form of firefly luciferase, we have developed a reporter for Caspase 3/7, referred to as Caspase 3/7 GloSensor. Here, we discuss the use of the Caspase 3/7 GloSensor for imaging apoptotic activity in mouse xenografts and genetically engineered mouse models of cancer and present the potential of this powerful platform technology to image the proteolytic activity of numerous other proteases.

References

Sternlicht M, Werb Z . How matrix metalloproteinases regulate cell behavior. Annu Rev Cell Dev Biol. 2001; 17:463-516. PMC: 2792593. DOI: 10.1146/annurev.cellbio.17.1.463. View

Lopez-Otin C, Overall C . Protease degradomics: a new challenge for proteomics. Nat Rev Mol Cell Biol. 2002; 3(7):509-19. DOI: 10.1038/nrm858. View

Marino G, Uria J, Puente X, Quesada V, Bordallo J, Lopez-Otin C . Human autophagins, a family of cysteine proteinases potentially implicated in cell degradation by autophagy. J Biol Chem. 2002; 278(6):3671-8. DOI: 10.1074/jbc.M208247200. View

Zhivotovsky B . Caspases: the enzymes of death. Essays Biochem. 2003; 39:25-40. DOI: 10.1042/bse0390025. View

Choy G, Choyke P, Libutti S . Current advances in molecular imaging: noninvasive in vivo bioluminescent and fluorescent optical imaging in cancer research. Mol Imaging. 2004; 2(4):303-12. DOI: 10.1162/15353500200303142. View

Luker K, Smith M, Luker G, Gammon S, Piwnica-Worms H, Piwnica-Worms D . Kinetics of regulated protein-protein interactions revealed with firefly luciferase complementation imaging in cells and living animals. Proc Natl Acad Sci U S A. 2004; 101(33):12288-93. PMC: 514471. DOI: 10.1073/pnas.0404041101. View

Sauer R, Bolon D, Burton B, Burton R, Flynn J, Grant R . Sculpting the proteome with AAA(+) proteases and disassembly machines. Cell. 2004; 119(1):9-18. PMC: 2717008. DOI: 10.1016/j.cell.2004.09.020. View

Ehrmann M, Clausen T . Proteolysis as a regulatory mechanism. Annu Rev Genet. 2004; 38:709-24. DOI: 10.1146/annurev.genet.38.072902.093416. View

Hollenberg M . Physiology and pathophysiology of proteinase-activated receptors (PARs): proteinases as hormone-like signal messengers: PARs and more. J Pharmacol Sci. 2005; 97(1):8-13. DOI: 10.1254/jphs.fmj04005x2. View

10.

Sato A, Klaunberg B, Tolwani R . In vivo bioluminescence imaging. Comp Med. 2005; 54(6):631-4. View

11.

Ciechanover A . Proteolysis: from the lysosome to ubiquitin and the proteasome. Nat Rev Mol Cell Biol. 2005; 6(1):79-87. DOI: 10.1038/nrm1552. View

12.

Vermeulen K, Bockstaele D, Berneman Z . Apoptosis: mechanisms and relevance in cancer. Ann Hematol. 2005; 84(10):627-39. DOI: 10.1007/s00277-005-1065-x. View

13.

Siegel R . Caspases at the crossroads of immune-cell life and death. Nat Rev Immunol. 2006; 6(4):308-17. DOI: 10.1038/nri1809. View

14.

Page-McCaw A, Ewald A, Werb Z . Matrix metalloproteinases and the regulation of tissue remodelling. Nat Rev Mol Cell Biol. 2007; 8(3):221-33. PMC: 2760082. DOI: 10.1038/nrm2125. View

15.

Zhang L, Lee K, Bhojani M, Khan A, Shilman A, Holland E . Molecular imaging of Akt kinase activity. Nat Med. 2007; 13(9):1114-9. DOI: 10.1038/nm1608. View

16.

Fan F, Binkowski B, Butler B, Stecha P, Lewis M, Wood K . Novel genetically encoded biosensors using firefly luciferase. ACS Chem Biol. 2008; 3(6):346-51. DOI: 10.1021/cb8000414. View

17.

Lopez-Otin C, Bond J . Proteases: multifunctional enzymes in life and disease. J Biol Chem. 2008; 283(45):30433-7. PMC: 2576539. DOI: 10.1074/jbc.R800035200. View

18.

Edgington L, Berger A, Blum G, Albrow V, Paulick M, Lineberry N . Noninvasive optical imaging of apoptosis by caspase-targeted activity-based probes. Nat Med. 2009; 15(8):967-73. PMC: 3196344. DOI: 10.1038/nm.1938. View

19.

Wigdal S, Anderson J, Vidugiris G, Shultz J, Wood K, Fan F . A novel bioluminescent protease assay using engineered firefly luciferase. Curr Chem Genomics. 2010; 2:16-28. PMC: 2803436. DOI: 10.2174/1875397300802010016. View

20.

Khanna D, Hamilton C, Bhojani M, Lee K, Dlugosz A, Ross B . A transgenic mouse for imaging caspase-dependent apoptosis within the skin. J Invest Dermatol. 2010; 130(7):1797-806. PMC: 3097416. DOI: 10.1038/jid.2010.55. View