Abstract
Gene-directed enzyme prodrug therapy (GDEPT) is a promising therapeutic approach for treating cancers of various phenotypes. This strategy is independent of various other chemotherapeutic drugs used for treating cancers where the drugs are mainly designed to target endogenous cellular mechanisms, which are different in various cancer subtypes. In GDEPT an external enzyme, which is different from the cellular proteins, is expressed to convert the injected prodrug in to a toxic metabolite, that normally kill cancer cells express this protein. Theranostic imaging is an approach used to directly monitor the expression of these gene therapy enzymes while evaluating therapeutic effect. We recently developed a dual-GDEPT system where we combined mutant human herpes simplex thymidine kinase (HSV1sr39TK) and E. coli nitroreductase (NTR) enzyme, to improve therapeutic efficiency of cancer gene therapy by simultaneously injecting two prodrugs at a lower dose. In this approach we use two different prodrugs such as ganciclovir (GCV) and CB1954 to target two different cellular mechanisms to kill cancer cells. The developed dual GDEPT system was highly efficacious than that of either of the system used independently. In this chapter, we describe the complete protocol involved for in vitro and in vivo imaging of therapeutic cancer gene therapy evaluation.
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References
Duarte S, Carle G, Faneca H, de Lima MC, Pierrefite-Carle V (2012) Suicide gene therapy in cancer: where do we stand now? Cancer Lett 324:160–170
Sekar TV, Foygel K, Willmann JK, Paulmurugan R (2013) Dual-therapeutic reporter genes fusion for enhanced cancer gene therapy and imaging. Gene Ther 20:529–537
Sekar TV, Foygel K, Ilovich O, Paulmurugan R (2014) Noninvasive theranostic imaging of HSV1-sr39TK-NTR/GCV-CB1954 dual-prodrug therapy in metastatic lung lesions of MDA-MB-231 triple negative breast cancer in mice. Theranostics 4:460–474
Kelkar SS, Reineke TM (2011) Theranostics: combining imaging and therapy. Bioconjug Chem 22:1879–1903
Penheiter AR, Russell SJ, Carlson SK (2012) The sodium iodide symporter (NIS) as an imaging reporter for gene, viral, and cell-based therapies. Curr Gene Ther 12:33–47
Buursma AR, Rutgers V, Hospers GA, Mulder NH, Vaalburg W, de Vries EF (2006) 18F-FEAU as a radiotracer for herpes simplex virus thymidine kinase gene expression: in-vitro comparison with other PET tracers. Nucl Med Commun 27:25–30
Bar-Shir A, Liu G, Greenberg MM, Bulte JW, Gilad AA (2013) Synthesis of a probe for monitoring HSV1-tk reporter gene expression using chemical exchange saturation transfer MRI. Nat Protoc 8:2380–2391
Ponomarev V, Doubrovin M, Serganova I, Vider J, Shavrin A, Beresten T, Ivanova A, Ageyeva L, Tourkova V, Balatoni J, Bornmann W, Blasberg R, Gelovani Tjuvajev J (2004) A novel triple-modality reporter gene for whole-body fluorescent, bioluminescent, and nuclear noninvasive imaging. Eur J Nucl Med Mol Imaging 31:740–751
Bhaumik S, Sekar TV, Depuy J, Klimash J, Paulmurugan R (2012) Noninvasive optical imaging of nitroreductase gene-directed enzyme prodrug therapy system in living animals. Gene Ther 19:295–302
Jin S, Leach JC, Ye K (2009) Nanoparticle-mediated gene delivery. Methods Mol Biol 544:547–557
Howarth JL, Lee YB, Uney JB (2010) Using viral vectors as gene transfer tools (Cell Biology and Toxicology Special Issue: ETCS-UK 1 day meeting on genetic manipulation of cells). Cell Biol Toxicol 26:1–20
Daya S, Berns KI (2008) Gene therapy using adeno-associated virus vectors. Clin Microbiol Rev 21:583–593
Volpers C, Kochanek S (2004) Adenoviral vectors for gene transfer and therapy. J Gene Med 6:S164–S171
Green NK, Seymour LW (2002) Adenoviral vectors: systemic delivery and tumor targeting. Cancer Gene Ther 9:1036–1042
Kasala D, Choi JW, Kim SW, Yun CO (2014) Utilizing adenovirus vectors for gene delivery in cancer. Expert Opin Drug Deliv 11:379–392
Flotte T, Carter B, Conrad C, Guggino W, Reynolds T, Rosenstein B, Taylor G, Walden S, Wetzel R (1996) A phase I study of an adeno-associated virus-CFTR gene vector in adult CF patients with mild lung disease. Human Gene Ther 7:1145–1159
Pan JG, Zhou X, Luo R, Han RF (2012) The adeno-associated virus-mediated HSV-TK/GCV suicide system: a potential strategy for the treatment of bladder carcinoma. Med Oncol 29:1938–1947
Escors D, Breckpot K (2010) Lentiviral vectors in gene therapy: their current status and future potential. Arch Immunol Ther Exp (Warsz) 58:107–119
Devulapally R, Sekar NM, Sekar TV, Foygel K, Massoud TF, Willmann JK, Paulmurugan R (2015) Polymer nanoparticles mediated codelivery of antimiR-10b and antimiR-21 for achieving triple negative breast cancer therapy. ACS Nano 9:2290–2302
Acknowledgement
We thank the Department of Radiology and Canary Center at Stanford for their facilities and support. The funding support by National Institutes of Health (NIH grant R01 CA161091 and R21 CA185805 to R.P) is gratefully acknowledged. We also thank Dr. Sanjiv Sam Gambhir, Chairman, Department of Radiology, Stanford University, for his constant support. We gratefully acknowledge the use of the SCi3 Core Facility, Stanford University.
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Sekar, T., Paulmurugan, R. (2016). Theranostic Imaging of Cancer Gene Therapy. In: Kim, S. (eds) Bioluminescence. Methods in Molecular Biology, vol 1461. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3813-1_20
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DOI: https://doi.org/10.1007/978-1-4939-3813-1_20
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