Abstract
This chapter describes the use of whole-body bioluminescent imaging (BLI) for the study of bacterial trafficking in live mice, with an emphasis on the use of bacteria in therapy of cancer. Bacteria present an attractive class of vector for cancer therapy, possessing a natural ability to grow preferentially within tumors following systemic administration. Bacteria engineered to express the lux gene cassette permit BLI detection of the bacteria and tumor sites concurrently. The location and levels of bacteria within tumors over time can be readily examined, visualized in two or three dimensions. The method is applicable to a wide range of bacterial species and tumor xenograft types. This article describes the protocol for analysis of bioluminescent bacteria within subcutaneous tumor-bearing mice. This powerful, and inexpensive, real-time imaging strategy represents an ideal method for the study of bacteria in vivo in the context of cancer research. This protocol outlines the procedure for studying lux-tagged Escherichia coli and Bifidobacterium breve in mice, demonstrating the spatial and temporal readout from 2D and 3D BLI achievable with whole-body in vivo luminescence imaging.
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References
Tangney M, Francis KP (2012) In vivo optical imaging in gene & cell therapy. Curr Gene Ther 12:2–11
Tangney M, Ahmad S, Collins SA, O’Sullivan GC (2010) Gene therapy for prostate cancer. Postgrad Med 122:166–180
Morrissey D, O’Sullivan GC, Tangney M (2010) Tumor targeting with systemically administered bacteria. Curr Gene Ther 10:3–14
Collins SA, Guinn BA, Harrison PT, Scallan MF, O’Sullivan GC, Tangney M (2008) Viral vectors in cancer immunotherapy: which vector for which strategy? Curr Gene Ther 8:66–78
Baban CK, Cronin M, O’Hanlon D, O’Sullivan GC, Tangney M (2010) Bacteria as vectors for gene therapy of cancer. Bioeng Bugs 1:385–394
Yu YA, Zhang Q, Szalay AA (2008) Establishment and characterization of conditions required for tumor colonization by intravenously delivered bacteria. Biotechnol Bioeng 100:567–578
Cronin M, Morrissey D, Rajendran S, El Mashad SM, van Sinderen D, O’Sullivan GC et al (2010) Orally administered bifidobacteria as vehicles for delivery of agents to systemic tumors. Mol Ther 18:1397–1407
van Pijkeren JP, Morrissey D, Monk IR, Cronin M, Rajendran S, O’Sullivan GC et al (2010) A novel Listeria monocytogenes-based DNA delivery system for cancer gene therapy. Hum Gene Ther 21:405–416
Cronin M, Akin AR, Collins SA, Meganck J, Kim JB, Baban CK et al (2012) High resolution in vivo bioluminescent imaging for the study of bacterial tumor targeting. PLoS One 7:e30940
Ahmad S, Casey G, Cronin M, Rajendran S, Sweeney P, Tangney M et al (2011) Induction of effective antitumor response after mucosal bacterial vector mediated DNA vaccination with endogenous prostate cancer specific antigen. J Urol 186:687–693
Riedel CU, Monk IR, Casey PG, Morrissey D, O’Sullivan GC, Tangney M et al (2007) Improved luciferase tagging system for Listeria monocytogenes allows real-time monitoring in vivo and in vitro. Appl Environ Microbiol 73:3091–3094
Baban CK, Cronin M, Akin AR, O’Brien A, Gao X, Tabirca S et al (2012) Bioluminescent bacterial imaging in vivo. J Vis Exp 4:e4318
Cheng CM, Lu YL, Chuang KH, Hung WC, Shiea J, Su YC et al (2008) Tumor-targeting prodrug-activating bacteria for cancer therapy. Cancer Gene Ther 15:393–401
Cronin M, Sleator RD, Hill C, Fitzgerald GF, van Sinderen D (2008) Development of a luciferase-based reporter system to monitor Bifidobacterium breve UCC2003 persistence in mice. BMC Microbiol 8:161
Kuo C, Coquoz O, Troy TL, Xu H, Rice BW (2007) Three-dimensional reconstruction of in vivo bioluminescent sources based on multispectral imaging. J Biomed Opt 12:024007
Acknowledgments
The authors wish to acknowledge Dr. Cormac Gahan and Dr. Susan Joyce for providing lux-tagged E. coli MG1655 and relevant funding support from the European Commission Seventh Framework Programme (PIAP- GA-2013-612219-VIP), Science Foundation Ireland/Enterprise Ireland (12/TIDA/B2437), the Irish Health Research Board (HRA_POR/2010/138), and the Irish Cancer Society (CRF11CRO and PCI12TAN).
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Cronin, M., Akin, A.R., Francis, K.P., Tangney, M. (2016). In Vivo Bioluminescence Imaging of Intratumoral Bacteria. In: Hoffman, R. (eds) Bacterial Therapy of Cancer. Methods in Molecular Biology, vol 1409. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3515-4_7
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DOI: https://doi.org/10.1007/978-1-4939-3515-4_7
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Online ISBN: 978-1-4939-3515-4
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