Browse > Article
http://dx.doi.org/10.5352/JLS.2021.31.11.1028

Study on the Safety of Firefly Luciferase in Human as a Transient Reporter Gene of Oncolytic Virotherapy  

Hong, Young Mi (Liver Center, Pusan National University Yangsan Hospital, Department of Internal Medicine, School of Medicine, Pusan National University)
Yoon, Woong Hee (Bionoxx Inc.)
Lee, You Ra (Bionoxx Inc.)
Kim, Soo Ji (Bionoxx Inc.)
Ngabire, Daniel (Bionoxx Inc.)
Narayanasamy, Badrinath (Medical Research Center, School of Medicine, Pusan National University)
Ornella, Mefotse Saha Cyrelle (Medical Research Center, School of Medicine, Pusan National University)
Kim, Myunghee (Bionoxx Inc.)
Cho, Euna (Bionoxx Inc.)
Lee, Bora (Bionoxx Inc.)
Hwang, Tae-Ho (Liver Center, Pusan National University Yangsan Hospital, Department of Internal Medicine, School of Medicine, Pusan National University)
Publication Information
Journal of Life Science / v.31, no.11, 2021 , pp. 1028-1036 More about this Journal
Abstract
Firefly luciferase (FLuc) can function as an efficient marker in the gene and viral therapies. Nonetheless, its clinical translation has been unaccomplished with the concerns on its exogenous nature and the similarity with human fatty acyl-CoA synthetase. In this study, we aimed to show safety of FLuc by conducting a set of preclinical experiments and a human use. Initially, FLuc permeability across the plasma membrane was investigated by delivering the FLuc-carrying viral vector, OTS-412, or the FLuc recombinant protein. After in vitro infection of OTS-412 into different cancer cell lines, FLuc activity was detected only in the cell lysates, but not in culture media. In addition, recombinant FLuc protein further showed the impermeability against the plasma membrane. Similar result was also observed in the in vivo experiment. After being injected into the VX2 tumor-bearing rabbit, the FLuc exclusively resided within the tumor tissue without being detected in the blood plasma or other organs. Human cancer cell lines originated from various organs were lysed and treated to the FLuc, and none of the human substrates was reactive against the FLuc. As a final step, FLuc recombinant protein was intravenously injected into a human. The luciferase was degraded with the half-life of 20 to 30 minutes in blood, and was untraceable from 1.5 hr after the injection. In addition, the blood plasma was nonresponsive against the fatty acids. Hematological analysis was also comparable between the pre- and post-injection. Altogether, our study collectively demonstrates the safety of the firefly luciferase.
Keywords
Firefly luciferase; human trial; oncolytic virus; OTS-412; reporter gene;
Citations & Related Records
연도 인용수 순위
  • Reference
1 European Medicines Agency. 2009. ICH considerations-general principles to address virus and vector shedding.
2 Gambhir, S. S., Bauer, E., Black, M. E., Liang, Q., Kokoris, M. S., Barrio, J. R., Iyer, M., Namavari, M., Phelps, M. E. and Herschman, H. R. 2000. A mutant herpes simplex virus type 1 thymidine kinase reporter gene shows improved sensitivity for imaging reporter gene expression with positron emission tomography. Proc. Natl. Acad. Sci. USA. 97, 2785-2790.   DOI
3 Islam, S. M., Lee, B., Jiang, F., Kim, E. K., Ahn, S. C. and Hwang, T. H. 2020. Engineering and characterization of oncolytic vaccinia virus expressing truncated herpes simplex virus thymidine kinase. Cancers 12, 228.   DOI
4 Davis, J. K. 2003. Self-experimentation. Account. Res. 10, 175-187.   DOI
5 Gould, S. J. and Subramani, S. 1988. Firefly luciferase as a tool in molecular and cell biology. Anal. Biochem. 175, 5-13.   DOI
6 Hanley, B. P., Bains, W. and Church, G. 2019. Review of scientific self-experimentation: ethics history, regulation, scenarios, and views among ethics committees and prominent scientists. Rejuvenation Res. 22, 31-42.   DOI
7 Kim, J. E., Kalimuthu, S. and Ahn, B. C. 2015. In Vivo Cell Tracking with Bioluminescence Imaging. Nucl. Med. Mol. Imaging 49, 3-10.   DOI
8 Minn, A. J., Gupta, G. P., Siegel, P. M., Bos, P. D., Shu, W., Giri, D. D., Viale, A., Olshen, A. B., Gerald, W. L. and Massague, J. 2005. Genes that mediate breast cancer metastasis to lung. Nature 436, 518-524.   DOI
9 Park, B. H., Hwang, T., Liu, T. C., Sze, D. Y., Kim, J. S., Kwon, H. C., Oh, S. Y., Han, S. Y., Yoon, J. H., Hong, S. H., Moon, A., Speth, K., Park, C., Ahn, Y. J., Daneshmand, M., Rhee, B. G., Pinedo, H. M., Bell, J. C. and Kirn, D. H. 2008. Use of a targeted oncolytic poxvirus, JX-594, in patients with refractory primary or metastatic liver cancer: a phase I trial. Lancet Oncol. 9, 533-542.   DOI
10 Tietje, C. and Brouder, A. 2010. International conference on harmonisation of technical requirements for registration of pharmaceuticals for human use. Handb. Transl. Econ. Gov. Regimes 1041-1053.
11 Tarantal, A. F. and Lee, C. C. I. 2010. Long-term luciferase expression monitored by bioluminescence imaging after adeno-associated virus-mediated fetal gene delivery in rhesus monkeys (Macaca mulatta). Hum. Gene Ther. 21, 143-148.   DOI
12 Serganova, I. and Blasberg, R. G. 2019. Molecular imaging with reporter genes: has its promise been delivered? J. Nucl. Med. 60, 1665-1681.   DOI
13 Guggino, W. B., Benson, J., Seagrave, J., Yan, Z., Engelhardt, J., Gao, G., Conlon, T. J. and Cebotaru, L. 2017. A preclinical study in rhesus macaques for cystic fibrosis to assess gene transfer and transduction by AAV1 and AAV5 with a dual-luciferase reporter system. Hum. Gene Ther. Clin. Dev. 28, 145-156.   DOI
14 Kim, J. B., Urban, K., Cochran, E., Lee, S., Ang, A., Rice, B., Bata, A., Campbell, K., Coffee, R., Gorodinsky, A., Lu, Z., Zhou, H., Kishimoto, T. K. and Lassota, P. 2010. Non-invasive detection of a small number of bioluminescent cancer cells in vivo. PLoS One 5, e9364.   DOI
15 Leng, L., Wang, Y., He, N., Wang, D., Zhao, Q., Feng, G., Su, G., Su, W., Xu, Y., Han, Z., Kong, D., Cheng, Z., Xiang, R. and Li, Z. 2014. Molecular imaging for assessment of mesenchymal stem cells mediated breast cancer therapy. Biomaterials 35, 5162-5170.   DOI
16 Rabinovich, B. A., Ye, Y., Etto, T., Chen, J. Q., Levitsky, H. I., Overwijk, W. W., Cooper, L. J. N., Gelovani, J. and Hwu, P. 2008. Visualizing fewer than 10 mouse T cells with an enhanced firefly luciferase in immunocompetent mouse models of cancer. Proc. Natl. Acad. Sci. USA. 105, 14342-14346.   DOI
17 Mell, L. K., Brumund, K. T., Daniels, G. A., Advani, S. J., Zakeri, K., Wright, M. E., Onyeama, S. J., Weisman, R. A., Sanghvi, P. R., Martin, P. J. and Szalay, A. A. 2017. Phase I trial of intravenous oncolytic vaccinia virus (GL-ONC1) with cisplatin and radiotherapy in patients with locoregionally advanced head and neck carcinoma. Clin. Cancer Res. 23, 5696-5702.   DOI
18 O'Leary, M. P., Warner, S. G., Kim, S. I., Chaurasiya, S., Lu, J., Choi, A. H., Park, A. K., Woo, Y., Fong, Y. and Chen, N. G. 2018. A novel oncolytic chimeric orthopoxvirus encoding luciferase enables real-time view of colorectal cancer cell infection. Mol. Ther. 9, 13-21.
19 Oba, Y., Ojika, M. and Inouye, S. 2003. Firefly luciferase is a bifunctional enzyme: ATP-dependent monooxygenase and a long chain fatty acyl-CoA synthetase. FEBS Lett. 540, 251-254.   DOI
20 Pozzo, T., Akter, F., Nomura, Y., Louie, A. Y. and Yokobayashi, Y. 2018. Firefly Luciferase Mutant with Enhanced Activity and Thermostability. ACS Omega 3, 2628-2633.   DOI
21 Thorne, N., Inglese, J. and Auld, D. S. 2010. Illuminating insights into firefly luciferase and other bioluminescent reporters used in chemical biology. Chem. Biol. 17, 646-657.   DOI
22 Youn, H. and Chung, J. K. 2013. Reporter gene imaging. AJR. Am. J. Roentgenol. 201, W206-W214.   DOI
23 Chievitz, O. and Hevesy, G. 1935. Radioactive indicators in the study of phosphorous metabolism in rats. Nature 136, 754-755.   DOI
24 Groot-Wassink, T., Aboagye, E. O., Glaser, M., Lemoine, N. R. and Vassaux, G. 2002. Adenovirus biodistribution and noninvasive imaging of gene expression in vivo by positron emission tomography using human sodium/iodide symporter as reporter gene. Hum. Gene Ther. 13, 1723-1735.   DOI
25 Badr, C. E. and Tannous, B. A. 2011. Bioluminescence imaging: progress and applications. Trends Biotechnol. 29, 624-633.   DOI