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Secreted Luciferase for In Vivo Evaluation of Systemic Protein Delivery in Mice

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Abstract

A naturally secreted Gaussia luciferase (Gluc) has been utilized as a reporter for bioluminescence imaging (BLI) evaluation. However, the potential application of Gluc for in vivo monitoring of systemic protein delivery, as well as its natural biodistribution, has not been studied. To examine Gluc secretion and uptake profile, we injected Gluc-encoding plasmids into mice by hydrodynamic tail-vein injection. Whole-body BLI showed that imaging quantification obtained at pawpad was directly correlated to blood Gluc activities. When gene expression was restricted to the liver by the use of a hepatic promoter, in vivo Gluc biodistribution analysis revealed the kidney/bladder, stomach/intestine, and lung as the major uptake organs. Three-dimensional BLI identified liver/stomach and lung as the main internal luminescent sources, demonstrating the feasibility of detecting major uptake organs in live animals by 3D BLI with high-background signals in circulation. Notably, Gluc levels in capillary-depleted brain samples from Gluc-injected mice were comparable to controls, suggesting that Gluc may not cross the blood–brain barrier. Gluc uptake kinetics and intracellular half-life were assessed in various types of cell lines, implicating the involvement of non-specific pinocytosis. These results suggest that Gluc-based system may provide a useful tool for in vivo evaluation of protein/agent biodistribution following systemic delivery.

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Acknowledgments

We would like to thank Amber Edwards and Meghan Bromwell for technical assistance and Comprehensive Mouse and Cancer Core, as well as Veterinary Services Division, for their help with animal manipulation. This work was supported by National Institutes of Health grant NS 064330 (to D.P.).

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Authors and Affiliations

  1. Molecular and Cell Therapy Program, Division of Experimental Hematology & Cancer Biology, Cancer and Blood Diseases Institute, Cincinnati Children’s Hospital Medical Center, Cincinnati, OH, USA

    Salim S. El-Amouri & Dao Pan

  2. Department of Pediatrics, School of Medicine, University of Cincinnati, Cincinnati, OH, USA

    Phuong Cao & Dao Pan

  3. Division of Allergy and Infectious Diseases, University of Washington, Seattle, WA, USA

    Carol Miao

Authors
  1. Salim S. El-Amouri
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  2. Phuong Cao
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  3. Carol Miao
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  4. Dao Pan
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Correspondence to Dao Pan.

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El-Amouri, S.S., Cao, P., Miao, C. et al. Secreted Luciferase for In Vivo Evaluation of Systemic Protein Delivery in Mice. Mol Biotechnol 53, 63–73 (2013). https://doi.org/10.1007/s12033-012-9519-6

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  • DOI: https://doi.org/10.1007/s12033-012-9519-6

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