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Optical imaging of transferrin targeted PEI/DNA complexes in living subjects

Gene Therapy volume 10, pages 758–764 (2003)Cite this article

Abstract

Noninvasive optical bioluminescence imaging systems are important tools for evaluating gene expression in vivo for study of individual and temporal variation in a living animal. In this report, we demonstrate that expression of the firefly luciferase reporter gene (fl) delivered by transferrin (Tf) targeted polyethylenimine (PEI) complexes with, or without, poly(ethylene glycol) (PEG) modifications can be imaged in living A/J mice bearing N2A tumors using a cooled charged coupled device (CCD) camera. Tf–PEI–PEG, Tf–PEI, and PEI (positive control) complexes were tail-vein injected and mice were imaged at 5, 24, 48, and 72 h after complex injection. After imaging, the organs were analyzed ex vivo for firefly luciferase protein (FL) activity. The Tf and PEG modified formulations show significantly (P<0.05) higher FL activity in vivo and ex vivo at the tumor as compared to other organs, including the lungs (a site of high expression with PEI, the positive control). Furthermore, the in vivo bioluminescent signal correlated well (R2=0.83) with ex vivo FL activity. These data support that noninvasive imaging of fl reporter expression can be used to monitor the specificity of Tf–PEI and Tf–PEI–PEG polyplex targeting of N2A tumors in A/J mice.

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Acknowledgements

We would like to thank Xiaoman Lewis and Khoi Nguyen for their technical help, David Stout for his help with the optical system, and Wolfgang Roedl and Malgorzata Kursa for providing us with formulations from Germany and Austria. This work is supported in part by National Institutes of Health Grants P50 CA86306 (SSG), R0-1 CA82214 (SSG), and SAIRP R24 CA92865 (SSG), Department of Energy Contract DE-FC03-87ER60615 (SSG).

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Authors and Affiliations

  1. The Crump Institute for Molecular Imaging, USA

    I J Hildebrandt, M Iyer & S S Gambhir

  2. Department of Molecular and Medical Pharmacology, USA

    I J Hildebrandt & S S Gambhir

  3. Department of Pharmacy, Ludwig-Maximillian University, Munich, Germany

    E Wagner

  4. Department of Medical Biochemistry, Vienna University Biocenter, Austria

    E Wagner

  5. UCLA-Jonsson Comprehensive Cancer Center, USA

    S S Gambhir

  6. Department of Biomathematics, UCLA School of Medicine, Los Angeles, CA, USA

    S S Gambhir

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  1. I J Hildebrandt
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Hildebrandt, I., Iyer, M., Wagner, E. et al. Optical imaging of transferrin targeted PEI/DNA complexes in living subjects. Gene Ther 10, 758–764 (2003). https://doi.org/10.1038/sj.gt.3301939

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