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DNA binding chelates for nonviral gene delivery imaging

Gene Therapy volume 8, pages 515–522 (2001)Cite this article

Abstract

Noninvasive in vivo monitoring of gene delivery would provide a critically important information regarding the spatial distribution, local concentration, kinetics of removal and/or biodegradation of the expression vector. We developed a novel approach to noninvasive gene delivery imaging using heterobifunctional peptide-based chelates (PBC) bearing double-stranded DNA-binding groups and a technetium-binding amino acid motif. One of such chelates: Gly-Cys(Acm)-Gly-Cys(Acm)-Gly-Lys4-Lys-(N-ε-[4-(psoralen-8-yloxy)]butyrate)-NH2 has been characterized and labeled with reduced 99mtc pertechnetate (oxotechnetate). the psoralen moiety (a dna binding group of pbc) allowed linking to double-stranded dna upon short-term irradiation with the near uv range light (>320 nm). Approximately 30–40% of added 99mTc-labeled PBC was nonextractable and co-eluted with a model pCMV-GFP vector during the gel-permeation chromatography. Nuclear imaging of ‘naked’ DNA and DNA complexes with lipid-based transfection reagents (‘lipoplexes’) has been performed after systemic or local administration of 99mTc-PBC-labeled DNA in mice. Imaging results were corroborated with the biodistribution using 99mTc-PBC and 32P-labeled DNA and lipoplexes. A markedly different biodistribution of 99mTc PBC-labeled DNA and lipoplexes was observed with the latter being rapidly trapped in the liver, spleen and lung. 99mTc PBC-DNA was used as an imaging tracer during in vivo transfection of B16 melanoma by local injection of ‘naked’ 99mTc PBC-DNA and corresponding lipoplexes. As demonstrated by nuclear imaging, 99mTc PBC-DNA lipoplexes showed a slower elimination from the site of injection than 99mTc PBC-DNA alone. This result correlated with a higher expression of marker mRNA and green fluorescent protein as determined using RT-PCR and immunohistochemistry, respectively.

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Acknowledgements

Authors are grateful to Drs Maria Simonova and James Basilion for advice on 32P-labeling. Acknowledged is also the participation of Dr Lee Kayne in the early feasibility experiments. The work was supported in part by NIH: 5RO1 NS35258–03 (RW); 5RO1 CA74424–01 (AB) and 1R21 DK55713–01 (CT).

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  1. S Bredow

    Present address: Lovelace Respiratory Research Institute, POB 5890, Albuquerque, 87185, NM, USA

Authors and Affiliations

  1. Center for Molecular Imaging Research, Massachusetts General Hospital, Charlestown, MA, USA

    A Bogdanov Jr, C-H Tung, S Bredow & R Weissleder

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Bogdanov, A., Tung, CH., Bredow, S. et al. DNA binding chelates for nonviral gene delivery imaging. Gene Ther 8, 515–522 (2001). https://doi.org/10.1038/sj.gt.3301410

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