DNA binding chelates for nonviral gene delivery imaging

Citation
A. Bogdanov et al., DNA binding chelates for nonviral gene delivery imaging, GENE THER, 8(7), 2001, pp. 515-522
Citations number
41
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Molecular Biology & Genetics
Journal title
GENE THERAPY
ISSN journal
0969-7128 → ACNP
Volume
8
Issue
7
Year of publication
2001
Pages
515 - 522
Database
ISI
SICI code
0969-7128(200104)8:7<515:DBCFNG>2.0.ZU;2-4
Abstract
Noninvasive in vivo monitoring of gene delivery would provide a critically important information regarding the spatial distribution, local concentrati on, kinetics of removal and/or biodegradation of the expression vector. We developed a novel approach to noninvasive gene delivery imaging using heter obifunctional peptide-based chelates (PBC) bearing double-stranded DNA-bind ing groups and a technetium-binding amino acid motif. One of such chelates: Gly-Cys (Acm)-Gly-Cys(Acm)-Gly-Lys(4)-Lys-(N-epsilon-[4-(psoralen-8-yloxy) ]butyrate)-NH2 has been characterized and labeled with reduced Tc-99m perte chnetate (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 Tc-99m-labele d PBC was nonextractable and coeluted with a model pCMV-GFP vector during t he gel-permeation chromatography. Nuclear imaging of 'naked' DNA and DNA co mplexes with lipid-based transfection reagents ('lipoplexes') has been perf ormed after systemic or local administration of Tc-99m-PBC-labeled DNA in m ice. Imaging results were corroborated with the biodistribution using (TC)- T-99m-PBC and P-32-labeled DNA and lipoplexes. A markedly different biodist ribution of Tc-99m PBC-labeled DNA and lipoplexes was observed with the lat ter being rapidly trapped in the liver, spleen and lung. Tc-99m PBC-DNA was used as an imaging tracer during in vivo transfection of B16 melanoma by l ocal injection of 'naked' Tc-99m PBC-DNA and corresponding lipoplexes. As d emonstrated by nuclear imaging, Tc-99m PBC-DNA lipoplexes showed a slower e limination from the site of injection than Tc-99m PBC-DNA alone. This resul t correlated with a higher expression of marker mRNA and green fluorescent protein as determined using RT-PCR and immunohistochemistry, respectively.