Photochemical Internalization of Transgenes Controlled by the Heat-shock Protein 70 Promoter

Photochemical internalization (PCI) is a targeting technique that facilitates endosomal escape of macromolecules, such as transgenes, in response to photochemical treatment with endosome/lysosome-localized photosensitizers, such as disulfonated meso-tetraphenylporphine (TPPS2a). In gene therapy this...

Full description

Saved in:
Bibliographic Details
Published in:Photochemistry and photobiology 2006-05, Vol.82 (3), p.809-816
Main Authors: Prasmickaite, Lina, Hellum, Marit, Kaalhus, Olav, Høgset, Anders, Wagner, Ernst, Berg, Kristian
Format: Article
Language:English
Subjects:
Cell Line, Tumor
Cytomegalovirus
Endocytosis
Gene therapy
Genes
Genetic Therapy - methods
HSP70 Heat-Shock Proteins - genetics
Humans
Photochemistry
Plasmids - administration & dosage
Plasmids - pharmacokinetics
Porphyrins - pharmacology
Promoter Regions, Genetic
Proteins
Transfection - methods
Transgenes
Up-Regulation - drug effects
Up-Regulation - genetics
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Photochemical internalization (PCI) is a targeting technique that facilitates endosomal escape of macromolecules, such as transgenes, in response to photochemical treatment with endosome/lysosome-localized photosensitizers, such as disulfonated meso-tetraphenylporphine (TPPS2a). In gene therapy this leads to enhanced transgene expression. Moreover, photochemical treatment generally activates transcription of stress-response genes, such as heat-shock proteins (HSPs), via stimulation of corresponding promoters. Therefore, we used HSP70 (HSPp; a promoter from the HSP family gene) and investigated whether the PCI stimulus could also activate HSPp and thereby stimulate transcription (expression) of the HSPp-controlled transgene internalized via PCI. Using human colorectal carcinoma and hepatoma cell lines in vitro, we showed that TPPS2a-based photochemical treatment enhances expression of cellular HSP70, which correlated with a photochemically enhanced expression (approximately 2-fold, at PCI-optimal doses) of the HSPp-controlled transgene integrated in the genome. Furthermore, PCI enhanced expression of the HSPp-controlled episomal transgene delivered as a plasmid. However, in plasmid-based transfection, PCI-mediated enhancement with HSPp did not exceed the enhancement achieved with the constitutive active CMV promoter. In conclusion, we demonstrated that the PCI-relevant treatment initiates HSP70 response and that the HSP70 promoter can be used in combination with PCI, leading to PCI-enhanced expression of the HSPp-controlled transgene.
ISSN:0031-8655
1751-1097
DOI:10.1562/2005-11-07-RA-731