Lipid-mediated protein delivery of suicide nucleoside kinases.
Nucleoside kinases from several species are investigated as suicide genes for treatment of malignant tumors by combined gene/chemotherapy. In the present study, we have investigated a novel strategy where nucleoside kinase proteins are directly delivered to cells without delivery of genetic material. We used a mix of a trifluoroacetylated lipopolyamine and dioleoyl phosphatidylethanolamine (BioPorter) to form protein-lipid complexes containing either recombinant herpes simplex virus type-1 thymidine kinase or Drosophila melanogaster multisubstrate deoxyribonucleoside kinase. We showed that the nucleoside kinase containing protein-lipid complexes was imported into human osteosarcoma and Chinese hamster ovary cell lines by endocytosis and that the enzymes were delivered to the cytosol and nucleus. The nucleoside kinases imported into the cell lines retained enzymatic activity, and the cells treated with the enzyme-lipid complexes showed increased sensitivity to nucleoside analogues, such as ganciclovir, (E)-5-(2-bromovinyl)-2'-deoxyuridine, and 1-beta-D-arabinofuranosylthymine. Our results show that direct delivery of suicide gene proteins to cells may be an alternative approach to conventional suicide gene therapy strategies.
Division of Clinical Virology F68, Karolinska Institute, Huddinge University Hospital, S-14186 Stockholm, Sweden.
Xinyu
X
Zheng
Mathias
M
Lundberg
Anna
A
Karlsson
Magnus
M
Johansson
D000818:Animals; D001087:Arabinonucleosides / Q000493:pharmacokinetics / Q000494:pharmacology; D001859:Bone Neoplasms / Q000188:drug therapy* / Q000201:enzymology* / Q000378:metabolism / Q000473:pathology; D001973:Bromodeoxyuridine / Q000031:analogs & derivatives* / Q000493:pharmacokinetics / Q000494:pharmacology; D016466:CHO Cells; D045744:Cell Line, Tumor; D006224:Cricetinae; D004331:Drosophila melanogaster / Q000201:enzymology; D004705:Endocytosis; D015774:Ganciclovir / Q000493:pharmacokinetics / Q000494:pharmacology; D018259:Herpesvirus 1, Human / Q000201:enzymology / Q000235:genetics; D006801:Humans; D008081:Liposomes / Q000008:administration & dosage; D012516:Osteosarcoma / Q000188:drug therapy* / Q000201:enzymology* / Q000378:metabolism / Q000473:pathology; D010714:Phosphatidylethanolamines / Q000008:administration & dosage; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000008:administration & dosage* / Q000378:metabolism; D011994:Recombinant Proteins / Q000008:administration & dosage; D013936:Thymidine / Q000031:analogs & derivatives* / Q000493:pharmacokinetics / Q000494:pharmacology; D013937:Thymidine Kinase / Q000008:administration & dosage* / Q000378:metabolism
D001087:Arabinonucleosides; D008081:Liposomes; D010714:Phosphatidylethanolamines; D011994:Recombinant Proteins; C094877:dioleoyl phosphatidylethanolamine; C020235:brivudine; D017853:Phosphotransferases (Alcohol Group Acceptor); C020018:deoxyribonucleoside kinases; D013937:Thymidine Kinase; D001973:Bromodeoxyuridine; C010032:thymine arabinoside; D015774:Ganciclovir; D013936:Thymidine