Liposome transduction into cells enhanced by haptotactic peptides (Haptides) homologous to fibrinogen C-termini.
Haptides are 19-21mer cell-binding peptides equivalent to sequences on the C-termini of fibrinogen beta chain (Cbeta), gamma chain (preCgamma) and the extended alphaE chain of fibrinogen (CalphaE). In solution, Haptides accumulated in cells by non-saturable kinetics [Exp. Cell Res. 287 (2003) 116]. This study describes Haptide interactions with liposomes and Haptide-mediated liposome uptake by cells. Haptides became incorporated into negatively charged liposomes, changing their zeta potential. Atomic force microscopy and particle sizing by light scattering showed that the liposomes dissolved Haptide nanoparticles and absorbed them from solution. Pre-mixing fluorescent rhodamine-containing liposomes or "stealth" doxorubicin (DOX)-containing liposomes (Doxil) with Cbeta, preCgamma or to a lesser degree CalphaE, significantly enhanced their uptake by fibroblasts and endothelial cells. Confocal microscopy showed Haptide-induced liposome uptake saturated above approximately 40 microM Haptide. Cytotoxicity tests with lower concentrations of Doxil liposomes indicated that premixing with approximately 40 microM Cbeta or preCgamma increased their toxicity by one order of magnitude. It was evident that the liposomes complexed with an amphiphilic Haptide are transduced through cell membranes, probably by a non-receptor-mediated process. These results suggest that Cbeta or pre-Cgamma could be employed to augment the cellular uptake of drugs in liposomal formulations.
Gorodetsky
Raphael
R
Laboratory of Radiobiology and Biotechnology, Sharett Institute of Oncology, Hadassah University Hospital, P.O.B. 12000, Jerusalem 91120, Israel. Rafi@Hadassah.org.il
Levdansky
Lila
L
Vexler
Akiva
A
Shimeliovich
Irina
I
Kassis
Ibrahim
I
Ben-Moshe
Matti
M
Magdassi
Shlomo
S
Marx
Gerard
G
D000818:Animals; D001011:Aorta / Q000166:cytology; D002417:Cattle; D002478:Cells, Cultured; D002626:Chemistry, Pharmaceutical; D004317:Doxorubicin / Q000737:chemistry; D004337:Drug Carriers; D005340:Fibrinogen / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism*; D005347:Fibroblasts / Q000166:cytology; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D008081:Liposomes / Q000008:administration & dosage / Q000378:metabolism*; D018613:Microscopy, Confocal / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism*; D012235:Rhodamines / Q000737:chemistry
D004337:Drug Carriers; D005456:Fluorescent Dyes; D008081:Liposomes; D010446:Peptide Fragments; D012235:Rhodamines; D004317:Doxorubicin; D005340:Fibrinogen; D016650:Fluorescein-5-isothiocyanate