Enhancement of gene delivery by an analogue of alpha-MSH in a receptor-independent fashion.
In order to transfect melanoma specifically by receptor-mediated endocytosis we prepared dioctadecyl aminoglycylspermine (lipospermine)--DNA complexes with [Nle(4),D-Phe(7)]-alpha-MSH(4--10), a pseudo-peptide analogue of alpha-melanocyte stimulating hormone (alpha-MSH) linked to a thiol-reactive phospholipid. With these complexes we obtained an up to 70-fold increase of transfection with B16-F1 melanoma cells. However when B16-G4F, an alpha-MSH receptor negative melanoma cell line was transfected, an up to 700-fold increased transfection efficiency was observed. The peptide hormone analogue was equally efficient when it was only mixed with lipospermine--DNA complexes without covalent coupling. In addition to melanoma cells we also obtained up to 30-fold increased transfection with BN cells (embryonic liver cells). Our data show that an alpha-MSH analogue increased transfection independently of the MSH receptor expression but reaches efficiencies approaching those obtained with peptides derived from viral fusion proteins. The absence of targeting of constructs containing [Nle(4),D-Phe(7)]-alpha-MSH(4-10) can probably be attributed due to the relatively modest number of MSH receptors at the surface of melanoma. We suggest, however, that the peptide hormone analogue used in this study has membrane-active properties and could be of interest as helper agent to enhance non-viral gene delivery presumably by endosomal-destabilizing properties.
Chluba
J
J
Laboratoire de Chimie Bioorganique, UMR 7514 CNRS-ULP, Faculté de Pharmacie, 74, route du Rhin, 67400 Illkirch, France. johanna.chluba@pharma.u-strasbg.fr
Lima de Souza
D
D
Frisch
B
B
Schuber
F
F
D000345:Affinity Labels; D000818:Animals; D002704:Chlorates; D004247:DNA / Q000737:chemistry; D018014:Gene Transfer Techniques; D051379:Mice; D011979:Receptors, Pituitary Hormone / Q000032:analysis; D013096:Spermine / Q000031:analogs & derivatives* / Q000737:chemistry; D014407:Tumor Cells, Cultured; D000521:alpha-MSH / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology*
D000345:Affinity Labels; D002704:Chlorates; D011979:Receptors, Pituitary Hormone; C063681:dioctadecylamidospermine; D013096:Spermine; D000521:alpha-MSH; C027756:MSH, 4-Nle-7-Phe-alpha-; C029446:MSH receptor; D004247:DNA; C032706:sodium chlorate