Biodistribution study of phosphonolipids: a class of non-viral vectors efficient in mice lung-directed gene transfer.
BACKGROUND
A multitude of cationic lipids have been synthesized since they were first proposed for use in gene therapy. Cationic lipids are able to efficiently transfect cells both in vitro and in vivo. Whereas most research groups have focused their investigations on the toxicity of these molecules, and on the location of expression of the DNA transferred by these vectors, little has been done to determine their biodistribution and elimination pathways. Our group has developed a family of cationic lipids termed phosphonolipids. Following a large in vitro screening experiment, we have selected several molecules for in vivo testing, with some of these phosphonolipids forming lipoplexes efficient in transfecting mouse lungs. It was thus of interest to study their fate after intravenous injection.


METHODS
The respective biodistributions of both the GLB43 phosphonolipid and plasmid DNA were investigated and compared with DNA expression sites. Using the optimal conditions determined for phosphonolipids, we followed the gene transfer agent and plasmid DNA distributions versus time by radiolabeling them with (14)C and (32)P, respectively. Otherwise, we performed imaging by radiolabeling plasmid DNA with (99m)Tc.


RESULTS
The lipoplexes appear to be directly located in the lung after administration. Secondly, the plasmid is released mainly into the lungs and the phosphonolipid vector is rapidly degraded. The hydrophilic moiety of the phosphonolipid is eliminated in the urine, as is the free plasmid.


CONCLUSIONS
This study reveals that there are slight differences in the observed results depending on the technique used to label the DNA; secondly, results show that the residence time of phosphonolipids in the mouse body is related to the DNA binding time.
Delépine
P
P
Inserm EMI-U 01-15: génétique moléculaire et épidémiologie génétique, 46 rue Félix Le Dantec, BP 454, 29275 Brest, France. pascal.delepine@univ-brest.fr
Guillaume
C
C
Montier
T
T
Clément
J C
JC
Yaouanc
J J
JJ
Des Abbayes
H
H
Berthou
F
F
Le Pape
A
A
Férec
C
C
D000818:Animals; D001682:Biological Availability; D005260:Female; D015870:Gene Expression; D018390:Gene Targeting; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors; D007275:Injections, Intravenous; D008081:Liposomes; D008156:Luciferases / Q000378:metabolism; D008168:Lung / Q000378:metabolism*; D051379:Mice; D010743:Phospholipids; D010957:Plasmids
D008081:Liposomes; D010743:Phospholipids; C028299:phosphonolipids; D008156:Luciferases