Fatty acid-spermine conjugates as DNA carriers for nonviral in vivo gene delivery.
The lack of efficient in vivo gene delivery is a well-known shortcoming of nonviral delivery vectors, in particular of chemical vectors. We developed a series of novel nonviral carriers for plasmid-based in vivo gene delivery. This new transport device is based on the assembly of DNA plasmids with synthetic derivatives of naturally occurring molecules-fatty acid-spermine conjugates (or lipospermines). We tested the ability of these fatty acid conjugates to interact with plasmid DNA (pDNA) and found that they formed DNA nanocomplexes, which are protected from DNase I degradation. This protection was shown to directly correlate with the length of the aliphatic component. However, this increase in the length of the hydrocarbon chain resulted in increased toxicity. The cationic lipids used for transfection typically have a C(16) and C(18) hydrocarbon chain. Interestingly, toxicity studies, together with further characterization studies, suggested that the two most suitable candidates for in vivo delivery are those with the shortest hydrocarbon chain, butanoyl- and decanoylspermine. Morphological characterization of DNA nanocomplexes resulting from these lipospermines showed the formation of a homogenous population, with the diameter ranging approximately from 40 to 200 nm. Butanoylspermine was found to be the most promising carrier from this series, resulting in a significantly increased gene expression, in relation to naked plasmid, in both tissues herein targeted (dermis and M. tibialis anterior). Thus, we established a correlation between the in vitro properties of the ensuing DNA nanocarriers and their efficient in vivo gene expression.
Viola
J R
JR
Clinical Research Center, Department of Laboratory Medicine, Karolinska Institutet, Karolinska University Hospital Huddinge, Stockholm, Sweden. Joana.Viola@ki.se
Leijonmarck
H
H
Simonson
O E
OE
Oprea
I I
II
Frithiof
R
R
Purhonen
P
P
Moreno
P M D
PM
Lundin
K E
KE
Strömberg
R
R
Smith
C I E
CI
D000818:Animals; D004247:DNA / Q000737:chemistry; D005227:Fatty Acids / Q000494:pharmacology / Q000633:toxicity; D005231:Fatty Acids, Unsaturated / Q000494:pharmacology / Q000633:toxicity; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000096:biosynthesis / Q000633:toxicity; D056945:Hep G2 Cells; D006801:Humans; D051379:Mice; D010957:Plasmids; D013096:Spermine / Q000031:analogs & derivatives / Q000494:pharmacology / Q000633:toxicity
D005227:Fatty Acids; D005231:Fatty Acids, Unsaturated; D013096:Spermine; D004247:DNA