Polycationic amphiphilic cyclodextrins for gene delivery: synthesis and effect of structural modifications on plasmid DNA complex stability, cytotoxicity, and gene expression.
A molecular-diversity-oriented approach for the preparation of well-defined polycationic amphiphilic cyclodextrins (paCDs) as gene-delivery systems is reported. The synthetic strategy takes advantage of the differential reactivity of primary versus secondary hydroxyl groups on the CD torus to regioselectively decorate each rim with cationic elements and lipophilic tails, respectively. Both the charge density and the hydrophobic-hydrophilic balance can be finely tuned in a highly symmetrical architecture that is reminiscent of both cationic lipids and cationic polymers, the two most prominent types of nonviral gene vectors. The monodisperse nature of paCDs and the modularity of the synthetic scheme are particularly well suited for structure-activity relationship studies. Gel electrophoresis revealed that paCDs self-assemble in the presence of plasmid DNA (pDNA) to provide homogeneous, stable nanoparticles (CDplexes) of 70-150 nm that fully protect pDNA from the environment. The transfection efficiency of the resulting CDplexes has been investigated in vitro on BNL-CL2 and COS-7 cell lines in the absence and presence of serum and found to be intimately dependent on architectural features. Facial amphiphilicity and the presence of a cluster of cationic and hydrogen-bonding centers for cooperative and reversible complexation of the polyanionic DNA chain is crucial to attain high transgene expression levels with very low toxicity profiles. Further enhancement of gene expression, eventually overcoming that of polyplexes from commercial polyethyleneimine (PEI) polymers (22 kDa), is achieved by building up space-oriented dendritic polycationic constructs.
Díaz-Moscoso
Alejandro
A
Instituto de Investigaciones Químicas, CSIC and Universidad de Sevilla, Av. Américo Vespucio 49, Isla de la Cartuja, 41092 Sevilla, Spain.
Le Gourriérec
Löic
L
Gómez-García
Marta
M
Benito
Juan M
JM
Balbuena
Patricia
P
Ortega-Caballero
Fernando
F
Guilloteau
Nicolas
N
Di Giorgio
Christophe
C
Vierling
Pierre
P
Defaye
Jacques
J
Ortiz Mellet
Carmen
C
García Fernández
José M
JM
D000818:Animals; D002460:Cell Line; D003505:Cyclodextrins / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism* / Q000633:toxicity; D004247:DNA / Q000235:genetics* / Q000378:metabolism*; D015870:Gene Expression; D010957:Plasmids / Q000235:genetics* / Q000378:metabolism*; D011108:Polymers / Q000737:chemistry; D014162:Transfection
D003505:Cyclodextrins; D011108:Polymers; D004247:DNA