Characterization of tailor-made copolymers of oligo(ethylene glycol) methyl ether methacrylate and N,N-dimethylaminoethyl methacrylate as nonviral gene transfer agents: influence of macromolecular structure on gene vector particle properties and transfection efficiency.
Oligo(ethylene glycol) methyl ether methacrylates (OEGMA) of various chain lengths (i.e., 9, 23, or 45 EG units) and N,N-dimethylaminoethyl methacrylate (DMAEMA) were copolymerized by atom transfer radical polymerization (ATRP), yielding well-defined P(DMAEMA-co-OEGMA) copolymers with increasing OEGMA molar fractions (F(OEGMA)) but a comparable degree of polymerization (DP approximately 120). Increase of both F(OEGMA) and OEGMA chain lengths correlated inversely with gene vector size, morphology, and zeta potential. P(DMAEMA-co-OEGMA) copolymers prevented gene vector aggregation at high plasmid DNA (pDNA) concentrations in isotonic solution and did not induce cytotoxicity even at high concentrations. Transfection efficiency of the most efficient P(DMAEMA-co-OEGMA) copolymers was found to be >10-fold lower compared with branched polyethylenimine (PEI) 25 kDa. Although OEGMA copolymerization largely reduced gene vector binding with the cell surface, cellular internalization of the bound complexes was less affected. These observations suggest that inefficient endolysosomal escape limits transfection efficiency of P(DMAEMA-co-OEGMA) copolymer gene vectors. Despite this observation, optimized p(DMAEMA-co-OEGMA) gene vectors remained stable under conditions for in vivo application leading to 7-fold greater gene expression in the lungs compared with PEI. Tailor-made P(DMAEMA-co-OEGMA) copolymers are promising nonviral gene transfer agents that fulfill the requirements for successful in vivo gene delivery.
Uzgün
Senta
S
Department of Pediatrics, Ludwig-Maximilians University, 80337 Munich, Germany.
Akdemir
Ozgür
O
Hasenpusch
Günther
G
Maucksch
Christof
C
Golas
Monika M
MM
Sander
Bjoern
B
Stark
Holger
H
Imker
Rabea
R
Lutz
Jean-François
JF
Rudolph
Carsten
C
D000818:Animals; D001980:Bronchi / Q000166:cytology / Q000378:metabolism; D002470:Cell Survival; D004247:DNA / Q000235:genetics / Q000378:metabolism; D004337:Drug Carriers / Q000737:chemistry*; D004587:Electrophoresis, Agar Gel; D004847:Epithelial Cells / Q000378:metabolism; D005260:Female; D005434:Flow Cytometry; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000008:administration & dosage* / Q000494:pharmacology*; D006801:Humans; D008156:Luciferases / Q000378:metabolism; D008689:Methacrylates / Q000737:chemistry*; D020366:Methylmethacrylate / Q000737:chemistry*; D051379:Mice; D008807:Mice, Inbred BALB C; D011094:Polyethyleneimine; D011108:Polymers / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D011650:Pulmonary Alveoli / Q000166:cytology / Q000378:metabolism; D014162:Transfection
D004337:Drug Carriers; D008689:Methacrylates; D011108:Polymers; D020366:Methylmethacrylate; D011094:Polyethyleneimine; D004247:DNA; D008156:Luciferases; C049840:2-(dimethylamino)ethyl methacrylate