Activation of the complement system by synthetic DNA complexes: a potential barrier for intravenous gene delivery.
We have examined the complement-activating properties of synthetic cationic molecules and their complexes with DNA. Commonly used gene delivery vehicles include complexes of DNA with polylysine of various chain lengths, transferrin-polylysine, a fifth-generation poly(amidoamine) (PAMAM) dendrimer, poly(ethyleneimine), and several cationic lipids (DOTAP, DC-Chol/DOPE, DOGS/DOPE, and DOTMA/DOPE). These agents activate the complement system to varying extents. Strong complement activation is seen with long-chain polylysines, the dendrimer, poly(ethyleneimine), and DOGS (half-maximal at about 3 microM amine content in the assay used). Compared to these compounds, the other cationic lipids (in liposome formulations) are weak activators of the complement system (half-maximal approximately 50-100 microM positive charge in assay). Complement activation by polylysine is strongly dependent on the chain length. Short-chain oligolysines are comparable to cationic lipids in their activation of complement. Incubation of these compounds with DNA to form complexes reduces complement activation in virtually all cases. The degree of complement activation by DNA complexes is strongly dependent on the ratio of polycation and DNA (expressed as the charge ratio) for polylysine, dendrimer, poly(ethyleneimine), and DOGS. To a lesser degree, charge ratio also influences complement activation by monovalent cationic lipid-DNA complexes. For polylysine-DNA complexes, complement activation can be considerably reduced by modifying the surface of preformed DNA complexes with polyethyleneglycol (half-maximal approximately 20 microM amine content). The data suggests that, by appropriate formulation of DNA complexes, complement activation can be minimized or even avoided. These findings should facilitate the search for DNA complex formulations appropriate for reproducible intravenous gene delivery.
Plank
C
C
University of California, School of Pharmacy, San Francisco 94143-0446, USA.
Mechtler
K
K
Szoka
F C
FC
Wagner
E
E
D000818:Animals; D002412:Cations / Q000737:chemistry / Q000494:pharmacology*; D003167:Complement Activation / Q000187:drug effects*; D004274:DNA, Recombinant / Q000008:administration & dosage / Q000138:chemical synthesis / Q000494:pharmacology*; D005229:Fatty Acids, Monounsaturated / Q000737:chemistry / Q000494:pharmacology; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000008:administration & dosage / Q000737:chemistry / Q000494:pharmacology*; D005998:Glycine / Q000031:analogs & derivatives / Q000737:chemistry / Q000494:pharmacology; D006801:Humans; D007275:Injections, Intravenous; D008081:Liposomes / Q000737:chemistry; D010714:Phosphatidylethanolamines / Q000737:chemistry / Q000494:pharmacology; D010743:Phospholipids / Q000737:chemistry / Q000494:pharmacology*; D011107:Polylysine / Q000737:chemistry / Q000494:pharmacology; D000644:Quaternary Ammonium Compounds / Q000737:chemistry / Q000494:pharmacology; D012756:Sheep / Q000097:blood; D013096:Spermine / Q000031:analogs & derivatives / Q000737:chemistry / Q000494:pharmacology
D002412:Cations; D004274:DNA, Recombinant; D005229:Fatty Acids, Monounsaturated; D008081:Liposomes; D010714:Phosphatidylethanolamines; D010743:Phospholipids; D000644:Quaternary Ammonium Compounds; C054005:N-(1-(2,3-dioleyloxy)propyl)-N,N,N-trimethylammonium; C061134:dioctadecylamidoglycylspermine; D011107:Polylysine; D013096:Spermine; C020888:1,2-dielaidoylphosphatidylethanolamine; C070046:1,2-dioleoyloxy-3-(trimethylammonium)propane; D005998:Glycine