The role of organ vascularization and lipoplex-serum initial contact in intravenous murine lipofection.
Following intravenous administration of cationic lipid-DNA complexes (lipoplexes) into mice, transfection (lipofection) occurs predominantly in the lungs. This was attributed to high entrapment of lipoplexes in the extended lung vascular tree. To determine whether lipofection in other organs could be enhanced by increasing the degree of vascularization, we used a transgenic mouse model with tissue-specific angiogenesis in liver. Tail vein injection of N-(1-(2,3-dioleoyloxy)propyl)-N,N,N-trimethylammonium chloride (DOTAP)/cholesterol lipoplexes resulted in increased lipoplex entrapment in hypervascularized liver but did not boost luciferase expression, suggesting that lipoplex delivery is not a sufficient condition for efficient organ lipofection. Because the intravenously injected lipoplexes migrated within seconds to lungs, we checked whether the effects of immediate contact with serum correlate with lung lipofection efficiency of different DOTAP-based formulations. Under conditions mimicking the injection environment, the lipoplex-serum interaction was strongly dependent on helper lipid and ionic strength: lipoplexes prepared in 150 mM NaCl or lipoplexes with high (>33 mol%) cholesterol were found to aggregate immediately. This aggregation process was irreversible and was inversely correlated with the percentage of lung cells that took up lipoplexes and with the efficiency of lipofection. No other structural changes in serum were observed for cholesterol-based lipoplexes. Dioleoyl phosphatidylethanolamine-based lipoplexes were found to give low expression, apparently because of an immediate loss of integrity in serum, without lipid-DNA dissociation. Our study suggests that efficient in vivo lipofection is the result of cross-talk between lipoplex composition, interaction with serum, hemodynamics, and target tissue "susceptibility" to transfection.
Simberg
Dmitri
D
Laboratory of Membrane and Liposome Research, Department of Biochemistry, The Hebrew University-Hadassah Medical School, P. O. Box 12272, Jerusalem 91120, Israel.
Weisman
Sarah
S
Talmon
Yeshayahu
Y
Faerman
Alexander
A
Shoshani
Tzipora
T
Barenholz
Yechezkel
Y
D000818:Animals; D001693:Biological Transport, Active; D002784:Cholesterol / Q000008:administration & dosage; D004274:DNA, Recombinant / Q000008:administration & dosage* / Q000097:blood / Q000235:genetics; D016503:Drug Delivery Systems; D005229:Fatty Acids, Monounsaturated / Q000008:administration & dosage* / Q000097:blood; D015870:Gene Expression; D007275:Injections, Intravenous; D008081:Liposomes / Q000008:administration & dosage / Q000097:blood; D008099:Liver / Q000098:blood supply / Q000378:metabolism; D008156:Luciferases / Q000235:genetics; D008168:Lung / Q000098:blood supply / Q000378:metabolism; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008822:Mice, Transgenic; D000644:Quaternary Ammonium Compounds / Q000008:administration & dosage* / Q000097:blood; D014162:Transfection / Q000379:methods*
D004274:DNA, Recombinant; D005229:Fatty Acids, Monounsaturated; D008081:Liposomes; D000644:Quaternary Ammonium Compounds; D002784:Cholesterol; D008156:Luciferases; C070046:1,2-dioleoyloxy-3-(trimethylammonium)propane