In Vitro Cationic Lipid-Mediated Gene Delivery with Fluorinated Glycerophosphoethanolamine Helper Lipids

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• 作者：Jé ; rô ; me Gaucheron ; Caroline Boulanger ; Catherine Santaella ; Nicolas Sbirrazzuoli ; Otmane Boussif ; and Pierre Vierling
• 刊名：Bioconjugate Chemistry
• 出版年：2001
• 出版时间：November 2001
• 年：2001
• 卷：12
• 期：6
• 页码：949 - 963
• 全文大小：624K
• 年卷期：v.12,no.6(November 2001)
• ISSN：1520-4812

文摘

There is a need for the development of nonviral gene transfer systems with improved and originalproperties. "Fluorinated" lipoplexes are such candidates, as supported by the remarkably higher invitro and in vivo transfection potency found for such fluorinated lipoplexes as compared withconventional ones or even with PEI-based polyplexes (Boussif, O., Gaucheron, J., Boulanger, C.,Santaella, C., Kolbe, H. V. J., Vierling, P. (2001) Enhanced in vitro and in vivo cationic lipid-mediatedgene delivery with a fluorinated glycerophosphoethanolamine helper lipid. J. Gene Med. 3, 109-114).Here, we describe the synthesis of fluorinated glycerophosphoethanolamines (F-PEs), close analoguesof dioleoylphosphatidylethanolamine (DOPE), and report on their lipid helper properties vs that ofDOPE, as in vitro gene transfer components of fluorinated lipoplexes based on pcTG90, DOGS(Transfectam), or DOTAP. To evaluate the contribution of the F-PEs to in vitro lipoplex-mediatedgene transfer, we examined the effect of including the F-PEs in lipoplexes formulated with thesecationic lipids (CL) for various CL:DOPE:F-PE molar ratios [1:(1 - x):x with x = 0, 0.5 and 1; 1:(2 -y):y with y = 0, 1, 1.5, and 2], and various N/P ratios (from 10 to 0.8, N = number of CL amines, P =number of DNA phosphates). Irrespective of the F-PE chemical structure, of the colipid F-PE:DOPEcomposition, and of the N/P ratio, comparable transfection levels to those of their respective controlDOPE lipoplexes were most frequently obtained when using one of the F-PEs as colipid of DOGS,pcTG90, or DOTAP in place of part of or of all DOPE. However, a large proportion of DOGS-basedlipoplexes were found to display a higher transfection efficiency when formulated with the F-PEsrather than with DOPE alone while the opposite tendency was evidenced for the DOTAP-basedlipoplexes. The present work indicates that "fluorinated" lipoplexes formulated with fluorinated helperlipids and conventional cationic lipids are very attractive candidates for gene delivery. It confirmsfurther that lipophobicity and restricted miscibility of the lipoplex lipids with the endogenous lipidsdoes not preclude efficient gene transfer and expression. Their transfection potency is ratherattributable to their unique lipophobic and hydrophobic character (resulting from the formulation ofDNA with fluorinated lipids), thus preventing to some extent DNA from interactions with lipophilicand hydrophilic biocompounds, and from degradation.