Aspects of nonviral gene therapy: Correlation of molecular parameters with lipoplex structure and transfection efficacy in pyridinium-based cationic lipids

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• 作者：Paria Parvizi ; Emile Jubeli ; Liji Raju ; Nada Abdul Khalique ; Ahmed Almeer ; Hebatalla Allam ; Maryem Al Manaa ; Helge Larsen ; David Nicholson ; Michael D. Pungente ; Thomas M. Fyles
• 关键词：Cationic lipids ; Pyridinium-based lipids ; Shape parameter ; SAXS ; Lipoplex formulation ; DNA transfer ; Gene therapy.
• 刊名：International Journal of Pharmaceutics
• 出版年：30 January, 2014
• 年：2014
• 卷：461
• 期：1-2
• 页码：145-156
• 全文大小：1999 K

文摘

This study seeks correlations between the molecular structures of cationic and neutral lipids, the lipid phase behavior of the mixed-lipid lipoplexes they form with plasmid DNA, and the transfection efficacy of the lipoplexes. Synthetic cationic pyridinium lipids were co-formulated (1:1) with the cationic lipid 1,2-dimyristoyl-sn-glycero-3-ethylphosphocholine (EPC), and these lipids were co-formulated (3:2) with the neutral lipids 1,2-dioleoyl-sn-glycero-3-phosphatidylethanolamine (DOPE) or cholesterol. All lipoplex formulations exhibited plasmid DNA binding and a level of protection from DNase I degradation. Composition-dependent transfection (beta-galactosidase and GFP) and cytotoxicity was observed in Chinese hamster ovarian-K1 cells. The most active formulations containing the pyridinium lipids were less cytotoxic but of comparable activity to a Lipofectamine 2000鈩?control. Molecular structure parameters and partition coefficients were calculated for all lipids using fragment additive methods. The derived shape parameter values correctly correlated with observed hexagonal lipid phase behavior of lipoplexes as derived from small-angle X-ray scattering experiments. A transfection index applicable to hexagonal phase lipoplexes derived from calculated parameters of the lipid mixture (partition coefficient, shape parameter, lipoplex packing) produced a direct correlation with transfection efficiency.