Reducible Amino Acid Based Cationic Lipids as Highly Efficient and Serum‐Tolerant Gene Vectors

Zheng, Lei; Yi, Wen-Jing; Su, Rongchuan; Liu, Qiang; Zhao, Zhigang

doi:10.1002/cplu.201500307

Cited by 27 publications

(44 citation statements)

References 41 publications

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“…Dendrimers [ 12 , 13 ], polymers [ 14 , 15 ] and lipids [ 16 , 17 ] are other nanocarriers among non-viral vectors that may incorporate natural amino acids in their structures to reduce cytotoxicity and improve cellular uptake. Lysine, arginine and histidine are some of the most commonly used amino acids because of their positive charge at physiological pH, which enables electrostatic interactions with the anionic nucleic acids and the negatively charged cellular membranes [ 16 , 17 , 18 , 19 , 20 ]. In particular, the histidine group also offers a mechanism for endosomal scape, known as the “proton sponge effect” [ 21 , 22 , 23 ].…”

Section: Introductionmentioning

confidence: 99%

Protein Expression Knockdown in Cancer Cells Induced by a Gemini Cationic Lipid Nanovector with Histidine-Based Polar Heads

et al. 2020

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A histidine-based gemini cationic lipid, which had already demonstrated its efficiency as a plasmid DNA (pDNA) nanocarrier, has been used in this work to transfect a small interfering RNA (siRNA) into cancer cells. In combination with the helper lipid monoolein glycerol (MOG), the cationic lipid was used as an antiGFP-siRNA nanovector in a multidisciplinary study. Initially, a biophysical characterization by zeta potential (ζ) and agarose gel electrophoresis experiments was performed to determine the lipid effective charge and confirm siRNA compaction. The lipoplexes formed were arranged in Lα lamellar lyotropic liquid crystal phases with a cluster-type morphology, as cryo-transmission electron microscopy (cryo-TEM) and small-angle X-ray scattering (SAXS) studies revealed. Additionally, in vitro experiments confirmed the high gene knockdown efficiency of the lipid-based nanovehicle as detected by flow cytometry (FC) and epifluorescence microscopy, even better than that of Lipofectamine2000*, the transfecting reagent commonly used as a positive control. Cytotoxicity assays indicated that the nanovector is non-toxic to cells. Finally, using nano-liquid chromatography tandem mass spectrometry (nanoLC-MS/MS), apolipoprotein A-I and A-II followed by serum albumin were identified as the proteins with higher affinity for the surface of the lipoplexes. This fact could be beyond the remarkable silencing activity of the histidine-based lipid nanocarrier herein presented.

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Section: Introductionmentioning

confidence: 99%

Protein Expression Knockdown in Cancer Cells Induced by a Gemini Cationic Lipid Nanovector with Histidine-Based Polar Heads

et al. 2020

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“…Since the presence of amino acid residues improves the biocompatibility and cationic lipids are able to establish electrostatic interactions with DNA, some studies have been reported on their combination to design and synthesize alternative candidates for plasmid DNA/small interfering RNA nanocarriers [33][34][35][36]. Mono- [37][38][39], di- [38,40,41], tri- [38,41], or multi-peptide [41] cationic lipid systems have been used in in vitro studies for DNA delivery. In mono-histidine cationic lipids [38] and lysine cationic lipid assemblies [34], moderate-to-high transfection efficiencies have been achieved.…”

Section: Introductionmentioning

confidence: 99%

A Non-Viral Plasmid DNA Delivery System Consisting on a Lysine-Derived Cationic Lipid Mixed with a Fusogenic Lipid

et al. 2019

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The insertion of biocompatible amino acid moieties in non-viral gene nanocarriers is an attractive approach that has been recently gaining interest. In this work, a cationic lipid, consisting of a lysine-derived moiety linked to a C12 chain (LYCl) was combined with a common fusogenic helper lipid (DOPE) and evaluated as a potential vehicle to transfect two plasmid DNAs (encoding green fluorescent protein GFP and luciferase) into COS-7 cells. A multidisciplinary approach has been followed: (i) biophysical characterization based on zeta potential, gel electrophoresis, small-angle X-ray scattering (SAXS), and cryo-transmission electronic microscopy (cryo-TEM); (ii) biological studies by fluorescence assisted cell sorting (FACS), luminometry, and cytotoxicity experiments; and (iii) a computational study of the formation of lipid bilayers and their subsequent stabilization with DNA. The results indicate that LYCl/DOPE nanocarriers are capable of compacting the pDNAs and protecting them efficiently against DNase I degradation, by forming Lα lyotropic liquid crystal phases, with an average size of ~200 nm and low polydispersity that facilitate the cellular uptake process. The computational results confirmed that the LYCl/DOPE lipid bilayers are stable and also capable of stabilizing DNA fragments via lipoplex formation, with dimensions consistent with experimental values. The optimum formulations (found at 20% of LYCl content) were able to complete the transfection process efficiently and with high cell viabilities, even improving the outcomes of the positive control Lipo2000*.

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“…Amino acids play important roles in cell life, acting as cell signaling molecules, regulating gene expression, and synthesizing hormones, among others. These features make them attractive as components of amino acid-based gene carriers for gene therapy, or as a part of different kind of molecules, such as of cationic monovalent or multivalent lipids [1,2,3,4,5,6], polypeptides [7,8,9,10,11], polymers [12,13,14,15], polycations [16] and/or dendrimers [11,17,18,19,20]. Generally, gene therapy has focused on viral and non-viral vectors for the treatment of hereditary or acquired illnesses [21,22,23].…”

Section: Introductionmentioning

confidence: 99%

A Gemini Cationic Lipid with Histidine Residues as a Novel Lipid-Based Gene Nanocarrier: A Biophysical and Biochemical Study

et al. 2018

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This work reports the synthesis of a novel gemini cationic lipid that incorporates two histidine-type head groups (C3(C16His)2). Mixed with a helper lipid 1,2-dioleoyl-sn-glycero-3-phosphatidyl ethanol amine (DOPE), it was used to transfect three different types of plasmid DNA: one encoding the green fluorescence protein (pEGFP-C3), one encoding a luciferase (pCMV-Luc), and a therapeutic anti-tumoral agent encoding interleukin-12 (pCMV-IL12). Complementary biophysical experiments (zeta potential, gel electrophoresis, small-angle X-ray scattering (SAXS), and fluorescence anisotropy) and biological studies (FACS, luminometry, and cytotoxicity) of these C3(C16His)2/DOPE-pDNA lipoplexes provided vast insight into their outcomes as gene carriers. They were found to efficiently compact and protect pDNA against DNase I degradation by forming nanoaggregates of 120–290 nm in size, which were further characterized as very fluidic lamellar structures based in a sandwich-type phase, with alternating layers of mixed lipids and an aqueous monolayer where the pDNA and counterions are located. The optimum formulations of these nanoaggregates were able to transfect the pDNAs into COS-7 and HeLa cells with high cell viability, comparable or superior to that of the standard Lipo2000*. The vast amount of information collected from the in vitro studies points to this histidine-based lipid nanocarrier as a potentially interesting candidate for future in vivo studies investigating specific gene therapies.

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Reducible Amino Acid Based Cationic Lipids as Highly Efficient and Serum‐Tolerant Gene Vectors

Abstract: Scheme1.Synthesis of lipids 1a and 1b.Reagents and conditions: a) Cystamine, triethylamine;b)N a -Boc-l-histidine or N a ,N e -di-Boc-l-lysine, HOBt, EDC·HCl, N,N-diisopropylethylamine (DIEA); c) CF 3 COOH, CH 2 Cl 2 .

Cited by 27 publications

References 41 publications

Protein Expression Knockdown in Cancer Cells Induced by a Gemini Cationic Lipid Nanovector with Histidine-Based Polar Heads

Protein Expression Knockdown in Cancer Cells Induced by a Gemini Cationic Lipid Nanovector with Histidine-Based Polar Heads

A Non-Viral Plasmid DNA Delivery System Consisting on a Lysine-Derived Cationic Lipid Mixed with a Fusogenic Lipid

A Gemini Cationic Lipid with Histidine Residues as a Novel Lipid-Based Gene Nanocarrier: A Biophysical and Biochemical Study

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