@article{osti_1046374, title = {Cationic Lipid-Nucleic Acid Complexes for Gene Delivery And Silencing: Pathways And Mechanisms for Plasmid Dna And Sirna}, author = {Ewert, K K and Zidovska, A and Ahmad, A and Bouxsein, N F and Evans, H M and McAllister, C S and Samuel, C E and Safinya, C R and /SLAC}, abstractNote = {Motivated by the promises of gene therapy, there is great interest in . The cationic/ionizable lipids are included to allow the complexing of the negatively charged mRNA molecules and enable the exit of the mRNA from the endosome to the cytosol for translation 3. Here, for the first time, we explored and compared optimal condition (s) of two commercial transfection reagents, Attractene and X-tremeGENE HP, for the transfection of human gastric adenocarcinoma AGS cells. low toxicity, ease of production, and the potential of transferring large pieces of DNA into cells. Conventional liposomal formulations reduced the toxicity of compounds in vivo, through modifying pharmacokinetics and biodistribution to enhance drug delivery to diseased tissue in comparison to free drug. 39 In 1987, Felgner et al. Chronic toxicity means toxicity involving a stimulus that lingers or continues for a relatively long period relative to the life span of an organism. "cellular toxicity of cationic lipids has been linked to increased production of reactive oxygen species."xiv(reactive oxygen species, also called free radicals, are unstable molecules that contain oxygen and that easily react with other molecules in a cell; a build-up of these in cells may cause damage to dna, rna, and proteins, and may cause This cationic lipid-like structure, with clearly segregated hydrophobic and charged amine moieties, could explain this self-assembly behavior with DOPE, in contrast to loperamide, verapamil and epinastine. Cationic nanocarriers are reported to induce cell necrosis, especially in the lungs upon systemic administration. CAT level was significantly lower in group 2 than in groups 3- 6. . INTRODUCTION. "And we knew we couldn't use permanently positively charged lipids because they are so damn toxic." Those lipids would rip cell membranes apart, he adds. The two products provided that are made from Cationic PEGylated Lipids provided here consist of either DSPC:Chol:DSTAP:DSPE-PEG(2000 . aggregation phenomena in whole There is evidence that LNPs used in many preclinical studies are highly inflammatory (in mice), according to Ndeupen et al. Conclusions. However, despite having been widely used for more than 10 years for in vitro transfection applications, the use of these molecules has not yet been translated to application in humans because of low in vivo transfection efficiencies and toxicity issues. The most commonly used cationic liposomes utilized for the delivery of nucleic acids include DOTMA, DOTAP and DC-Chol. Cationic lipid-mediated delivery is a fast, simple, and reproducible means for easily introducing DNA, RNA, siRNA, or oligonucleotides into eukaryotic cells. They are especially useful for the DNA-dependent transformation of cells. Cationic lipids and polymers, the most important non-viral vectors, have many advantages over viral ones as non-immunogenic, easy to produce and not oncogenic. For the new lipid, the transfection efficiency remains essentially unchanged, while it drops quickly for DOTAP. Abstract Two novel cationic lipids based on protonated cyclen and steroid (cholesterol or diosgenin) moieties with carbamate linkage have been designed and synthesized for gene delivery. It is reported that some LNPs. J. Cationic lipids are non-viral vectors, which resemble traditional pharmaceuticals, display little immunogenicity, and have no potential for viral infection. The cell . However, toxicity and low transfection efficiency are two barriers limiting the clinical applications of cationic lipids. USA 84:7413-7417; Eppstein, D. et al., U.S. Pat. In this paper, the safety and toxicity of this formulation is described in two A solution came from new lipids that were charged only under certain conditions. Release. 2015). The lipoplexes formed from both cationic lipids were found to have low cytotoxicity in three cell lines even at high N/P ratios (see plot for H460 cells). We report the syntheses of novel cationic lipids comprised of cholesteryl-moieties linked to guanidinium functional groups, and also cationic lipids comprising a dialkylglycylamide moiety conjugated with a polyamine or a guanidinium functional group. The research shows that many derivatives of cholesterol which contain tertiary or quaternary nitrogen headgroups can inhibit PKC activity. It allows the highly efficient transfection of a broad range of cell types, including adherent, suspension, and insect cells, as well as primary cultures. Partial blocking of the surface charge of cationic nanocarriers might . In recent year, cationic liposomes have gained a lot of attention for siRNA delivery. Various cationic lipids have been synthesized and tested for use in nonviral gene delivery (Filion & Phillips, 1997). In some embodiments, a cationic lipid may be used to facilitate a charge-charge interaction with nucleic acids. It allows the highly efficient transfection of a broad range of cell types, including adherent, suspension, and insect cells, as well as primary cultures. Because all cationic lipids exhibit some level of cytotoxicity in vitro, we assessed the safety profile of one such cationic lipid, GL-67, following administration into the lungs of BALB/c mice. 14-16 cationic lipids are positively charged amphiphiles consisting of three basic chemical functional domains: a The mRNA then binds to ribosomesthe cell's little protein factoriesand induces the synthesis of the TLDR. 4,897,355.) back however, cationic lipids still have the problem of toxicity, as they can activate several cellular pathways like pro-apoptotic and pro-inflammatory cascades, which has become one of the main bottlenecks for their applications. Studying higher proportions of chol is necessary as it has been indicated that as chol content increases, cationic lipoplexes show less toxicity, more stability in serum conditions, and higher transfection ability ( 20, 21 ). There are five major structural classes of ionizable cationic lipids that have varying effects on RNA delivery and toxicity: unsaturated, multi-tail, branched-tail, polymeric, and biodegradable ionizable lipids. For usage as gene transfer vectors, cationic lipids are usually combined with neutral lipids such as dioleoylphosphatidylethanolamine (DOPE) or cholesterol (Chol). Cationic liposomes are made up of cationic lipids with two hydrophobic aliphatic long chains and positively charged functions in their head groups. This is of great importance for potential applications, since the cationic lipid is the more expensive compound and the toxicity of cationic lipids is a known problem in clinical applications. IGROV-1 and HeLa) both in the presence or absence of serum. Since the first studies were conducted, hundreds of cationic lipids have been synthesized as candidates for non-viral gene delivery and a few entered clinical trials. . First reported liposomes by Felgner was N- (1- (2,3-dioleyloxy)propyl)-N,N,N . So far, various cationic lipids have been developed with various transfection efficiencies and cellular toxicities. In fact, cationic liposome (CL) based vectors are among the prevalent synthetic carriers of . Here we explore the use of lipoaminoacids (LAAs) as cationic IL counterions that degrade or digest in vivo to non-toxic components. "There are no cationic lipids in nature," Cullis says. They hold the promise to replace viral vectors to be used in clinic. Despite this, intracellular barriers as endosomal escape and cytosolic delivery of siRNA still represent a challeng, as well as the cytotoxicity due to cationic lipids. These cationic-anionic lipid pairs drive the transition from the bilayer structure to the . significantly reduced the toxicity of the DDA liposomal . What 2006; 114:100-109 . ~80% endothelial gene knockdown in the lungs was achieved by 7C1 without significant toxicity 21, . The cationic lipids can become cytotoxic by interacting with critical enzymes such as PKC. The cationic/ionizable lipids, complex with the negatively charged mRNA molecules and enable the exit of the mRNA from the endosome to the cytosol for translation. this requirement cating that when the cationic lipids exceed has not been previously considered because a certain safe threshold (3.33%), this affects corresponding author. Typically, the nal concentration of lipid in chloroform was 20 mM. There is a real possibility that cationic lipids will accumulate in the ovaries. However, the toxicity is still an obstacle to the application of non-viral vectors to gene therapy. The release of damage-associated molecular patterns, such as mitochondrial DNA from the injured cell may result in the inflammatory toxicity of the nanocarrier, which has largely limited its clinical application. activity equivalent to, and toxicity significantly lower than, Lipofectin [38]. Although the toxicity of cationic lipids in vitro is well documented, to our knowledge, this is the first report detailing toxicity associated with their use in vivo, and highlights considerations with regard to the use of these reagents in human clinical studies. Cellular toxicity of CLs was assessed in HepG2 cells using the CCK-8 assay as described previously (Ko et al. of the lack of a suitable technique to study the cell viability by different mechanisms that e-mail: [email protected] (k. buyens). In this report, a novel cationic lipid, dimyristyloxypropyl-3-dimethyl-hydroxyethyl ammonium (DMRIE), has been substituted into the DNA-liposome complex with dioleoyl phosphatidylethanolamine (DOPE), which both improves transfection efficiencies and allows increased doses of DNA to be delivered in vivo. Cationic lipids and polymers, the most important non-viral vectors, have many advantages over viral ones as non-immunogenic, easy to produce and not oncogenic. Cationic lipids have been successfully employed as vectors for gene transfer in lung grafts, yet those lipid vectors have potential toxicity. One of the major disadvantages of synthetic non-viral cationic lipid-based systems is toxicity. Several strategies were adopted such as surface modification of polyethylene glycol (PEG) to . "There was a group of chemists put on this right away to build novel cationic lipids," says Ciaramella, the former head of infectious diseases at Moderna. However, the toxicity is still an obstacle to the application of non-viral vectors to gene therapy. The choice of lipid is predominantly between either a two hydrocarbon chain or a cholesterol moiety. 2021. The cationic lipids, i.e., dimethyldioctadecylammonium (DDA), 1,2-dioctadecanoyl-sn-glycero-3-phosphocholine (DSPC), and D--tocopherol polyethylene glycol 1000 (TPGS) can form blank liposomes, which can incorporate influenza antigens to produce an influenza vaccine (DDA-DSPC-TPGS). Cationic solid lipid nanoparticles (cSLNs) represent one alternative to deliver poorly water-soluble drugs and oligonucleotides (e.g., DNA, siRNA, miRNA) based on well-known biocompatible and safe excipients. Ionizable cationic lipids available from Cayman and their pK a values can be found at the end of this article. Cationic lipid/DNA complexes manufactured by microfluidics and bulk self-assembly exhibit different transfection behavior. . 2005). The pulmonary toxicity of cationic lipid is dose-dependent. To prepare cationic liposomes containing pacli-taxel, known concentrations of drug dissolved in methanol (10 mg/mL) were mixed with the required amount of lipid dissolved in chloroform. The present invention is directed to new cationic lipids and intermediates in their synthesis that are useful for transfecting nucleic acids or peptides into prokaryotic or eukaryotic cells. Objective: This paper reviews the lipid structure of cationic lipids, their structure-activity relationship, toxicity and transfection View on Taylor & Francis Save to Library Cationic lipid transfection reagents. January 7, 2022: Dr. Michael Palmer from the University of Waterloo offers expert insights into the connection between mRNA vaccine technology and Cationic Lipid Toxicity.In this brief video, the viewer is provided with a simple but critically important understanding of Cationic Lipid Toxicity and the relevance of this theme when it comes to short-term and long-term personal health. The lipid that is used in LNP for the COVID-19 mRNA vaccines, are cationic lipid. Also disclosed are lipids useful both for the delivery of macromolecules and also useful as intermediates for making other such lipids. 30, 31 Most of our previous work has employed . To address these issues, we developed four liposomal formulations, composed of two different cationic lipids (DOTAP and DC-Cholesterol) and . During the experiment, HepG2 . This binding force plays an important role in the therapeutic efficacy of gene therapy. The lipid . Hence, we investigate the toxicity of cSLNs formulation . Journal of medicinal chemistry. The mechanisms of toxicity of cationic lipid are thought to be alterations of the net charge of cell membranes and adverse effects on the activity of ion channels, membrane receptors, and enzymes ( 2 ). Cationic Liposomes for Gene Delivery: Novel Cationic Lipids and Enhancement by Proteins and Peptides . Methods Although the transferring efficacy of CLs is substantially lower than that of viral . They demonstrated that mRNA is located in the water column surrounded by cationic lipids . Advantages of cationic lipid-mediated transfection include high efficiency, ability to transfect a wide range of cell types, reproducibility, low toxicity, and simplicity. Toxicity of cationic lipids and cationic polymers in gene delivery. However, the toxicity is still an obstacle to the application of non-viral vectors to gene therapy. Most of the cationic lipid-based nanoparticles are prone to evoke toxicity because of using cationic lipids and easy to aggregate with serum components due to extra positive surface charges (Gebeyehu et al., 1994; Scheule et al., 1997; Lv et al., 2006). Cationic liposomes are the most common lipid-based nanocarrier used for siRNA delivery. 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