ABSTRACT
Clinical trials with direct administration of synthetic mRNAs encoding tumor antigens demonstrated safety and induction of tumor-specific immune responses. Their proper delivery to dendritic cells (DCs) requires their protection against RNase degradation and more specificity for dose reduction. Lipid-Polymer-RNA lipopolyplexes (LPR) are attractive mRNA delivery systems and their equipment with mannose containing glycolipid, specific of endocytic receptors present on the membrane of DCs is a valuable strategy. In this present work, we evaluated the capacity of LPR functionalized with a tri-antenna of α-d-mannopyranoside (triMN-LPR) concerning (i) their binding to CD209/DC-SIGN and CD207/Langerin expressing cell lines, human and mouse DCs and other hematopoietic cell populations, (ii) the nature of induced immune response after in vivo immunization and (iii) their therapeutic anti-cancer vaccine efficiency. We demonstrated that triMN-LPR provided high induction of a local inflammatory response two days after intradermal injection to C57BL/6 mice, followed by the recruitment and activation of DCs in the corresponding draining lymph nodes. This was associated with skin production of CCR7 and CXCR4 at vaccination sites driving DC migration. High number of E7-specific T cells was detected after E7-encoded mRNA triMN-LPR vaccination. When evaluated in three therapeutic pre-clinical murine tumor models such as E7-expressing TC1 cells, OVA-expressing EG7 cells and MART-1-expressing B16F0 cells, triMN-LPR carrying mRNA encoding the respective antigens significantly exert curative responses in mice vaccinated seven days after initial tumor inoculation. These results provide evidence that triMN-LPR give rise to an efficient stimulatory immune response allowing for therapeutic anti-cancer vaccination in mice. This mRNA formulation should be considered for anti-cancer vaccination in Humans.
Subject(s)
Cancer Vaccines/administration & dosage , Dendritic Cells/immunology , Neoplasms/therapy , RNA, Messenger/administration & dosage , Animals , Antigens, Neoplasm/immunology , Cancer Vaccines/immunology , Cell Line, Tumor , Cell Movement/immunology , Female , Humans , Injections, Intradermal , Lipids/chemistry , Lymph Nodes/immunology , Mannose/chemistry , Melanoma, Experimental/immunology , Melanoma, Experimental/therapy , Mice , Mice, Inbred C57BL , Neoplasms/immunology , Ovalbumin/immunology , VaccinationABSTRACT
The interaction between glycine betaine-based cationic surfactant and algal polysaccharide κ-carrageenan was studied by investigating the dilution effect of the surfactant/polymer assemblies driven by electrostatic interactions. Two aqueous solutions of cationic surfactant and κ-carrageenan at two molar ratios (3.5 and 0.8) diluted with factors of 5 and 10 times, were tested by various analytical methods including a multiscale observation by Transmission Electron Microscopy (TEM) and Laser Scanning Confocal Microscopy (LSCM) to understand the solution behavior of surfactant and oppositely charged polymer at both nano- and micrometer scale. Raman spectroscopy as well as confocal Raman imaging were applied to give Supplementary information about the surfactant/polysaccharide interactions and the distribution of assemblies. These analyses confirmed the formation of singular hybrid surfactant/polymer nano-, microobjects and they revealed the influence of dilution on the nanostructures. These results give an insight of the mechanism of the dilution effect on surfactant/polymer assemblies that could be valuable in pharmaceutical formulations, food and cosmetics fields.
ABSTRACT
Are the liquid crystalline properties of the materials of living systems important in biological structures, functions, diseases and treatments? There is a growing consciousness that the observed lyotropic, and often thermotropic liquid crystallinity, of many biological materials that possess key biological functionality might be more than curious coincidence. Rather, as the survival of living systems depends on the flexibility and reformability of structures, it seems more likely that it is the combination of softness and structure of the liquid-crystalline state that determines the functionality of biological materials. The richest sources of liquid crystals derived from living systems are found in cell membranes, of these glycolipids are a particularly important class of components. In this critical review, we will examine the relationship between chemical structure and the self-assembling and self-organising properties of glycolipids that ultimately lead to mesophase formation.
Subject(s)
Glycolipids/chemistry , Liquid Crystals/chemistry , Temperature , Glycolipids/chemical synthesis , Models, Molecular , Molecular Conformation , Particle Size , Surface PropertiesABSTRACT
Novel cationic liposomes, termed "archaeosomes", based on mixtures of neutral/cationic bilayer-forming lipids and archaeobacterial synthetic tetraether-type bipolar lipids show efficient in vitro gene transfection properties and represent a new approach for modulating the lipidic membrane fluidity of the complexes they form with DNA.
Subject(s)
Glyceryl Ethers/chemistry , Transfection/methods , Cations/chemistry , Cell Line, Tumor , DNA/genetics , Glyceryl Ethers/chemical synthesis , Humans , Liposomes , Molecular Structure , Plasmids/geneticsABSTRACT
New dissymmetrical neutral-cationic or anionic-cationic alpha,omega-diamido bolaamphiphiles have been synthesized in which the polar headgroups are derived from alginate and glycine betaine and which exhibit monolayer lipid membrane vesicles, large lamellae and rods.
Subject(s)
Alginates/chemistry , Membranes, Artificial , Oligosaccharides/chemistry , Surface-Active Agents/chemical synthesis , Molecular Conformation , Particle Size , Stereoisomerism , Surface-Active Agents/chemistryABSTRACT
The in vitro stability of archaeosomes made from novel synthetic membrane-spanning tetraether lipids was evaluated in conditions mimicking those of oral route application in terms of bile salts, serum and low pH..
Subject(s)
Drug Delivery Systems , Ether/chemistry , Lipids/chemistry , Administration, Oral , Lipids/chemical synthesis , Microscopy, Electron , Molecular StructureABSTRACT
A liquid chromatographic method has been developed for the analysis of polyglycerol fatty esters and fatty ethers which are non-ionic surfactants. Two methods were compared using either octadecyl silica or porous graphitic carbon. The octadecyl silica system with a hydroorganic mobile phase enables to compare the hydrophobic behavior of the compounds. The porous graphitic carbon enables the separation of the diastereoisomers. Detection of these non-UV-absorbing compounds was achieved by evaporative light scattering detection. Identification of isomers was performed by mass spectrometry.
Subject(s)
Chromatography, Liquid/methods , Esters/analysis , Ethers/analysis , Fatty Acids/chemistry , Glycerol/chemistry , Polymers/chemistry , Graphite , Light , Mass Spectrometry , Scattering, Radiation , Silicon DioxideABSTRACT
With the aim of developing new efficient agents for transfecting of eukaryotic cells we have designed and synthesized a novel family of cationic lipid vectors derived from glycine betaine. In this study we present three novel molecules differing by the length of their aliphatic chains (R=12,R=14,R=16). The lyotropic properties of these cationic lipids have been determined, and their transfection efficiency on different cell lines evaluated, using a luminescent assay. Two of these compounds, GB14 and GB12 are efficient in vitro experiments. Cytoxicity evaluation of these new molecules showed promising results with a low cytotoxicity, especially when co-lipids were included in the formulation. These compounds represent a new family of gene transfer vectors which display good transfection efficiency and low toxicity, possibly due to the natural properties of glycine betaine. These compounds have great potential for the future development of in vivo gene transfer protocols.
