Messenger RNA (mRNA)-based vaccines have shown great potential to activate antitumor immune response for cancer therapy. Numerous unsolved technical difficulties in successful in vivo delivery of in vitro transcribed (IVT) mRNA, such as transfection efficiency, have greatly hindered the applications of IVT-mRNA in clinical development. As one of the lipid-based vectors, lipoplexes have recently been the subject of intensive investigations to understand the parameters governing the efficiency of transfection.
Equipped with world-leading technology platforms and professional scientific staff in the development of the drug delivery system, Creative Biolabs is committed to providing high-quality lipoplex-based delivery systems to efficiently facilitate targeted in vivo delivery of IVT-mRNA in safe manner.
mRNA is frequently encapsulated with positively charged liposomes to form lipoplexes. Lipoplexes are self-assembling nanosystems and their formation entails a multistep mechanism as revealed by detailed studies employing monolayers, atomic force microscopy and cryo-electron microscopy. In a millisecond first step, electrostatic interactions between phosphate and the positively charged amine headgroup of the cationic lipid occur. The cellular uptake is mainly mediated by either lipoplex endocytosis or endocytosis-like mechanisms. Moreover, lipoplex delivery systems have substantial advantages like easy synthesis, low batch variability, scalability, and biocompatibility. It has also been noted that drug delivery systems in the nano-range enhance the pharmacokinetic availability of encapsulated drugs.
IVT-mRNA has been applied as a therapeutic molecule in the field of cancer therapy and biomedical research studies. mRNA-based therapy has shown several advantages but its further application is largely restricted by delivery method.
Researches have developed a liposome-protamine lipoplex (CLPP) to deliver mRNA encoding survivin-T34A gene, resulting in a novel core-shell structured nanoparticle formulation (CLPP/mSur-T34A). This CLPP/mSur-T34A formulation displayed high mRNA transfecting and expression efficiency on C26 tumor cells through lipid rafts-mediated endocytosis. CLPP/mSur-T34A mRNA formulation showed obvious therapeutic effects on various models of C26 colon cancer both in vitro and in vivo. The results indicated the high delivery capacity of lipoplex and further suggested that CLPP/mSur-T34A mRNA formulation can be a potential candidate for colon cancer therapy.
Another team reported a lipoplex-based mRNA nanovaccine which can greatly improve mRNA delivery in antigen-presenting cells, activate T cell response, and induce robust antitumor immunity. This lipoplex-based formulation is composed by cationic polymer/lipid(poly(amidoamine)/alkylene oxide), DSPE-PEG2000, and epitope-encoding mRNA. With appropriate ratio, the optimized nanovaccine demonstrated effective mRNA delivery in dendritic cells, and strongly active antigen-specific immune response in vitro and in vivo.
Fig.1 Negatively charged IVT-mRNA spontaneously interact with cationic lipids to form lipoplexes. (Guan, 2017)
At Creative Biolabs, liposomes are usually composed of 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) and 1,2-distearoyl-sn-glycero3-phosphoethanol-amine-N-[biotinyl(polyethylene glycol)-2000] (DSPE-PEG-2000-biotin). With years of experience in nanotechnology and material science, our scientists are more than happy to share our cutting-edge technologies and experience in the development of lipoplex-based delivery system for the transfection of therapeutic mRNA. Moreover, we also provide a comprehensive analysis of mRNA-lipoplexes, including transfection efficiency, duration of protein expression, immunogenicity, hemocompatibility, and storage stability.
The diversity and flexibility of lipoplex-based delivery systems highlight the recent progress of IVT-mRNA-based therapy. To assist our clients in the development of lipoplex-based delivery systems, Creative Biolabs is proud to offer the most comprehensive services. For more detailed information, please feel free to contact us or directly send us an inquiry.