Ricerc@Sapienza

On March 2020, the World Health Organization (WHO) declared the coronavirus disease a global pandemic. The immense impact of the pandemic on the global health and economics led to a massive resource¿s involvement in development of vaccine against SARS-Cov2. Thanks to this, big companies were able to produce efficiently and quickly, mRNA vaccines encapsulated in lipid nanoparticles (LNPs) which were never been approved before. Despite mRNA vaccines safety and effectiveness there are limits in their use. In fact, they need a cold chain to be maintained and delivered leading to logistical problems in view of mass vaccination. Furthermore, mRNA vaccines are quite expensive in respect to other most traditional vaccines. Considering this, DNA vaccines may represent a good alternative to mRNA vaccines as they do not require extremely low temperature for the storage and are relatively inexpensive. Importantly, the plasmid DNA containing the viral protein sequence can be easily modified to make the vaccine specific for the viral variants. pDNA needs to be conveyed into the target cells and to reach the nucleus to be transcribed and then translated into protein. Electroporation is the most diffused way to achieve the pDNA internalization however, it is an invasive procedure that may cause several side effects. For these reasons, the use of LNPs is preferable. Our group aims to produce a library of lipid-nanoparticles through microfluidic mixing technique, which offer the advantage to yield highly reproducible and low size nanoparticles. Different multicomponent lipid formulations will be characterized and tested in vitro on keratinocyte cell lines to evaluate the transfection efficiency (TE) and toxicity. Computational analysis of the LNPs features will allow their further optimization. Finally, the most performing LNPs will be functionalized using specific proteins improving the targeting to keratinocytes and dendritic cells, in view of subcutaneous vaccine administration.