Research Of MRNA Vaccines Against The Tumor-specific Antigen MAGE-C2

There are several approaches to tumor immunotherapy,and it has been found that effective antigen presentation and immune checkpoint inhibition are required for successful immune activation.Tumor vaccines can enhance tumor antigen presentation,and mRNA vaccines have the advantages of being highly effective and safe compared to conventional vaccines.However,its application has been limited by technical problems such as unstable in vivo,easy degradation,low transfection and translation efficiency,etc.Its stability and translation efficiency can be improved by structural modification and the use of liposomal nanoparticles,etc.Since 2019,with the SARS-COV2 mRNA vaccine approval,tumor mRNA vaccines have also gained rapid progress.The aim of this project is to prepare and initially test anti-tumor mRNA vaccines to gain experience for subsequent development.Melanoma-associated antigen(MAGE-C2),a tumor-specific protein that is not expressed in normal human tissues(except testicular spermatogonia)but only in malignant tumor tissues,was chosen to design the mRNA vaccine.MAGE-C2 is involved in tumorigenesis,development and metastasis,and is associated with poor prognosis,making it a good target for tumor immunotherapy.A recombinant protein vaccine against MAGE-C2 has been studied,but the therapeutic effect is still unsatisfactory.In this study,we used Themofisher online tool to optimize the codon of MAGE-C2 in order to improve its translation efficiency and stability,followed by in vitro transcription,capping and tailing reactions to obtain mRNA.Lipid nano-particles(LNPs)bodies containing mRNA were prepared using microfluidic technology.The particle size and shape were assessed,and the encapsulated mRNA was transfected into B16 cells.The expression of MAGE-C2 was detected by Western blot and cellular immunohistochemistry.