Minicircle Sys tem as An Innovative Approach forGeneration of Functional CAR T-Cell

Objective: Chimeric antigen receptors (CAR) are receptors thatgives T-cells a new ability to target a specific cancer antigen. Ithas been shown that this s trategy is an efficient and safe approachfor treatment of hematological and some solid cancers.On the other hand, the minicircle as a nonviral vector has theability to high level expression of the transgene without adverseeffects compared to the viral vectors. In minicircle, prokaryoticsequences are deleted from the parental plasmid and the vectoris a smaller in size with a long duration of expression, and saferdue to the absence of bacterial sequences. These characteris ticshave made the minicircle as a promising vector for biomedicalapplications. In the present s tudy, we applied minicircle as asafe vector to generate specific and functional CAR T-cells.Materials and Methods: In this s tudy, a coding sequence ofthe third generation of CAR was cons tructed, and cloned intoa replicative minicircle. This CAR encoding minicircle (CARMC)was introduced into T-cells from peripheral blood by electroporation.Then, resulted CAR T-cells were co-cultured withthe cancer cells with the specific antigen. Functionality of CAR T-cells were tes ted in vitro. Generated.Results: The results showed that CAR T-cells produced byCAR-MC, specifically targeted the cancer cells with s trongcytotoxic effects compared to negative cells. Additionally,these CAR T-cells also exhibited the ability to secret a significantlyhigher level of inflammatory cytokines compared to thecontrol T-cells.Conclusion: Overall, the present s tudy showed that the CARMChas the capability of improving antitumor activity of theT-cells. In fact, minicircle encoding a functional CAR receptorenabled the T-cells to exclusively target and des troy themalignant cells

کلیدواژه ها:

Cancer Immunotherapy ، CAR T-Cell ، CD۱۹ ، Minicircle