Open AccessmRNA-based CAR T-cells manufactured by miniaturized two-step electroporation produce selective cytotoxicity toward target cancer cells
Author(s) -
Vidura Jayasooriya,
Beth Ringwelski,
Glenn Dorsam,
Dharmakeerthi Nawarathna
Publication year - 2021
Publication title -
lab on a chip
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.064
H-Index - 210
eISSN - 1473-0197
pISSN - 1473-0189
DOI - 10.1039/d1lc00219h
Subject(s) - electroporation , chimeric antigen receptor , cytotoxicity , cytotoxic t cell , transfection , cancer cell , microbiology and biotechnology , biology , messenger rna , immunotherapy , cell culture , in vitro , immunology , cancer , biochemistry , immune system , gene , genetics
There is a growing interest for viral vector-free chimeric antigen receptor (CAR) T-cells due to its ability to kill cancer cells without adverse side effects. A potential avenue for manufacturing viral-vector free CAR T-cells is to utilize mRNA electroporation. One of the major concerns with mRNA electroporated CAR T-cells is the shorter cytotoxic lifespan of a few days, which is insufficient or not ideal for therapy. To better understand this issue and develop a potential solution, this study focused on examining the translation of electroporated mRNA to CAR molecules, time dependent degradation of CAR molecules and cytotoxicity produced by CAR T-cells on cancer cells. It was found that the initial expression of CAR molecules dictates the cytotoxicity. Initial CAR expression could be controlled by the experimental parameters such as electroporation time and mRNA concentration in the electroporation buffer. Experiments were carried out using a novel two-step electroporation that allows for controlled and uniform transfection of T-cells. These technical advancements and subsequent findings could provide a viable path for producing CAR T-cells with longer cytotoxic lifespans.