Abstract
10027 Background: Highly targeted immunotherapy with adoptively transferred chimeric antigen receptor (CAR) T cells has shown considerable efficacy against some refractory childhood malignancies. Downregulation/mutation of targeted antigen can however create antigen loss escape variants. Multispecific T-cell approach could therefore further improve their therapeutic efficacy. Methods: We created a novel bispecific CAR that incorporates two extracellular antigen-recognition domains; one for HER2 (from HER2 monoclonal antibody FRP5) and the other for IL-13Rα2 (a mutated IL-13 molecule), in tandem (TanCAR). TanCAR was rationally designed by in silico modeling. Intracellular domain consists of a CD28/zeta-signaling chain. TanCAR encoding DNA was then synthesized and force expressed on T cells using retroviral system. Surface expression of extracellular domain of the TanCAR in its entirety was confirmed using HER2/IL-13Rα2 specific methods on flowcytometry. Functionality of TanCAR T cells was evaluated using s...
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