Adoptive cell therapy (ACT) with retargeted T cells has produced remarkable clinical responses against cancer， but also serious toxicity. Telomerase is overexpressed in most cancers， but also expressed in some normal cells， raising safety concerns. We hypothesize that ACT with T-helper cell receptors may overcome tumour tolerance， mobilize host immune cells and induce epitope spreading， with limited toxicity. From long term survivors after cancer vaccination， we have isolated telomerase-specific T cell receptors (TCRs) from T-helper cells. Herein， we report the development of transient retargeting of T cells with mRNA-based TCRs. This strategy allows for safer clinical testing and meaningful dose ... More
Adoptive cell therapy (ACT) with retargeted T cells has produced remarkable clinical responses against cancer， but also serious toxicity. Telomerase is overexpressed in most cancers， but also expressed in some normal cells， raising safety concerns. We hypothesize that ACT with T-helper cell receptors may overcome tumour tolerance， mobilize host immune cells and induce epitope spreading， with limited toxicity. From long term survivors after cancer vaccination， we have isolated telomerase-specific T cell receptors (TCRs) from T-helper cells. Herein， we report the development of transient retargeting of T cells with mRNA-based TCRs. This strategy allows for safer clinical testing and meaningful dose escalation. DP4 is the most common HLA molecule. We cloned two telomerase-specific， DP4-restricted TCRs into the mRNA expression vector pCIpA102， together with the sorter/marker/suicide gene RQR8. Donor T cells were electroporated with mRNA encoding TCR_RQR8. The results showed that both TCR_RQR8 constructs were expressed in >90% of T cells. The transfected T cells specifically recognized the relevant peptide， as well as naturally processed epitopes from a 177aa telomerase protein fragment， and remained functional for six days. A polyfunctional and Th1-like cytokine profile was observed. The TCRs were functional in both CD4+and CD8+recipient T cells， even though DP4-restricted. The findings demonstrate that the cloned TCRs confer recipient T cells with the desired telomerase-specificity and functionality. Preclinical experiments may provide limited information on the efficacy and toxicity of T-helper TCRs， as these mobilize the host immune system. We therefore intend to use the mRNA-based TCRs for a first-in-man trial.
