NY-ESO-1 combined with MPLA vaccine

As a model TCR, we selected a 1G4 clone that reacts with NY-ESO-1-derived peptide in an HLA-A*02-restricted manner... We have successfully designed Platinum TALEN mRNA pairs targeting TRAC and TRBC which facilitate the production of genome-edited human primary T cells. We also developed an electroporation and expansion culture protocol that enables us to produce viable genome-edited 1G4-transduced T cells at a large cell dose equivalent to a clinical scale. Collectively, these results suggest that targeted orthotopic knock-in of antigen-specific TCR-encoding ssDNA into the TCR locus by the use of Platinum TALEN is a promising strategy that can be applied for the clinical manufacturing of therapeutic TCR-T cells.

Primary T cells from two donors were transduced with lentivirus containing a NY-ESO-1 TCR as well as a genome-wide sgRNA library and subsequently CRISPR edited using Cas9-RNP electroporation...The template for the B Cell Maturation Antigen (BCMA)-specific CAR bb2121 was then delivered to the TRAC locus via CRISPR/Cas9-RNP and adeno-associated virus (AAV)...We found a significant advantage in tumor killing of CUL5 ablated T cells compared to control cells in this ex vivo setting, suggesting an improved functional persistence. In summary, our findings identify the CUL5 complex as a novel potential therapeutic target to improve T cell persistence and counteract T cell exhaustion in order to improve the efficacy of adaptive cell immunotherapies for the treatment of BCMA+ hematological malignancies.

TCR-engineered T cells derived from HLA-A*0201-typed NSCLC patients co-cultured with patient-derived cancer cells expressing NY-ESO-1 and analysed by flow cytometry for their killing activity, demonstrated that TCR engineering enabled T cells to recognize and kill NSCLC cells. These data strongly encourage the application of CRISPR-mediated non-viral TCR editing to generate tumor-specific T cells able to kill NSCLC cells, contributing to the preclinical validation of adoptive T cell therapy with CRISPR-mediated engineered T cells for the treatment of lung cancer.