Multiplex genome edited T-cell manufacturing platform for "off-the-shelf" adoptive T-cell immunotherapies
Menée in vitro et in vivo, cette étude évalue la faisabilité d'une technologie d'ingéniérie des génomes (TALEN) pour fabriquer des immunothérapies adoptives à base de lymphocytes T allogènes modifiés pour exprimer un récepteur antigénique chimérique
Adoptive immunotherapy using autologous T-cells endowed with chimeric antigen receptors (CARs) has emerged as a powerful means of treating cancer. However, a limitation of this approach is that autologous CAR T-cells must be generated on a custom-made basis. Here we show that electroporation of transcription activator-like effector nuclease (TALEN) mRNA allows highly-efficient multiplex gene editing in primary human T-cells. We use this TALEN-mediated editing approach to develop a process for the large-scale manufacturing of T-cells deficient in expression of both their T-cell receptor (TCR) and CD52, a protein targeted by alemtuzumab, a chemotherapeutic agent. Functionally, T-cells manufactured with this process do not mediate graft-versus-host reactions, and are rendered resistant to destruction by alemtuzumab. These characteristics enable the administration of alemtuzumab concurrently or prior to engineered T-cells, supporting their engraftment. Furthermore, endowing the TALEN-engineered cells with a CD19 CAR led to efficient destruction of CD19+ tumor targets even in the presence of the chemotherapeutic agent. These results demonstrate the applicability of TALEN-mediated genome editing to a scalable process, which enables the manufacturing of third-party CAR T-cell immunotherapies against arbitrary targets. As such, CAR T-cell immunotherapies can therefore be used in an "off-the-shelf" manner akin to other biological immunopharmaceuticals.