Multiplex genome-edited T-cell manufacturing platform for “off-the-shelf” adoptive T-cell immunotherapies

L Poirot, B Philip, C Schiffer-Mannioui, D Le Clerre… - Cancer research, 2015 - AACR
L Poirot, B Philip, C Schiffer-Mannioui, D Le Clerre, I Chion-Sotinel, S Derniame, P Potrel…
Cancer research, 2015AACR
Adoptive immunotherapy using autologous T cells endowed with chimeric antigen receptors
(CAR) 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 …
Abstract
Adoptive immunotherapy using autologous T cells endowed with chimeric antigen receptors (CAR) 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 αβ 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 biologic immunopharmaceuticals. Cancer Res; 75(18); 3853–64. ©2015 AACR.
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