Multiplex Genome-Edited T-cell Manufacturing Platform for "Off-the-Shelf" Adoptive T-cell Immunotherapies

作者：Poirot, Laurent; Philip, Brian; Schiffer-Mannioui, Cecile; Le Clerre, Diane; Chion-Sotinel, Isabelle; Derniame, Sophie; Potrel, Pierrick; Bas, Cecile; Lemaire, Laetitia; Galetto, Roman; Lebuhotel, Celine; Eyquem, Justin; Cheung, Gordon Weng-Kit; Duclert, Aymeric; Gouble, Agnes; Arnould, Sylvain; Peggs, Karl; Pule, Martin; Scharenberg, Andrew M.; Smith, Julianne^*

来源：Cancer Research, 2015, 75(18): 3853-3864.

DOI：10.1158/0008-5472.CAN-14-3321

• CHIMERIC-ANTIGEN-RECEPTOR
• IN-VIVO
• CANCER
• ANTIBODIES
• DEPLETION
• SPECIFICITY
• PERSISTENCE
• ALEMTUZUMAB
• EXPRESSION
• THERAPY

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 ab 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 CARled 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.

• 出版日期2015-9-15