We don't have a Cellectis thread, but this is relevant to juno......
cancerres.aacrjournals.org
Multiplex genome edited T-cell manufacturing platform for "off-the-shelf" adoptive T-cell immunotherapies
Laurent Poirot 1, Brian Philip 2, Cécile Schiffer-Mannioui 1, Diane Le Clerre 1, Isabelle Chion-Sotinel 1, Sophie Derniame 1, Cécile Bas 1, Pierrick Potrel 1, Laetitia Lemaire 1, Aymeric Duclert 1, Roman Galetto 1, Céline Lebuhotel 1, Justin Eyquem 1, Gordon W Cheung 2, Agnès Gouble 1, Sylvain Arnould 1, Karl S. Peggs 2, Martin Pule 2, Andrew Scharenberg 3, and Julianne Smith 1, *
1CAR development group, Cellectis2Department of Haematology, UCL Cancer Institute3Department of Pediatrics, Seattle Children's Research Institute, University of Washington ?* Corresponding Author:
Julianne Smith, CAR development group, Cellectis, 8 rue de la croix Jarry, Paris, 75013, France smith@cellectis.com
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. |
