title: Multiplex Genome-Edited T-cell Manufacturing Platform for "Off-the-Shelf" Adoptive T-cell Immunotherapies.
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
authors.affiliation: Cellectis, Paris, France.
authors.forename: Laurent
authors.initials: L
authors.lastname: Poirot
authors.affiliation: Department of Haematology, UCL Cancer Institute, University College London, London, United Kingdom.
authors.forename: Brian
authors.initials: B
authors.lastname: Philip
authors.affiliation: Cellectis, Paris, France.
authors.forename: Cécile
authors.initials: C
authors.lastname: Schiffer-Mannioui
authors.affiliation: Cellectis, Paris, France.
authors.forename: Diane
authors.initials: D
authors.lastname: Le Clerre
authors.affiliation: Cellectis, Paris, France.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Chion-Sotinel
authors.affiliation: Cellectis, Paris, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Derniame
authors.affiliation: Cellectis, Paris, France.
authors.forename: Pierrick
authors.initials: P
authors.lastname: Potrel
authors.affiliation: Cellectis, Paris, France.
authors.forename: Cécile
authors.initials: C
authors.lastname: Bas
authors.affiliation: Cellectis, Paris, France.
authors.forename: Laetitia
authors.initials: L
authors.lastname: Lemaire
authors.affiliation: Cellectis, Paris, France.
authors.forename: Roman
authors.initials: R
authors.lastname: Galetto
authors.affiliation: Cellectis, Paris, France.
authors.forename: Céline
authors.initials: C
authors.lastname: Lebuhotel
authors.affiliation: Cellectis, Paris, France.
authors.forename: Justin
authors.initials: J
authors.lastname: Eyquem
authors.affiliation: Department of Haematology, UCL Cancer Institute, University College London, London, United Kingdom.
authors.forename: Gordon Weng-Kit
authors.initials: GW
authors.lastname: Cheung
authors.affiliation: Cellectis, Paris, France.
authors.forename: Aymeric
authors.initials: A
authors.lastname: Duclert
authors.affiliation: Cellectis, Paris, France.
authors.forename: Agnès
authors.initials: A
authors.lastname: Gouble
authors.affiliation: Cellectis, Paris, France.
authors.forename: Sylvain
authors.initials: S
authors.lastname: Arnould
authors.affiliation: Department of Haematology, UCL Cancer Institute, University College London, London, United Kingdom.
authors.forename: Karl
authors.initials: K
authors.lastname: Peggs
authors.affiliation: Department of Haematology, UCL Cancer Institute, University College London, London, United Kingdom.
authors.forename: Martin
authors.initials: M
authors.lastname: Pule
authors.affiliation: Department of Pediatrics, University of Washington, Seattle Children's Research Institute, Seattle, Washington.
authors.forename: Andrew M
authors.initials: AM
authors.lastname: Scharenberg
authors.affiliation: Cellectis, Paris, France. smith@cellectis.com.
authors.forename: Julianne
authors.initials: J
authors.lastname: Smith
mesh_terms: D000074323:Alemtuzumab; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D061067:Antibodies, Monoclonal, Humanized / Q000494:pharmacology; D015703:Antigens, CD / Q000235:genetics; D018941:Antigens, CD19 / Q000276:immunology; D000951:Antigens, Neoplasm / Q000235:genetics / Q000276:immunology; D001483:Base Sequence; D000074301:CD52 Antigen; D003602:Cytotoxicity, Immunologic; D004351:Drug Resistance; D055786:Gene Knockout Techniques; D006023:Glycoproteins / Q000172:deficiency / Q000235:genetics; D006086:Graft vs Host Disease / Q000517:prevention & control; D006801:Humans; D016219:Immunotherapy, Adoptive; D008213:Lymphocyte Activation; D008223:Lymphoma / Q000628:therapy; D051379:Mice; D008817:Mice, Mutant Strains; D008969:Molecular Sequence Data; D012333:RNA, Messenger; D011948:Receptors, Antigen, T-Cell / Q000235:genetics / Q000276:immunology; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000172:deficiency / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D013601:T-Lymphocytes / Q000187:drug effects / Q000276:immunology / Q000637:transplantation*; D014162:Transfection; D023041:Xenograft Model Antitumor Assays
chemical_list: D000911:Antibodies, Monoclonal; D061067:Antibodies, Monoclonal, Humanized; D015703:Antigens, CD; D018941:Antigens, CD19; D000951:Antigens, Neoplasm; D000074301:CD52 Antigen; C000613862:CD52 protein, human; D006023:Glycoproteins; D012333:RNA, Messenger; D011948:Receptors, Antigen, T-Cell; D016693:Receptors, Antigen, T-Cell, alpha-beta; D011993:Recombinant Fusion Proteins; D000074323:Alemtuzumab