Maria Michela D'Aloia1, Sara Caratelli2, Camilla Palumbo3, Simone Battella4, Roberto Arriga5, Davide Lauro5, Gabriella Palmieri4, Giuseppe Sconocchia6, Maurizio Alimandi1. 1. Department of Clinical and Molecular Medicine, Sapienza University of Rome, Rome, Italy. 2. Department of Biomedicine, Institute of Translational Pharmacology, CNR, Rome, Italy; Laboratory of Molecular Medicine, Department of Systems Medicine, Tor Vergata University, Rome, Italy. 3. Department of Clinical Sciences and Translational Medicine, Tor Vergata University, Rome, Italy. 4. Department of Experimental Medicine, Sapienza University of Rome, Rome, Italy. 5. Laboratory of Molecular Medicine, Department of Systems Medicine, Tor Vergata University, Rome, Italy. 6. Department of Biomedicine, Institute of Translational Pharmacology, CNR, Rome, Italy. Electronic address: giuseppe.sconocchia@cnr.it.
Abstract
BACKGROUND AIMS: Chimeric antigen receptors (CARs) designed for adoptive immunotherapy need to achieve two functions: antigen recognition and triggering of the lytic machinery of reprogrammed effector cells. Cytotoxic T cells have been engineered with FcγRIII (CD16) chimeric molecules to be redirected against malignant cells by monoclonal antibodies (mAbs). These cells have been proven to mediate granule-dependent cellular cytotoxicity, but it is not clear whether they can also kill malignant cells by a granule-independent mechanism of cell cytotoxicity. METHODS: We engineered a CD16A-CAR equipped with the extracellular CD16A, the hinge spacer and the transmembrane region of CD8, and the ζ-chain of the T-cell receptor/CD3 complex in tandem with the CD28 co-stimulatory signal transducer module. The CD16A-CAR was expressed and functionally tested in the MD45 cell line, a murine T-cell hybridoma with a defective granular exocytosis pathway but capable of killing target cells by a Fas ligand-mediated lysis. RESULTS: Our results indicate that in vitro cross-linking of CD16A-CAR on MD45 cells by the Fc fragment of mAb opsonized tumor cells induced interleukin-2 release and granule-independent cellular cytotoxicity. CONCLUSIONS: We conclude that strategies aimed to implement the therapeutic functions of mAbs used in the clinic with T-dependent immune responses driven by engineered T cells expressing FcγR-CAR can boost the antitumor efficacy of mAbs used in the clinic.
BACKGROUND AIMS: Chimeric antigen receptors (CARs) designed for adoptive immunotherapy need to achieve two functions: antigen recognition and triggering of the lytic machinery of reprogrammed effector cells. Cytotoxic T cells have been engineered with FcγRIII (CD16) chimeric molecules to be redirected against malignant cells by monoclonal antibodies (mAbs). These cells have been proven to mediate granule-dependent cellular cytotoxicity, but it is not clear whether they can also kill malignant cells by a granule-independent mechanism of cell cytotoxicity. METHODS: We engineered a CD16A-CAR equipped with the extracellular CD16A, the hinge spacer and the transmembrane region of CD8, and the ζ-chain of the T-cell receptor/CD3 complex in tandem with the CD28 co-stimulatory signal transducer module. The CD16A-CAR was expressed and functionally tested in the MD45 cell line, a murine T-cell hybridoma with a defective granular exocytosis pathway but capable of killing target cells by a Fas ligand-mediated lysis. RESULTS: Our results indicate that in vitro cross-linking of CD16A-CAR on MD45 cells by the Fc fragment of mAb opsonized tumor cells induced interleukin-2 release and granule-independent cellular cytotoxicity. CONCLUSIONS: We conclude that strategies aimed to implement the therapeutic functions of mAbs used in the clinic with T-dependent immune responses driven by engineered T cells expressing FcγR-CAR can boost the antitumor efficacy of mAbs used in the clinic.
Authors: Giuseppe Sconocchia; Giulia Lanzilli; Valeriana Cesarini; Domenico A Silvestris; Katayoun Rezvani; Roberto Arriga; Sara Caratelli; Ken Chen; Jinzhuang Dou; Carlo Cenciarelli; Gabriele Toietta; Silvia Baldari; Tommaso Sconocchia; Francesca De Paolis; Anna Aureli; Giandomenica Iezzi; Maria Irno Consalvo; Francesco Buccisano; Maria I Del Principe; Luca Maurillo; Adriano Venditti; Alessio Ottaviani; Giulio C Spagnoli Journal: Life Sci Alliance Date: 2022-10-14
Authors: Roberto Arriga; Sara Caratelli; Giulia Lanzilli; Alessio Ottaviani; Carlo Cenciarelli; Tommaso Sconocchia; Giulio C Spagnoli; Giandomenica Iezzi; Mario Roselli; Davide Lauro; Andrea Coppola; Gianpietro Dotti; Soldano Ferrone; Giuseppe Sconocchia Journal: Int J Cancer Date: 2019-08-30 Impact factor: 7.396
Authors: Sara Caratelli; Roberto Arriga; Tommaso Sconocchia; Alessio Ottaviani; Giulia Lanzilli; Donatella Pastore; Carlo Cenciarelli; Adriano Venditti; Maria Ilaria Del Principe; Davide Lauro; Elisa Landoni; Hongwei Du; Barbara Savoldo; Soldano Ferrone; Gianpietro Dotti; Giuseppe Sconocchia Journal: Int J Cancer Date: 2019-10-12 Impact factor: 7.396
Authors: Sara Caratelli; Tommaso Sconocchia; Roberto Arriga; Andrea Coppola; Giulia Lanzilli; Davide Lauro; Adriano Venditti; Maria Ilaria Del Principe; Francesco Buccisano; Luca Maurillo; Soldano Ferrone; Giuseppe Sconocchia Journal: Front Immunol Date: 2017-04-27 Impact factor: 7.561