title: A novel anti-GD2/4-1BB chimeric antigen receptor triggers neuroblastoma cell killing.
abstract: Chimeric antigen receptor (CAR)-expressing T cells are a promising therapeutic option for patients with cancer. We developed a new CAR directed against the disialoganglioside GD2, a surface molecule expressed in neuroblastoma and in other neuroectoderm-derived neoplasms. The anti-GD2 single-chain variable fragment (scFv) derived from a murine antibody of IgM class was linked, via a human CD8α hinge-transmembrane domain, to the signaling domains of the costimulatory molecules 4-1BB (CD137) and CD3-ζ. The receptor was expressed in T lymphocytes by retroviral transduction and anti-tumor activities were assessed by targeting GD2-positive neuroblastoma cells using in vitro cytotoxicity assays and a xenograft model. Transduced T cells expressed high levels of anti-GD2 CAR and exerted a robust and specific anti-tumor activity in 4- and 48-hour cultures with neuroblastoma cells. Cytotoxicity was associated with the release of pro-apoptotic molecules such as TRAIL and IFN-γ. These results were confirmed in a xenograft model, where anti-GD2 CAR T cells infiltrating tumors and persisting into blood circulation induced massive apoptosis of neuroblastoma cells and completely abrogated tumor growth. This anti-GD2 CAR represents a powerful new tool to redirect T cells against GD2. The preclinical results of this study warrant clinical testing of this approach in neuroblastoma and other GD2-positive malignancies.
authors: Department of Medical and Surgical Sciences for Children & Adults, Division of Oncology, University-Hospital of Modena and Reggio Emilia, Modena, Italy.
authors: Malvina
authors: M
authors: Prapa
authors: Department of Pathology and Diagnostics, University of Verona, Verona, Italy.
authors: Sara
authors: S
authors: Caldrer
authors: Department of Medical and Surgical Sciences for Children & Adults, Division of Oncology, University-Hospital of Modena and Reggio Emilia, Modena, Italy.
authors: Carlotta
authors: C
authors: Spano
authors: International Centre for Genetic Engineering and Biotechnology, Trieste, Italy.
authors: Marco
authors: M
authors: Bestagno
authors: Department of Medical and Surgical Sciences for Children & Adults, Division of Oncology, University-Hospital of Modena and Reggio Emilia, Modena, Italy.
authors: Giulia
authors: G
authors: Golinelli
authors: Department of Medical and Surgical Sciences for Children & Adults, Division of Oncology, University-Hospital of Modena and Reggio Emilia, Modena, Italy.
authors: Giulia
authors: G
authors: Grisendi
authors: Department of Medical and Surgical Sciences for Children & Adults, Division of Oncology, University-Hospital of Modena and Reggio Emilia, Modena, Italy.
authors: Tiziana
authors: T
authors: Petrachi
authors: Istituto Oncologico Veneto, Padova, Italy.
authors: Pierfranco
authors: P
authors: Conte
authors: Departments of Pediatrics and Medicine, Division of Hematology/Oncology/BMT, Nationwide Children's Hospital, The Ohio State University College of Medicine, Columbus, Ohio, USA.
authors: Edwin M
authors: EM
authors: Horwitz
authors: Department of Pediatrics, National University of Singapore, Singapore.
authors: Dario
authors: D
authors: Campana
authors: Department of Medical and Surgical Sciences for Children & Adults, Division of Oncology, University-Hospital of Modena and Reggio Emilia, Modena, Italy.
authors: Paolo
authors: P
authors: Paolucci
authors: Department of Medical and Surgical Sciences for Children & Adults, Division of Oncology, University-Hospital of Modena and Reggio Emilia, Modena, Italy.
authors: Massimo
authors: M
authors: Dominici
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000276:immunology; D045744:Cell Line, Tumor; D003602:Cytotoxicity, Immunologic / Q000276:immunology; D005434:Flow Cytometry; D005732:Gangliosides / Q000276:immunology*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D016219:Immunotherapy, Adoptive / Q000379:methods; D007371:Interferon-gamma / Q000276:immunology / Q000378:metabolism; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D008856:Microscopy, Fluorescence; D009447:Neuroblastoma / Q000276:immunology* / Q000473:pathology / Q000628:therapy; D011948:Receptors, Antigen, T-Cell / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D057127:Single-Chain Antibodies / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D013601:T-Lymphocytes / Q000276:immunology* / Q000378:metabolism / Q000637:transplantation; D053221:TNF-Related Apoptosis-Inducing Ligand / Q000276:immunology / Q000378:metabolism; D053261:Tumor Necrosis Factor Receptor Superfamily, Member 9 / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D023041:Xenograft Model Antitumor Assays
chemical_list: D005732:Gangliosides; D011948:Receptors, Antigen, T-Cell; D057127:Single-Chain Antibodies; D053221:TNF-Related Apoptosis-Inducing Ligand; C505653:TNFSF10 protein, human; D053261:Tumor Necrosis Factor Receptor Superfamily, Member 9; D049452:Green Fluorescent Proteins; C019403:ganglioside, GD2; D007371:Interferon-gamma
keywords: GD2; T lymphocytes; anti-GD2 IgM-derived; chimeric antigen receptor; neuroblastoma