title: Redirecting Specificity of T cells Using the Sleeping Beauty System to Express Chimeric Antigen Receptors by Mix-and-Matching of VL and VH Domains Targeting CD123+ Tumors.
abstract: Adoptive immunotherapy infusing T cells with engineered specificity for CD19 expressed on B- cell malignancies is generating enthusiasm to extend this approach to other hematological malignancies, such as acute myelogenous leukemia (AML). CD123, or interleukin 3 receptor alpha, is overexpressed on most AML and some lymphoid malignancies, such as acute lymphocytic leukemia (ALL), and has been an effective target for T cells expressing chimeric antigen receptors (CARs). The prototypical CAR encodes a VH and VL from one monoclonal antibody (mAb), coupled to a transmembrane domain and one or more cytoplasmic signaling domains. Previous studies showed that treatment of an experimental AML model with CD123-specific CAR T cells was therapeutic, but at the cost of impaired myelopoiesis, highlighting the need for systems to define the antigen threshold for CAR recognition. Here, we show that CARs can be engineered using VH and VL chains derived from different CD123-specific mAbs to generate a panel of CAR+ T cells. While all CARs exhibited specificity to CD123, one VH and VL combination had reduced lysis of normal hematopoietic stem cells. This CAR's in vivo anti-tumor activity was similar whether signaling occurred via chimeric CD28 or CD137, prolonging survival in both AML and ALL models. Co-expression of inducible caspase 9 eliminated CAR+ T cells. These data help support the use of CD123-specific CARs for treatment of CD123+ hematologic malignancies.
authors.affiliation: Division of Pediatrics, The University of Texas MD Anderson Cancer Center, Houston, Texas, United States of America.|The University of Texas Graduate School of Biomedical Sciences at Houston, Houston, Texas, United States of America.
authors.forename: Radhika
authors.identifier: http://orcid.org/0000-0002-4279-4115
authors.initials: R
authors.lastname: Thokala
authors.affiliation: Division of Pediatrics, The University of Texas MD Anderson Cancer Center, Houston, Texas, United States of America.
authors.forename: Simon
authors.initials: S
authors.lastname: Olivares
authors.affiliation: Division of Pediatrics, The University of Texas MD Anderson Cancer Center, Houston, Texas, United States of America.
authors.forename: Tiejuan
authors.initials: T
authors.lastname: Mi
authors.affiliation: Division of Pediatrics, The University of Texas MD Anderson Cancer Center, Houston, Texas, United States of America.
authors.forename: Sourindra
authors.initials: S
authors.lastname: Maiti
authors.affiliation: Division of Pediatrics, The University of Texas MD Anderson Cancer Center, Houston, Texas, United States of America.|Surgery Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, United States of America.
authors.forename: Drew
authors.initials: D
authors.lastname: Deniger
authors.affiliation: Intrexon Corporation, Germantown, Maryland, United States of America.
authors.forename: Helen
authors.initials: H
authors.lastname: Huls
authors.affiliation: Division of Pediatrics, The University of Texas MD Anderson Cancer Center, Houston, Texas, United States of America.
authors.forename: Hiroki
authors.initials: H
authors.lastname: Torikai
authors.affiliation: Division of Pediatrics, The University of Texas MD Anderson Cancer Center, Houston, Texas, United States of America.
authors.forename: Harjeet
authors.initials: H
authors.lastname: Singh
authors.affiliation: Stem Cell Transplantation and Cellular Therapy, The University of Texas MD Anderson Cancer Center, Houston, Texas, United States of America.
authors.forename: Richard E
authors.initials: RE
authors.lastname: Champlin
authors.affiliation: Division of Pediatrics, The University of Texas MD Anderson Cancer Center, Houston, Texas, United States of America.
authors.forename: Tamara
authors.initials: T
authors.lastname: Laskowski
authors.affiliation: Division of Pediatrics, The University of Texas MD Anderson Cancer Center, Houston, Texas, United States of America.
authors.forename: George
authors.initials: G
authors.lastname: McNamara
authors.affiliation: Division of Pediatrics, The University of Texas MD Anderson Cancer Center, Houston, Texas, United States of America.|Ziopharm Oncology Inc., Boston, Massachusetts, United States of America.
authors.forename: Laurence J N
authors.initials: LJ
authors.lastname: Cooper
mesh_terms: D000818:Animals; D001402:B-Lymphocytes / Q000276:immunology / Q000473:pathology; D018106:CD28 Antigens / Q000235:genetics / Q000276:immunology; D053453:Caspase 9 / Q000235:genetics / Q000276:immunology; D003602:Cytotoxicity, Immunologic; D004195:Disease Models, Animal; D015870:Gene Expression; D005818:Genetic Engineering / Q000379:methods*; D006412:Hematopoietic Stem Cells / Q000276:immunology / Q000473:pathology; D006801:Humans; D016219:Immunotherapy, Adoptive / Q000379:methods*; D053650:Interleukin-3 Receptor alpha Subunit / Q000235:genetics / Q000276:immunology*; D015470:Leukemia, Myeloid, Acute / Q000235:genetics / Q000276:immunology / Q000473:pathology / Q000628:therapy*; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D058990:Molecular Targeted Therapy; D010957:Plasmids; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000235:genetics / Q000276:immunology / Q000473:pathology / Q000628:therapy*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology*; D061905:Single-Domain Antibodies / Q000235:genetics; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology / Q000637:transplantation*; D014162:Transfection; D053261:Tumor Necrosis Factor Receptor Superfamily, Member 9 / Q000235:genetics / Q000276:immunology
chemical_list: D018106:CD28 Antigens; D053650:Interleukin-3 Receptor alpha Subunit; D011993:Recombinant Fusion Proteins; D061905:Single-Domain Antibodies; D053261:Tumor Necrosis Factor Receptor Superfamily, Member 9; C506433:CASP9 protein, human; D053453:Caspase 9