title: Sleeping Beauty Transposition of Chimeric Antigen Receptors Targeting Receptor Tyrosine Kinase-Like Orphan Receptor-1 (ROR1) into Diverse Memory T-Cell Populations.
abstract: T cells modified with chimeric antigen receptors (CARs) targeting CD19 demonstrated clinical activity against some B-cell malignancies. However, this is often accompanied by a loss of normal CD19+ B cells and humoral immunity. Receptor tyrosine kinase-like orphan receptor-1 (ROR1) is expressed on sub-populations of B-cell malignancies and solid tumors, but not by healthy B cells or normal post-partum tissues. Thus, adoptive transfer of T cells specific for ROR1 has potential to eliminate tumor cells and spare healthy tissues. To test this hypothesis, we developed CARs targeting ROR1 in order to generate T cells specific for malignant cells. Two Sleeping Beauty transposons were constructed with 2nd generation ROR1-specific CARs signaling through CD3ζ and either CD28 (designated ROR1RCD28) or CD137 (designated ROR1RCD137) and were introduced into T cells. We selected for T cells expressing CAR through co-culture with γ-irradiated activating and propagating cells (AaPC), which co-expressed ROR1 and co-stimulatory molecules. Numeric expansion over one month of co-culture on AaPC in presence of soluble interleukin (IL)-2 and IL-21 occurred and resulted in a diverse memory phenotype of CAR+ T cells as measured by non-enzymatic digital array (NanoString) and multi-panel flow cytometry. Such T cells produced interferon-γ and had specific cytotoxic activity against ROR1+ tumors. Moreover, such cells could eliminate ROR1+ tumor xenografts, especially T cells expressing ROR1RCD137. Clinical trials will investigate the ability of ROR1-specific CAR+ T cells to specifically eliminate tumor cells while maintaining normal B-cell repertoire.
authors.affiliation: Pediatrics, Children's Cancer Hospital, University of Texas MD Anderson Cancer Center, Houston, Texas, United States of America; University of Texas Graduate School of Biomedical Sciences at Houston, Houston, Texas, United States of America.
authors.forename: Drew C
authors.initials: DC
authors.lastname: Deniger
authors.affiliation: Medicine, Moores Cancer Center, University of California San Diego, San Diego, California, United States of America.
authors.forename: Jianqiang
authors.initials: J
authors.lastname: Yu
authors.affiliation: Pediatrics, Children's Cancer Hospital, University of Texas MD Anderson Cancer Center, Houston, Texas, United States of America.
authors.forename: M Helen
authors.initials: MH
authors.lastname: Huls
authors.affiliation: Pediatrics, Children's Cancer Hospital, University of Texas MD Anderson Cancer Center, Houston, Texas, United States of America.
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Figliola
authors.affiliation: Pediatrics, Children's Cancer Hospital, University of Texas MD Anderson Cancer Center, Houston, Texas, United States of America.
authors.forename: Tiejuan
authors.initials: T
authors.lastname: Mi
authors.affiliation: Pediatrics, Children's Cancer Hospital, University of Texas MD Anderson Cancer Center, Houston, Texas, United States of America.
authors.forename: Sourindra N
authors.initials: SN
authors.lastname: Maiti
authors.affiliation: Medicine, Moores Cancer Center, University of California San Diego, San Diego, California, United States of America.
authors.forename: George F
authors.initials: GF
authors.lastname: Widhopf
authors.affiliation: Pediatrics, Children's Cancer Hospital, University of Texas MD Anderson Cancer Center, Houston, Texas, United States of America; University of Texas Graduate School of Biomedical Sciences at Houston, Houston, Texas, United States of America.
authors.forename: Lenka V
authors.initials: LV
authors.lastname: Hurton
authors.affiliation: Pediatrics, Children's Cancer Hospital, University of Texas MD Anderson Cancer Center, Houston, Texas, United States of America; University of Texas Graduate School of Biomedical Sciences at Houston, Houston, Texas, United States of America.
authors.forename: Radhika
authors.initials: R
authors.lastname: Thokala
authors.affiliation: Pediatrics, Children's Cancer Hospital, University of Texas MD Anderson Cancer Center, Houston, Texas, United States of America.
authors.forename: Harjeet
authors.initials: H
authors.lastname: Singh
authors.affiliation: Pediatrics, Children's Cancer Hospital, University of Texas MD Anderson Cancer Center, Houston, Texas, United States of America.
authors.forename: Simon
authors.initials: S
authors.lastname: Olivares
authors.affiliation: Stem Cell Transplantation and Cellular Therapy, 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: Leukemia, University of Texas MD Anderson Cancer Center, Houston, Texas, United States of America.
authors.forename: William G
authors.initials: WG
authors.lastname: Wierda
authors.affiliation: Medicine, Moores Cancer Center, University of California San Diego, San Diego, California, United States of America.
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Kipps
authors.affiliation: Pediatrics, Children's Cancer Hospital, University of Texas MD Anderson Cancer Center, Houston, Texas, United States of America; University of Texas Graduate School of Biomedical Sciences at Houston, Houston, Texas, United States of America.
authors.forename: Laurence J N
authors.initials: LJ
authors.lastname: Cooper
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D018920:Coculture Techniques; D016207:Cytokines / Q000378:metabolism; D004251:DNA Transposable Elements / Q000235:genetics*; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007371:Interferon-gamma / Q000096:biosynthesis; D007938:Leukemia / Q000276:immunology; D051379:Mice; D010190:Pancreatic Neoplasms / Q000235:genetics / Q000276:immunology / Q000473:pathology; D010641:Phenotype; D057050:Receptor Tyrosine Kinase-like Orphan Receptors / Q000378:metabolism*; D011948:Receptors, Antigen, T-Cell / Q000235:genetics* / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000378:metabolism*; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology / Q000378:metabolism*; D014158:Transcription, Genetic
chemical_list: D016207:Cytokines; D004251:DNA Transposable Elements; D011948:Receptors, Antigen, T-Cell; D011993:Recombinant Fusion Proteins; D007371:Interferon-gamma; D057050:Receptor Tyrosine Kinase-like Orphan Receptors