title: Redirecting specificity of T-cell populations for CD19 using the Sleeping Beauty system.
abstract: Genetic modification of clinical-grade T cells is undertaken to augment function, including redirecting specificity for desired antigen. We and others have introduced a chimeric antigen receptor (CAR) to enable T cells to recognize lineage-specific tumor antigen, such as CD19, and early-phase human trials are currently assessing safety and feasibility. However, a significant barrier to next-generation clinical studies is developing a suitable CAR expression vector capable of genetically modifying a broad population of T cells. Transduction of T cells is relatively efficient but it requires specialized manufacture of expensive clinical grade recombinant virus. Electrotransfer of naked DNA plasmid offers a cost-effective alternative approach, but the inefficiency of transgene integration mandates ex vivo selection under cytocidal concentrations of drug to enforce expression of selection genes to achieve clinically meaningful numbers of CAR(+) T cells. We report a new approach to efficiently generating T cells with redirected specificity, introducing DNA plasmids from the Sleeping Beauty transposon/transposase system to directly express a CD19-specific CAR in memory and effector T cells without drug selection. When coupled with numerical expansion on CD19(+) artificial antigen-presenting cells, this gene transfer method results in rapid outgrowth of CD4(+) and CD8(+) T cells expressing CAR to redirect specificity for CD19(+) tumor cells.
authors.affiliation: Division of Pediatrics, University of Texas M. D. Anderson Cancer Center, Houston, TX 77030, USA.
authors.forename: Harjeet
authors.initials: H
authors.lastname: Singh
authors.forename: Pallavi R
authors.initials: PR
authors.lastname: Manuri
authors.forename: Simon
authors.initials: S
authors.lastname: Olivares
authors.forename: Navid
authors.initials: N
authors.lastname: Dara
authors.forename: Margaret J
authors.initials: MJ
authors.lastname: Dawson
authors.forename: Helen
authors.initials: H
authors.lastname: Huls
authors.forename: Perry B
authors.initials: PB
authors.lastname: Hackett
authors.forename: Donald B
authors.initials: DB
authors.lastname: Kohn
authors.forename: Elizabeth J
authors.initials: EJ
authors.lastname: Shpall
authors.forename: Richard E
authors.initials: RE
authors.lastname: Champlin
authors.forename: Laurence J N
authors.initials: LJ
authors.lastname: Cooper
mesh_terms: D018941:Antigens, CD19 / Q000276:immunology*; D000951:Antigens, Neoplasm / Q000276:immunology*; D002051:Burkitt Lymphoma / Q000276:immunology; D045744:Cell Line, Tumor / Q000276:immunology; D018920:Coculture Techniques; D003062:Codon / Q000276:immunology; D006412:Hematopoietic Stem Cells / Q000276:immunology; D006801:Humans; D020014:K562 Cells / Q000276:immunology; D017710:Lymphocyte Transfusion; D009369:Neoplasms / Q000276:immunology*; D033581:Stem Cell Transplantation; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology*; D014184:Transplantation, Homologous; D019895:Transposases / Q000276:immunology*
chemical_list: D018941:Antigens, CD19; D000951:Antigens, Neoplasm; D003062:Codon; D019895:Transposases; C494471:sleeping beauty transposase, human