Immunotherapy of acute leukemia by chimeric antigen receptor-modified lymphocytes using an improved Sleeping Beauty transposon platform

Chimeric antigen receptor (CAR)-modified T-cell adoptive immunotherapy is a remarkable therapeutic option proven effective in the treatment of hematological malignancies. In order to optimize cell manufacturing, we sought to develop a novel clinical-grade protocol to obtain CAR-modified cytokine-ind...

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Bibliographic Details
Published in:Oncotarget 2016-08, Vol.7 (32), p.51581-51597
Main Authors: Magnani, Chiara F, Turazzi, Nice, Benedicenti, Fabrizio, Calabria, Andrea, Tenderini, Erika, Tettamanti, Sarah, Giordano Attianese, Greta M P, Cooper, Laurence J N, Aiuti, Alessandro, Montini, Eugenio, Biondi, Andrea, Biagi, Ettore
Format: Article
Language:English
Subjects:
Acute Disease
Adolescent
Animals
Antigens, CD19 - genetics
Antigens, CD19 - metabolism
Cell Line, Tumor
Child
Child, Preschool
Combined Modality Therapy
Female
Genetic Therapy - methods
Humans
Immunotherapy, Adoptive - methods
Infant
Interleukin-3 Receptor alpha Subunit - genetics
Interleukin-3 Receptor alpha Subunit - metabolism
Leukemia - immunology
Leukemia - pathology
Leukemia - therapy
Mice
Mice, Inbred NOD
Mice, SCID
Mice, Transgenic
Receptors, Antigen, T-Cell - genetics
Receptors, Antigen, T-Cell - immunology
Receptors, Antigen, T-Cell - metabolism
Research Paper
T-Lymphocytes - metabolism
T-Lymphocytes - transplantation
Transposases - genetics
Transposases - metabolism
Xenograft Model Antitumor Assays
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Summary:Chimeric antigen receptor (CAR)-modified T-cell adoptive immunotherapy is a remarkable therapeutic option proven effective in the treatment of hematological malignancies. In order to optimize cell manufacturing, we sought to develop a novel clinical-grade protocol to obtain CAR-modified cytokine-induced killer cells (CIKs) using the Sleeping Beauty (SB) transposon system. Administration of irradiated PBMCs overcame cell death of stimulating cells induced by non-viral transfection, enabling robust gene transfer together with efficient T-cell expansion. Upon single stimulation, we reached an average of 60% expression of CD123- and CD19- specific 3rd generation CARs (CD28/OX40/TCRzeta). Furthermore, modified cells displayed persistence of cell subsets with memory phenotype, specific and effective lytic activity against leukemic cell lines and primary blasts, cytokine secretion, and proliferation. Adoptive transfer of CD123.CAR or CD19.CAR lymphocytes led to a significant anti-tumor response against acute myelogenous leukemia (AML) and acute lymphoblastic leukemia (ALL) disseminated diseases in NSG mice. Notably, we found no evidence of integration enrichment near cancer genes and transposase expression at the end of the differentiation. Taken all together, our findings describe a novel donor-derived non-viral CAR approach that may widen the repertoire of available methods for T cell-based immunotherapy.
ISSN:1949-2553
1949-2553
DOI:10.18632/oncotarget.9955