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.

Magnani, C., Turazzi, N., Benedicenti, F., Calabria, A., Tenderini, E., Tettamanti, S., et al. (2016). Immunotherapy of acute leukemia by chimeric antigen receptormodified lymphocytes using an improved Sleeping Beauty transposon platform. ONCOTARGET, 7(32), 51581-51597 [10.18632/oncotarget.9955].

Immunotherapy of acute leukemia by chimeric antigen receptormodified lymphocytes using an improved Sleeping Beauty transposon platform

Magnani, C;Turazzi, N;Calabria, A;Biondi, A
;
Biagi, E
2016

Abstract

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.
Articolo in rivista - Articolo scientifico
Acute lymphoblastic leukemia; Acute myelogenous leukemia; Chimeric antigen receptor; Cytokine-induced killer cells; Sleeping Beauty transposon;
Sleeping Beauty transposon; acute lymphoblastic leukemia; acute myelogenous leukemia; chimeric antigen receptor; cytokine-induced killer cells
English
2016
7
32
51581
51597
none
Magnani, C., Turazzi, N., Benedicenti, F., Calabria, A., Tenderini, E., Tettamanti, S., et al. (2016). Immunotherapy of acute leukemia by chimeric antigen receptormodified lymphocytes using an improved Sleeping Beauty transposon platform. ONCOTARGET, 7(32), 51581-51597 [10.18632/oncotarget.9955].
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/10281/122933
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