TY - JOUR
T1 - Immunotherapy of acute leukemia by chimeric antigen receptormodified lymphocytes using an improved Sleeping Beauty transposon platform
AU - Magnani, Chiara F.
AU - Turazzi, Nice
AU - Benedicenti, Fabrizio
AU - Calabria, Andrea
AU - Tenderini, Erika
AU - Tettamanti, Sarah
AU - Giordano Attianese, Greta M P
AU - Cooper, Laurence J N
AU - Aiuti, Alessandro
AU - Montini, Eugenio
AU - Biondi, Andrea
AU - Biagi, Ettore
PY - 2016/8/1
Y1 - 2016/8/1
N2 - 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.
AB - 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.
KW - Acute lymphoblastic leukemia
KW - Acute myelogenous leukemia
KW - Chimeric antigen receptor
KW - Cytokine-induced killer cells
KW - Sleeping Beauty transposon
UR - http://www.scopus.com/inward/record.url?scp=84982291277&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84982291277&partnerID=8YFLogxK
U2 - 10.18632/oncotarget.9955
DO - 10.18632/oncotarget.9955
M3 - Article
AN - SCOPUS:84982291277
VL - 7
SP - 51581
EP - 51597
JO - Oncotarget
JF - Oncotarget
SN - 1949-2553
IS - 32
ER -