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-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.
Tettamanti Research Center, Department of Pediatrics, University of Milano-Bicocca, San Gerardo Hospital/Fondazione MBBM, Monza, Italy.
Chiara F
CF
Magnani
Tettamanti Research Center, Department of Pediatrics, University of Milano-Bicocca, San Gerardo Hospital/Fondazione MBBM, Monza, Italy.
Nice
N
Turazzi
San Raffaele Telethon Institute for Gene Therapy (HSR-TIGET), Milan, Italy.
Fabrizio
F
Benedicenti
San Raffaele Telethon Institute for Gene Therapy (HSR-TIGET), Milan, Italy.
Andrea
A
Calabria
San Raffaele Telethon Institute for Gene Therapy (HSR-TIGET), Milan, Italy.
Erika
E
Tenderini
Tettamanti Research Center, Department of Pediatrics, University of Milano-Bicocca, San Gerardo Hospital/Fondazione MBBM, Monza, Italy.
Sarah
S
Tettamanti
Tettamanti Research Center, Department of Pediatrics, University of Milano-Bicocca, San Gerardo Hospital/Fondazione MBBM, Monza, Italy.|University of Texas MD Anderson Cancer Center, Houston, Texas, USA.
Greta M P
GM
Giordano Attianese
University of Texas MD Anderson Cancer Center, Houston, Texas, USA.
Laurence J N
LJ
Cooper
San Raffaele Telethon Institute for Gene Therapy (HSR-TIGET), Milan, Italy.
Alessandro
A
Aiuti
San Raffaele Telethon Institute for Gene Therapy (HSR-TIGET), Milan, Italy.
Eugenio
E
Montini
Tettamanti Research Center, Department of Pediatrics, University of Milano-Bicocca, San Gerardo Hospital/Fondazione MBBM, Monza, Italy.
Andrea
A
Biondi
Tettamanti Research Center, Department of Pediatrics, University of Milano-Bicocca, San Gerardo Hospital/Fondazione MBBM, Monza, Italy.
Ettore
E
Biagi
D000208:Acute Disease; D000293:Adolescent; D000818:Animals; D018941:Antigens, CD19 / Q000235:genetics / Q000378:metabolism; D045744:Cell Line, Tumor; D002648:Child; D002675:Child, Preschool; D003131:Combined Modality Therapy; D005260:Female; D015316:Genetic Therapy / Q000379:methods*; D006801:Humans; D016219:Immunotherapy, Adoptive / Q000379:methods*; D007223:Infant; D053650:Interleukin-3 Receptor alpha Subunit / Q000235:genetics / Q000378:metabolism; D007938:Leukemia / Q000276:immunology / Q000473:pathology* / Q000628:therapy*; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D008822:Mice, Transgenic; D011948:Receptors, Antigen, T-Cell / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D013601:T-Lymphocytes / Q000378:metabolism / Q000637:transplantation*; D019895:Transposases / Q000235:genetics* / Q000378:metabolism; D023041:Xenograft Model Antitumor Assays
D018941:Antigens, CD19; C000604811:CD19 molecule, human; C115022:IL3RA protein, human; D053650:Interleukin-3 Receptor alpha Subunit; D011948:Receptors, Antigen, T-Cell; D019895:Transposases; C494471:sleeping beauty transposase, human
Sleeping Beauty transposon; acute lymphoblastic leukemia; acute myelogenous leukemia; chimeric antigen receptor; cytokine-induced killer cells