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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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Autores principales: Magnani, Chiara F., Turazzi, Nice, Benedicenti, Fabrizio, Calabria, Andrea, Tenderini, Erika, Tettamanti, Sarah, Attianese, Greta M.P. Giordano, Cooper, Laurence J.N., Aiuti, Alessandro, Montini, Eugenio, Biondi, Andrea, Biagi, Ettore
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Impact Journals LLC 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5239498/
https://www.ncbi.nlm.nih.gov/pubmed/27323395
http://dx.doi.org/10.18632/oncotarget.9955
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author Magnani, Chiara F.
Turazzi, Nice
Benedicenti, Fabrizio
Calabria, Andrea
Tenderini, Erika
Tettamanti, Sarah
Attianese, Greta M.P. Giordano
Cooper, Laurence J.N.
Aiuti, Alessandro
Montini, Eugenio
Biondi, Andrea
Biagi, Ettore
author_facet Magnani, Chiara F.
Turazzi, Nice
Benedicenti, Fabrizio
Calabria, Andrea
Tenderini, Erika
Tettamanti, Sarah
Attianese, Greta M.P. Giordano
Cooper, Laurence J.N.
Aiuti, Alessandro
Montini, Eugenio
Biondi, Andrea
Biagi, Ettore
author_sort Magnani, Chiara F.
collection PubMed
description 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.
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spelling pubmed-52394982017-01-24 Immunotherapy of acute leukemia by chimeric antigen receptor-modified lymphocytes using an improved Sleeping Beauty transposon platform Magnani, Chiara F. Turazzi, Nice Benedicenti, Fabrizio Calabria, Andrea Tenderini, Erika Tettamanti, Sarah Attianese, Greta M.P. Giordano Cooper, Laurence J.N. Aiuti, Alessandro Montini, Eugenio Biondi, Andrea Biagi, Ettore Oncotarget Research Paper 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. Impact Journals LLC 2016-06-13 /pmc/articles/PMC5239498/ /pubmed/27323395 http://dx.doi.org/10.18632/oncotarget.9955 Text en Copyright: © 2016 Magnani et al. http://creativecommons.org/licenses/by/2.5/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Paper
Magnani, Chiara F.
Turazzi, Nice
Benedicenti, Fabrizio
Calabria, Andrea
Tenderini, Erika
Tettamanti, Sarah
Attianese, Greta M.P. Giordano
Cooper, Laurence J.N.
Aiuti, Alessandro
Montini, Eugenio
Biondi, Andrea
Biagi, Ettore
Immunotherapy of acute leukemia by chimeric antigen receptor-modified lymphocytes using an improved Sleeping Beauty transposon platform
title Immunotherapy of acute leukemia by chimeric antigen receptor-modified lymphocytes using an improved Sleeping Beauty transposon platform
title_full Immunotherapy of acute leukemia by chimeric antigen receptor-modified lymphocytes using an improved Sleeping Beauty transposon platform
title_fullStr Immunotherapy of acute leukemia by chimeric antigen receptor-modified lymphocytes using an improved Sleeping Beauty transposon platform
title_full_unstemmed Immunotherapy of acute leukemia by chimeric antigen receptor-modified lymphocytes using an improved Sleeping Beauty transposon platform
title_short Immunotherapy of acute leukemia by chimeric antigen receptor-modified lymphocytes using an improved Sleeping Beauty transposon platform
title_sort immunotherapy of acute leukemia by chimeric antigen receptor-modified lymphocytes using an improved sleeping beauty transposon platform
topic Research Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5239498/
https://www.ncbi.nlm.nih.gov/pubmed/27323395
http://dx.doi.org/10.18632/oncotarget.9955
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