Preclinical validation: LV/IL-12 transduction of patient leukemia cells for immunotherapy of AML

Interleukin-12 (IL-12) is a potent cytokine that may be harnessed to treat cancer. To date, nearly 100 IL-12-based clinical trials have been initiated worldwide. Yet systemic administration of IL-12 is toxic. Different strategies are being developed to reduce such toxicities by restricting IL-12 dis...

Descripción completa

Detalles Bibliográficos
Autores principales: Huang, Ju, Liu, Yuanfeng, Au, Bryan C, Barber, Dwayne L, Arruda, Andrea, Schambach, Axel, Rothe, Michael, Minden, Mark D, Paige, Christopher J, Medin, Jeffrey A
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5142463/
https://www.ncbi.nlm.nih.gov/pubmed/27933304
http://dx.doi.org/10.1038/mtm.2016.74
_version_ 1782472777774661632
author Huang, Ju
Liu, Yuanfeng
Au, Bryan C
Barber, Dwayne L
Arruda, Andrea
Schambach, Axel
Rothe, Michael
Minden, Mark D
Paige, Christopher J
Medin, Jeffrey A
author_facet Huang, Ju
Liu, Yuanfeng
Au, Bryan C
Barber, Dwayne L
Arruda, Andrea
Schambach, Axel
Rothe, Michael
Minden, Mark D
Paige, Christopher J
Medin, Jeffrey A
author_sort Huang, Ju
collection PubMed
description Interleukin-12 (IL-12) is a potent cytokine that may be harnessed to treat cancer. To date, nearly 100 IL-12-based clinical trials have been initiated worldwide. Yet systemic administration of IL-12 is toxic. Different strategies are being developed to reduce such toxicities by restricting IL-12 distribution. Our previous studies employed lentivector-mediated expression of murine IL-12 in tumor cells and demonstrated effective protection in both mouse leukemia and solid tumor challenge models. In this study, we carried out preclinical validation studies using a novel lentivector to engineer expression of human IL-12 in acute myeloid leukemia blast cells isolated from 21 patients. Acute myeloid leukemia cells were transduced with a bicistronic lentivector that encodes the human IL-12 cDNA as a fusion, as well as a LNGFR (ΔLNGFR)/mutant thymidylate kinase cassette as a marking and cell-fate control element. A range of 20–70% functional transduction efficiencies was achieved. Transduced acute myeloid leukemia cells produced bioactive IL-12 protein and displayed dose-dependent sensitivity to the prodrug 3′-azido-3′-deoxythymidine. In vitro immortalization assays using transduced mouse hematopoietic stem cells demonstrated minimal genotoxic risk from our IL-12 vector. Scale-up transduction and cell processing was subsequently validated in a GMP facility to support our (now approved) Clinical Trial Application (CTA).
format Online
Article
Text
id pubmed-5142463
institution National Center for Biotechnology Information
language English
publishDate 2016
publisher Nature Publishing Group
record_format MEDLINE/PubMed
spelling pubmed-51424632016-12-08 Preclinical validation: LV/IL-12 transduction of patient leukemia cells for immunotherapy of AML Huang, Ju Liu, Yuanfeng Au, Bryan C Barber, Dwayne L Arruda, Andrea Schambach, Axel Rothe, Michael Minden, Mark D Paige, Christopher J Medin, Jeffrey A Mol Ther Methods Clin Dev Article Interleukin-12 (IL-12) is a potent cytokine that may be harnessed to treat cancer. To date, nearly 100 IL-12-based clinical trials have been initiated worldwide. Yet systemic administration of IL-12 is toxic. Different strategies are being developed to reduce such toxicities by restricting IL-12 distribution. Our previous studies employed lentivector-mediated expression of murine IL-12 in tumor cells and demonstrated effective protection in both mouse leukemia and solid tumor challenge models. In this study, we carried out preclinical validation studies using a novel lentivector to engineer expression of human IL-12 in acute myeloid leukemia blast cells isolated from 21 patients. Acute myeloid leukemia cells were transduced with a bicistronic lentivector that encodes the human IL-12 cDNA as a fusion, as well as a LNGFR (ΔLNGFR)/mutant thymidylate kinase cassette as a marking and cell-fate control element. A range of 20–70% functional transduction efficiencies was achieved. Transduced acute myeloid leukemia cells produced bioactive IL-12 protein and displayed dose-dependent sensitivity to the prodrug 3′-azido-3′-deoxythymidine. In vitro immortalization assays using transduced mouse hematopoietic stem cells demonstrated minimal genotoxic risk from our IL-12 vector. Scale-up transduction and cell processing was subsequently validated in a GMP facility to support our (now approved) Clinical Trial Application (CTA). Nature Publishing Group 2016-12-07 /pmc/articles/PMC5142463/ /pubmed/27933304 http://dx.doi.org/10.1038/mtm.2016.74 Text en Copyright © 2016 Official journal of the American Society of Gene & Cell Therapy http://creativecommons.org/licenses/by-nc-nd/4.0/ This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-nd/4.0/
spellingShingle Article
Huang, Ju
Liu, Yuanfeng
Au, Bryan C
Barber, Dwayne L
Arruda, Andrea
Schambach, Axel
Rothe, Michael
Minden, Mark D
Paige, Christopher J
Medin, Jeffrey A
Preclinical validation: LV/IL-12 transduction of patient leukemia cells for immunotherapy of AML
title Preclinical validation: LV/IL-12 transduction of patient leukemia cells for immunotherapy of AML
title_full Preclinical validation: LV/IL-12 transduction of patient leukemia cells for immunotherapy of AML
title_fullStr Preclinical validation: LV/IL-12 transduction of patient leukemia cells for immunotherapy of AML
title_full_unstemmed Preclinical validation: LV/IL-12 transduction of patient leukemia cells for immunotherapy of AML
title_short Preclinical validation: LV/IL-12 transduction of patient leukemia cells for immunotherapy of AML
title_sort preclinical validation: lv/il-12 transduction of patient leukemia cells for immunotherapy of aml
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5142463/
https://www.ncbi.nlm.nih.gov/pubmed/27933304
http://dx.doi.org/10.1038/mtm.2016.74
work_keys_str_mv AT huangju preclinicalvalidationlvil12transductionofpatientleukemiacellsforimmunotherapyofaml
AT liuyuanfeng preclinicalvalidationlvil12transductionofpatientleukemiacellsforimmunotherapyofaml
AT aubryanc preclinicalvalidationlvil12transductionofpatientleukemiacellsforimmunotherapyofaml
AT barberdwaynel preclinicalvalidationlvil12transductionofpatientleukemiacellsforimmunotherapyofaml
AT arrudaandrea preclinicalvalidationlvil12transductionofpatientleukemiacellsforimmunotherapyofaml
AT schambachaxel preclinicalvalidationlvil12transductionofpatientleukemiacellsforimmunotherapyofaml
AT rothemichael preclinicalvalidationlvil12transductionofpatientleukemiacellsforimmunotherapyofaml
AT mindenmarkd preclinicalvalidationlvil12transductionofpatientleukemiacellsforimmunotherapyofaml
AT paigechristopherj preclinicalvalidationlvil12transductionofpatientleukemiacellsforimmunotherapyofaml
AT medinjeffreya preclinicalvalidationlvil12transductionofpatientleukemiacellsforimmunotherapyofaml