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...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Nature Publishing Group
2016
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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 |
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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 |
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