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Development and Validation of a Good Manufacturing Process for IL-4-Driven Expansion of Chimeric Cytokine Receptor-Expressing CAR T-Cells
Adoptive cancer immunotherapy using chimeric antigen receptor (CAR) engineered T-cells holds great promise, although several obstacles hinder the efficient generation of cell products under good manufacturing practice (GMP). Patients are often immune compromised, rendering it challenging to produce...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8307141/ https://www.ncbi.nlm.nih.gov/pubmed/34359966 http://dx.doi.org/10.3390/cells10071797 |
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author | van Schalkwyk, May C. I. van der Stegen, Sjoukje J. C. Bosshard-Carter, Leticia Graves, Helen Papa, Sophie Parente-Pereira, Ana C. Farzaneh, Farzin Fisher, Christopher D. Hope, Andrew Adami, Antonella Maher, John |
author_facet | van Schalkwyk, May C. I. van der Stegen, Sjoukje J. C. Bosshard-Carter, Leticia Graves, Helen Papa, Sophie Parente-Pereira, Ana C. Farzaneh, Farzin Fisher, Christopher D. Hope, Andrew Adami, Antonella Maher, John |
author_sort | van Schalkwyk, May C. I. |
collection | PubMed |
description | Adoptive cancer immunotherapy using chimeric antigen receptor (CAR) engineered T-cells holds great promise, although several obstacles hinder the efficient generation of cell products under good manufacturing practice (GMP). Patients are often immune compromised, rendering it challenging to produce sufficient numbers of gene-modified cells. Manufacturing protocols are labour intensive and frequently involve one or more open processing steps, leading to increased risk of contamination. We set out to develop a simplified process to generate autologous gamma retrovirus-transduced T-cells for clinical evaluation in patients with head and neck cancer. T-cells were engineered to co-express a panErbB-specific CAR (T1E28z) and a chimeric cytokine receptor (4αβ) that permits their selective expansion in response to interleukin (IL)-4. Using peripheral blood as starting material, sterile culture procedures were conducted in gas-permeable bags under static conditions. Pre-aliquoted medium and cytokines, bespoke connector devices and sterile welding/sealing were used to maximise the use of closed manufacturing steps. Reproducible IL-4-dependent expansion and enrichment of CAR-engineered T-cells under GMP was achieved, both from patients and healthy donors. We also describe the development and approach taken to validate a panel of monitoring and critical release assays, which provide objective data on cell product quality. |
format | Online Article Text |
id | pubmed-8307141 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-83071412021-07-25 Development and Validation of a Good Manufacturing Process for IL-4-Driven Expansion of Chimeric Cytokine Receptor-Expressing CAR T-Cells van Schalkwyk, May C. I. van der Stegen, Sjoukje J. C. Bosshard-Carter, Leticia Graves, Helen Papa, Sophie Parente-Pereira, Ana C. Farzaneh, Farzin Fisher, Christopher D. Hope, Andrew Adami, Antonella Maher, John Cells Article Adoptive cancer immunotherapy using chimeric antigen receptor (CAR) engineered T-cells holds great promise, although several obstacles hinder the efficient generation of cell products under good manufacturing practice (GMP). Patients are often immune compromised, rendering it challenging to produce sufficient numbers of gene-modified cells. Manufacturing protocols are labour intensive and frequently involve one or more open processing steps, leading to increased risk of contamination. We set out to develop a simplified process to generate autologous gamma retrovirus-transduced T-cells for clinical evaluation in patients with head and neck cancer. T-cells were engineered to co-express a panErbB-specific CAR (T1E28z) and a chimeric cytokine receptor (4αβ) that permits their selective expansion in response to interleukin (IL)-4. Using peripheral blood as starting material, sterile culture procedures were conducted in gas-permeable bags under static conditions. Pre-aliquoted medium and cytokines, bespoke connector devices and sterile welding/sealing were used to maximise the use of closed manufacturing steps. Reproducible IL-4-dependent expansion and enrichment of CAR-engineered T-cells under GMP was achieved, both from patients and healthy donors. We also describe the development and approach taken to validate a panel of monitoring and critical release assays, which provide objective data on cell product quality. MDPI 2021-07-15 /pmc/articles/PMC8307141/ /pubmed/34359966 http://dx.doi.org/10.3390/cells10071797 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article van Schalkwyk, May C. I. van der Stegen, Sjoukje J. C. Bosshard-Carter, Leticia Graves, Helen Papa, Sophie Parente-Pereira, Ana C. Farzaneh, Farzin Fisher, Christopher D. Hope, Andrew Adami, Antonella Maher, John Development and Validation of a Good Manufacturing Process for IL-4-Driven Expansion of Chimeric Cytokine Receptor-Expressing CAR T-Cells |
title | Development and Validation of a Good Manufacturing Process for IL-4-Driven Expansion of Chimeric Cytokine Receptor-Expressing CAR T-Cells |
title_full | Development and Validation of a Good Manufacturing Process for IL-4-Driven Expansion of Chimeric Cytokine Receptor-Expressing CAR T-Cells |
title_fullStr | Development and Validation of a Good Manufacturing Process for IL-4-Driven Expansion of Chimeric Cytokine Receptor-Expressing CAR T-Cells |
title_full_unstemmed | Development and Validation of a Good Manufacturing Process for IL-4-Driven Expansion of Chimeric Cytokine Receptor-Expressing CAR T-Cells |
title_short | Development and Validation of a Good Manufacturing Process for IL-4-Driven Expansion of Chimeric Cytokine Receptor-Expressing CAR T-Cells |
title_sort | development and validation of a good manufacturing process for il-4-driven expansion of chimeric cytokine receptor-expressing car t-cells |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8307141/ https://www.ncbi.nlm.nih.gov/pubmed/34359966 http://dx.doi.org/10.3390/cells10071797 |
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