Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: 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
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
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
_version_ 1783727979423072256
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
work_keys_str_mv AT vanschalkwykmayci developmentandvalidationofagoodmanufacturingprocessforil4drivenexpansionofchimericcytokinereceptorexpressingcartcells
AT vanderstegensjoukjejc developmentandvalidationofagoodmanufacturingprocessforil4drivenexpansionofchimericcytokinereceptorexpressingcartcells
AT bosshardcarterleticia developmentandvalidationofagoodmanufacturingprocessforil4drivenexpansionofchimericcytokinereceptorexpressingcartcells
AT graveshelen developmentandvalidationofagoodmanufacturingprocessforil4drivenexpansionofchimericcytokinereceptorexpressingcartcells
AT papasophie developmentandvalidationofagoodmanufacturingprocessforil4drivenexpansionofchimericcytokinereceptorexpressingcartcells
AT parentepereiraanac developmentandvalidationofagoodmanufacturingprocessforil4drivenexpansionofchimericcytokinereceptorexpressingcartcells
AT farzanehfarzin developmentandvalidationofagoodmanufacturingprocessforil4drivenexpansionofchimericcytokinereceptorexpressingcartcells
AT fisherchristopherd developmentandvalidationofagoodmanufacturingprocessforil4drivenexpansionofchimericcytokinereceptorexpressingcartcells
AT hopeandrew developmentandvalidationofagoodmanufacturingprocessforil4drivenexpansionofchimericcytokinereceptorexpressingcartcells
AT adamiantonella developmentandvalidationofagoodmanufacturingprocessforil4drivenexpansionofchimericcytokinereceptorexpressingcartcells
AT maherjohn developmentandvalidationofagoodmanufacturingprocessforil4drivenexpansionofchimericcytokinereceptorexpressingcartcells