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Definition and application of good manufacturing process-compliant production of CEA-specific chimeric antigen receptor expressing T-cells for phase I/II clinical trial

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Abstract

Adoptive cell therapy employing gene-modified T-cells expressing chimeric antigen receptors (CARs) has shown promising preclinical activity in a range of model systems and is now being tested in the clinical setting. The manufacture of CAR T-cells requires compliance with national and European regulations for the production of medicinal products. We established such a compliant process to produce T-cells armed with a first-generation CAR specific for carcinoembryonic antigen (CEA). CAR T-cells were successfully generated for 14 patients with advanced CEA+ malignancy. Of note, in the majority of patients, the defined procedure generated predominantly CD4+ CAR T-cells with the general T-cell population bearing an effector–memory phenotype and high in vitro effector function. Thus, improving the process to generate less-differentiated T-cells would be more desirable in the future for effective adoptive gene-modified T-cell therapy. However, these results confirm that CAR T-cells can be generated in a manner compliant with regulations governing medicinal products in the European Union.

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Abbreviations

7-AAD:

7-Aminoactinomycin D

ATMP:

Advanced therapy medicinal product

CAR:

Chimeric antigen receptor

CEA:

Carcinoembryonic antigen

CTA:

Clinical trials authorization

DC:

Dendritic cell

DMSO:

Dimethyl sulfoxide

FBS:

Fetal bovine serum

GMP:

Good manufacturing process

HEPES:

4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid

HSA:

Human serum albumin

PBS:

Phosphate-buffered saline

PQ:

Process quantification

scFv:

Single-chain variable fragment

TSE:

Transmissible spongiform encephalitis

UK:

United Kingdom

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Acknowledgments

The authors wish to acknowledge the support of Cancer Research UK for this work. This work was also supported by funding from the Kay Kendall Leukaemia Fund, the FP6 Program ATTACK, the NHSBT, Christie Hospital NHS Trust, and the BBSRC (HG).

Conflict of interest

Ryan D Guest, Robert E Hawkins, and David E Gilham are cofounders of Cellular Therapeutics Ltd. All other authors do not have any conflict of interest to declare.

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Authors and Affiliations

  1. Cellular Therapeutics, Grafton Street, Manchester, M13 9XX, UK

    Ryan D. Guest, Natalia Kirillova & Sam Mowbray

  2. Clinical and Experimental Immunotherapy Group, Department of Medical Oncology, Manchester Academic Health Science Centre, Institute of Cancer Sciences, University of Manchester, Wilmslow Road, Withington, Manchester, M20 4BX, UK

    Hannah Gornall, Fiona Thistlethwaite, Robert E. Hawkins & David E. Gilham

  3. Clinical Immune and Molecular Monitoring Laboratory, Clinical and Experimental Pharmacology Group, CRUK Manchester Institute, Manchester, UK

    Dominic G. Rothwell

  4. Targeted Therapy Group, Institute of Cancer Sciences, Manchester, UK

    Eleanor J. Cheadle

  5. Stem Cells and Immunotherapy, NHSBT Liverpool Centre, Speke, Liverpool, UK

    Eric Austin & Keith Smith

  6. Stem Cell Research, NHS Blood and Transplant Oxford Centre, University of Oxford, Oxford, UK

    Suzanne M. Watt

  7. EUFETS GmbH, Vollmersbachstr. 66, 55743, Idar-Oberstein, Germany

    Klaus Kühlcke

  8. Cancer Research UK Drug Development Office, Angel Building, 407 St John Street, London, EC1V 4AD, UK

    Nigel Westwood

  9. Department of Medical Oncology, Christie Hospital NHS Trust, Wilmslow Road, Withington, Manchester, UK

    Fiona Thistlethwaite & Robert E. Hawkins

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  1. Ryan D. Guest
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  2. Natalia Kirillova
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  13. Robert E. Hawkins
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  14. David E. Gilham
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Corresponding author

Correspondence to David E. Gilham.

Additional information

Robert E. Hawkins and David E. Gilham are Joint Senior Authors.

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Guest, R.D., Kirillova, N., Mowbray, S. et al. Definition and application of good manufacturing process-compliant production of CEA-specific chimeric antigen receptor expressing T-cells for phase I/II clinical trial. Cancer Immunol Immunother 63, 133–145 (2014). https://doi.org/10.1007/s00262-013-1492-9

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