TCR-engineered T cells and CAR T cell manufacturing on clinical scale

Engineered T cells for cancer therapy and other therapeutical applications

Chimeric antigen receptor (CAR) T cell therapy is undoubtedly leading a revolution in cancer therapy.¹ Recent successes in treating hematologic malignancies with CAR T cell therapy and the availability of several commercial products have prompted increased interest in cancer immunotherapy.² These advances have also opened new lines of research looking to optimize the safety and efficacy of CAR T cell therapies via complex gene engineering³ and promising developments like engineered T cell receptors (TCRs).⁴ More recently, genetically engineered T cells are also being applied in other indication areas like solid tumors, infectious diseases, and autoimmunity.^5-7

Overview of the CliniMACS Prodigy T Cell Transduction (TCT; above) and CliniMACS Prodigy T Cell Transduction – Large-Scale (TCT-LS; below) used in CAR T cell and TCR-engineered T cell manufacturing.

TCR-engineered T cells and CAR T cell manufacturing on the CliniMACS Prodigy®

Currently, CAR T cell and TCR T cell production consists of various complex process steps. GMP cell manufacturing is labor intensive, and as such, represents a significant challenge in further development of gene-modified T cell therapies. The CliniMACS Prodigy T Cell Transduction (TCT) and CliniMACS Prodigy T Cell Transduction – Large-Scale (TCT-LS) provide a unique all-in-one solution to this challenge. Well-established and widely used in clinical settings^8–12, the CliniMACS Prodigy Platform generates gene-modified T cells via viral transduction in a standardized and automated process.¹³ The closed system is a breakthrough in cell manufacturing designed to facilitate the transition of novel strategies like TCR T cell therapies to clinical use.⁴

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Overview of the CliniMACS Prodigy T Cell Engineering (TCE) used in complex engineered T cell manufacturing.

Non-viral T cell engineering on the CliniMACS Prodigy

The CliniMACS® Electroporator expands the cell processing capabilities of the CliniMACS Prodigy Platform to include non-viral gene-engineering in a closed and automated fashion. Electroporation parameters are flexibly adaptable to optimize transfection protocols for different gene-engineering technologies, such as TALENs, CRISPR/Cas, zinc finger nucleases, the Sleeping Beauty transposon system, siRNA, and more. The CliniMACS Prodigy T Cell Engineering (TCE) enables complex T cell engineering via electroporation and/or viral transduction, leading the way in GMP-compliant gene engineering.^14–16

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An overview of gene-engineered T cell manufacturing options with the CliniMACS Prodigy Platform

	CliniMACS Prodigy T Cell Transduction (TCT)	CliniMACS Prodigy T Cell Transduction Large-Scale (TCT-LS)	CliniMACS Prodigy T Cell Engineering (TCE)
Main application	CAR T cell manufacturing	TCR T cell manufacturing	Complex engineered T Cell manufacturing
General features	T cell isolation, activation, viral transduction, expansion, rebuffering, and harvest	T cell isolation, activation, viral transduction, expansion, rebuffering, and harvest	T cell isolation, activation, viral transduction and/or electroporation, expansion, rebuffering, and harvest
Starting material	Heterogenous blood products like whole blood, buffy coat, leukapheresis	Fresh or frozen leukapheresis	Heterogenous blood products like whole blood, buffy coat, leukapheresis
Gene engineering method	Viral transduction (lentivirus or retrovirus)	Viral transduction (lentivirus or retrovirus)	Electroporation and/or viral transduction (lentivirus or retrovirus)
Culture capacity (volume)	250 mL	600 mL	250 mL
Number of harvested T cells	~5x10⁹	~2x10¹⁰	Electroporation included: ~2x10⁹ No electroporation: ~5x10⁹

References

Zhao, L. and Cao, Y.J. (2019) Engineered T cell therapy for cancer in the clinic. Front. Immunol. 10: 2250.
Upadhaya, S. et al. (2020) Immuno-oncology drug development forges on despite COVID-19. Nature Reviews Drug Discovery 19: 751–752.
Rafiq, S. et al. (2020) Engineering strategies to overcome the current roadblocks in CAR T cell therapy. Nat Rev Clin Oncol 17: 147–167.
Zhao, Q. et al. (2021) Engineered TCR-T cell immunotherapy in anticancer precision medicine: pros and cons. Front. Immunol. 12: 658753.
Castellarin, M. et al. (2018) Driving cars to the clinic for solid tumors. Gene Ther. 25: 165–175.
Seif, M. et al. (2019) CAR T cells beyond cancer: Hope for immunomodulatory therapy of infectious diseases. Front. Immunol. 10: 2711.
Mougiakakos, D. et al. (2021) CD19-targeted CAR T cells in refractory systemic lupus erythematosus. N. Engl. J. Med. 385: 567–569.
Castella, M. et al. (2020) Point-of-care CAR T-cell production (ARI-0001) using a closed semi-automatic bioreactor: Experience from an academic phase I clinical trial. Front. Immunol. 11: 482.
Jackson, Z. et al. (2020) Automated manufacture of autologous CD19 CAR-T cells for treatment of non-Hodgkin lymphoma. Front. Immunol. 7 (11): 1941.
Maschan, M. et al. (2021) Multiple site place-of-care manufactured anti-CD19 CAR-T cells induce high remission rates in B-cell malignancy patients. Nat Commun. 12 (1): 7200.
Shah, N.N. et al. (2020) Bispecific anti-CD20, anti-CD19 CAR T cells for relapsed B cell malignancies: a phase 1 dose escalation and expansion trial. Nat. Med. 26: 1569–1575.
Ortiz-Maldonado, V. et al. (2020). CART19-BE-01: A multicenter trial of ARI-001 cell therapy in patients with CD19+ relapsed/refractory malignancies. Mol Ther. 29 (2): 636–664.
Lock, D. et al. (2017) Automated manufacturing of potent CD20-directed chimeric antigen receptor T cells for clinical use. Hum. Gene Ther. 10: 914–925.
Schwarze, L. I. et al. (2021) Automated production of CCR5-negative CD4+ T cells in a GMP-compatible, clinical-scale for treatment of HIV-positive patients. Gene Ther. 28 (9 ):572–587
Alzubi, J. et al. (2020) Automated generation of gene-edited CAR T cells at clinical scale. Mol. Ther. Methods Clin. Dev. 20: 379–388.
Lock, D. et al. (2022) Automated, scaled, transposon-based production of CAR T cells. J. Immunotherapy Cancer 10 (9): e005189.

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