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Updated: Jun 15, 2025
Manufacturing Chimeric Antigen Receptor CAR T Cells for Adoptive Immunotherapy
Published on: December 17, 2019
Next-generation BCMA-targeted chimeric antigen receptor CARTemis-1: the impact of manufacturing procedure on CAR
Belén Sierro-Martínez1, Virginia Escamilla-Gómez1, Laura Pérez-Ortega1
1Instituto de Biomedicina de Sevilla, IBiS/Hospital Universitario Virgen del Rocío/CSIC/Universidad de Sevilla, Sevilla, Spain.
Purpose:
CAR therapy targeting BCMA is under investigation as treatment for multiple myeloma. However, given the lack of plateau in most studies, pursuing more effective alternatives is imperative. We present the preclinical and clinical validation of a new optimized anti-BCMA CAR (CARTemis-1). In addition, we explored how the manufacturing process could impact CAR-T cell product quality and fitness.
Methods:
CARTemis-1 optimizations were evaluated at the preclinical level both, in vitro and in vivo. CARTemis-1 generation was validated under GMP conditions, studying the dynamics of the immunophenotype from leukapheresis to final product. Here, we studied the impact of the manufacturing process on CAR-T cells to define optimal cell culture protocol and expansion time to increase product fitness.
Results:
Two different versions of CARTemis-1 with different spacers were compared. The longer version showed increased cytotoxicity. The incorporation of the safety-gene EGFRt into the CARTemis-1 structure can be used as a monitoring marker. CARTemis-1 showed no inhibition by soluble BCMA and presents potent antitumor effects both in vitro and in vivo. Expansion with IL-2 or IL-7/IL-15 was compared, revealing greater proliferation, less differentiation, and less exhaustion with IL-7/IL-15. Three consecutive batches of CARTemis-1 were produced under GMP guidelines meeting all the required specifications. CARTemis-1 cells manufactured under GMP conditions showed increased memory subpopulations, reduced exhaustion markers and selective antitumor efficacy against MM cell lines and primary myeloma cells. The optimal release time points for obtaining the best fit product were > 6 and < 10 days (days 8-10).
Conclusions:
CARTemis-1 has been rationally designed to increase antitumor efficacy, overcome sBCMA inhibition, and incorporate the expression of a safety-gene. The generation of CARTemis-1 was successfully validated under GMP standards. A phase I/II clinical trial for patients with multiple myeloma will be conducted (EuCT number 2022-503063-15-00).
Insights
This study introduces CARTemis-1, an optimized chimeric antigen receptor (CAR) T-cell therapy for multiple myeloma. CARTemis-1 demonstrates potent anti-tumor effects and successful GMP validation, paving the way for clinical trials.
Area of Science:
- Immunotherapy
- Oncology
- Cellular Therapy
Background:
- Chimeric antigen receptor (CAR) T-cell therapy targeting BCMA is a promising treatment for multiple myeloma.
- Limitations in current CAR T-cell therapies necessitate the development of more effective alternatives.
- Optimizing CAR T-cell manufacturing processes is crucial for enhancing product quality and therapeutic efficacy.
Purpose of the Study:
- To present the preclinical and clinical validation of an optimized anti-BCMA CAR T-cell therapy, named CARTemis-1.
- To investigate the impact of manufacturing processes on CAR-T cell product quality and fitness.
- To define optimal cell culture protocols and expansion times for enhanced CAR-T cell product fitness.
Main Methods:
- CARTemis-1 optimizations were evaluated in vitro and in vivo preclinical models.
- CARTemis-1 generation was validated under Good Manufacturing Practice (GMP) conditions, monitoring immunophenotype dynamics.
- Comparative analysis of IL-2 versus IL-7/IL-15 expansion methods was performed to assess CAR-T cell proliferation, differentiation, and exhaustion.
Main Results:
- A longer spacer version of CARTemis-1 demonstrated increased cytotoxicity and potent anti-tumor effects, overcoming soluble BCMA inhibition.
- Expansion with IL-7/IL-15 resulted in greater proliferation, reduced differentiation, and less exhaustion compared to IL-2.
- GMP-manufactured CARTemis-1 exhibited increased memory subpopulations, reduced exhaustion markers, and selective efficacy against multiple myeloma cells, with optimal release at days 8-10.
Conclusions:
- CARTemis-1 is rationally designed for enhanced anti-tumor efficacy, resistance to sBCMA inhibition, and includes a safety gene (EGFRt).
- The generation of CARTemis-1 was successfully validated under GMP standards.
- A Phase I/II clinical trial for multiple myeloma patients is planned to further evaluate CARTemis-1 efficacy and safety.
![Dynamic Imaging of Chimeric Antigen Receptor T Cells with [18F]Tetrafluoroborate Positron Emission Tomography/Computed Tomography](/_next/image?url=https%3A%2F%2Fcloudfront.jove.com%2FCDNSource%2Fteasers%2F62334.jpg&w=3840&q=50)