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Manufacturing Chimeric Antigen Receptor CAR T Cells for Adoptive Immunotherapy
Published on: December 17, 2019
Next-Generation Manufacturing Protocols Enriching TSCM CAR T Cells Can Overcome Disease-Specific T Cell Defects in
Silvia Arcangeli1, Laura Falcone1, Barbara Camisa1,2
1Innovative Immunotherapies Unit, Division of Immunology, Transplantation and Infectious Diseases, IRCCS San Raffaele Scientific Institute, Milan, Italy.
Abstract:
Chimeric antigen receptor (CAR) T cell expansion and persistence emerged as key efficacy determinants in cancer patients. These features are typical of early-memory T cells, which can be enriched with specific manufacturing procedures, providing signal one and signal two in the proper steric conformation and in the presence of homeostatic cytokines. In this project, we exploited our expertise with paramagnetic beads and IL-7/IL-15 to develop an optimized protocol for CAR T cell production based on reagents, including a polymeric nanomatrix, which are compatible with automated manufacturing via the CliniMACS Prodigy. We found that both procedures generate similar CAR T cell products, highly enriched of stem cell memory T cells (TSCM) and equally effective in counteracting tumor growth in xenograft mouse models. Most importantly, the optimized protocol was able to expand CAR TSCM from B-cell acute lymphoblastic leukemia (B-ALL) patients, which in origin were highly enriched of late-memory and exhausted T cells. Notably, CAR T cells derived from B-ALL patients proved to be as efficient as healthy donor-derived CAR T cells in mediating profound and prolonged anti-tumor responses in xenograft mouse models. On the contrary, the protocol failed to expand fully functional CAR TSCM from patients with pancreatic ductal adenocarcinoma, suggesting that patient-specific factors may profoundly affect intrinsic T cell quality. Finally, by retrospective analysis of in vivo data, we observed that the proportion of TSCM in the final CAR T cell product positively correlated with in vivo expansion, which in turn proved to be crucial for achieving long-term remissions. Collectively, our data indicate that next-generation manufacturing protocols can overcome initial T cell defects, resulting in TSCM-enriched CAR T cell products qualitatively equivalent to the ones generated from healthy donors. However, this positive effect may be decreased in specific conditions, for which the development of further improved protocols and novel strategies might be highly beneficial.
Insights
Next-generation manufacturing optimizes chimeric antigen receptor (CAR) T cell production, enriching stem cell memory T cells (Tscm) for potent anti-cancer activity. This approach overcomes patient-specific T cell defects, improving CAR T cell therapy efficacy.
Area of Science:
- Immunology
- Cell Therapy
- Biotechnology
Background:
- Chimeric antigen receptor (CAR) T cell expansion and persistence are critical for cancer treatment efficacy.
- Early-memory T cells, particularly stem cell memory T cells (Tscm), are key for sustained CAR T cell function.
- Current manufacturing methods aim to enrich these memory T cell populations.
Purpose of the Study:
- To develop an optimized protocol for CAR T cell production using paramagnetic beads, IL-7/IL-15, and a polymeric nanomatrix.
- To assess the compatibility of the optimized protocol with automated manufacturing systems (CliniMACS Prodigy).
- To evaluate the efficacy of CAR Tscm generated from both healthy donors and cancer patients.
Main Methods:
- Utilized expertise in paramagnetic beads and cytokines (IL-7/IL-15) to create an optimized CAR T cell manufacturing protocol.
- Incorporated a polymeric nanomatrix compatible with automated CliniMACS Prodigy manufacturing.
- Compared the optimized protocol with existing procedures, assessing Tscm enrichment and in vitro/in vivo anti-tumor activity.
Main Results:
- Both optimized and standard procedures yielded CAR T cell products rich in Tscm, showing equal efficacy in preclinical models.
- The optimized protocol successfully expanded functional CAR Tscm from B-cell acute lymphoblastic leukemia (B-ALL) patients, even from T cells initially enriched in late-memory/exhausted cells.
- CAR Tscm from B-ALL patients demonstrated efficacy comparable to those from healthy donors; however, the protocol failed for pancreatic cancer patients, indicating patient-specific limitations.
Conclusions:
- Next-generation manufacturing protocols can enhance CAR T cell quality by enriching Tscm, potentially overcoming initial T cell defects.
- The proportion of Tscm in CAR T cell products correlates positively with in vivo expansion and long-term remission.
- While effective for some patient groups, further protocol improvements are needed for conditions where patient-specific factors limit CAR T cell function.
