Development of a cGMP-compliant process to manufacture donor-derived, CD45RA-depleted memory CD19-CAR T cells

Autologous chimeric antigen receptor (CAR) T cells targeting the CD19 antigen have demonstrated a high complete response rate in relapsed/refractory B-cell malignancies. However, autologous CAR T cell therapy is not an option for all patients. Here we optimized conditions for clinical-grade manufact...

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Bibliographic Details
Published in:Gene therapy 2023-04, Vol.30 (3-4), p.222-231
Main Authors: Kim-Hoehamer, Young-In, Riberdy, Janice M, Zheng, Fei, Park, Jeoungeun J, Shang, Na, Métais, Jean-Yves, Lockey, Timothy, Willis, Catherine, Akel, Salem, Moore, Jennifer, Meagher, Michael M, Velasquez, M Paulina, Triplett, Brandon M, Talleur, Aimee C, Gottschalk, Stephen, Zhou, Sheng
Format: Article
Language:English
Subjects:
Antigens
Antigens, CD19 - genetics
Antitumor activity
CD19 antigen
CD4 antigen
CD45RA antigen
Cell culture
Cell therapy
Chimeric antigen receptors
Cryopreservation
Cyclic GMP - metabolism
Good Manufacturing Practice
Humans
Immunological memory
Immunotherapy, Adoptive - methods
Lymphocytes
Lymphocytes B
Lymphocytes T
Malignancy
Manufacturing
Memory cells
Neoplasms
Phenotypes
Receptors, Antigen, T-Cell
T-Lymphocytes
Tissue engineering
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Summary:Autologous chimeric antigen receptor (CAR) T cells targeting the CD19 antigen have demonstrated a high complete response rate in relapsed/refractory B-cell malignancies. However, autologous CAR T cell therapy is not an option for all patients. Here we optimized conditions for clinical-grade manufacturing of allogeneic CD19-CAR T cells using CD45RA-depleted donor memory T cells (Tm) for a planned clinical trial. Tm were activated using the MACS GMP T Cell TransAct reagent and transduced in the presence of LentiBOOST with a clinical-grade lentiviral vector that encodes a 2nd generation CD19-CAR with a 41BB.zeta endodomain. Transduced T cells were transferred to a G-Rex cell culture device for expansion and harvested on day 7 or 8 for cryopreservation. The resulting CD19-CAR(Mem) T cells expanded on average 34.2-fold, and mean CAR expression was 45.5%. The majority of T cells were CD4 and had a central memory or effector memory phenotype, and retained viral specificity. CD19-CAR(Mem) T cells recognized and killed CD19-positive target cells in vitro and had potent antitumor activity in an ALL xenograft model. Thus we have successfully developed a current good manufacturing practice-compliant process to manufacture donor-derived CD19-CAR(Mem) T cells. Our manufacturing process could be readily adapted for CAR(Mem) T cells targeting other antigens.
ISSN:0969-7128
1476-5462
DOI:10.1038/s41434-021-00307-0