Characterization of clinical grade CD19 chimeric antigen receptor T cells produced using automated CliniMACS prodigy system

Abstract

BackgroundChimeric antigen receptor (CAR) T-cell therapy is highly effective for treating acute lymphoblastic leukemia and non-Hodgkin’s lymphoma with high rate complete responses. However, the broad clinical application of CAR T-cell therapy has been challenging, largely due to the lack of widespread ability to produce and high cost of CAR T-cell products using traditional methods of production. Automated cell processing in a closed system has emerged as a potential method to increase the feasibility of producing CAR T cells locally at academic centers due to its minimal reliance on experienced labor, thereby making the process less expensive and more consistent than traditional methods of production.MethodIn this study, we describe the successful production of clinical grade CD19 CAR T cells using the Miltenyi CliniMACS Prodigy Automated Cell Processor at University of Colorado Anschutz Medical Campus in a rapid manner with a high frequent CD19 CAR expression.ResultsThe final CAR T-cell product is highly active, low in immune suppression, and absent in exhaustion. Full panel cytokine assays also showed elevated production of Th1 cytokines upon IL-2 stimulation when specifically killing CD19+ target cells.ConclusionThese results demonstrate the feasibility of producing CAR T cells locally in a university hospital setting using automated cell processor for future clinical applications.

Demonstrating the Manufacture of Human CAR‐T Cells in an Automated Stirred‐Tank Bioreactor

“…[ 28 ] Results reported in ref. [27] showed a lower fold expansion in the CliniMACS Prodigy compared to those presented in this study. However, the lack of nutrients during culture seen here suggest that once the feeding strategy in the stirred bioreactor has been optimized or even switched from fed‐batch to perfusion mode, it would lead to an improved performance.…”

“…Moreover, the fold expansion at 300 rpm was 25.70 ± 1.21, which was higher than the one reported in the CliniMACS Prodigy. [ 27 ] If it is assumed an ≈40% CAR‐T positive population at the end of the run, this result would be 5 × 10 8 CAR positive viable T‐cells at harvest (for a transduction efficiency as low as 20.5%, there would still be 2.56 × 10 8 CAR positive viable T‐cells at harvest). These numbers fall in the range of target doses currently administered to patients in the FDA approved CAR‐T therapies ( Table 1 ), making a small‐scale, automated STR potentially suitable for the manufacturing of personalized medicines.…”

Mechanisms of resistance to CAR T cell therapy

“…Iterative modifications to the manufacturing process have the potential to improve the efficacy of CAR T cells, which will probably be of greater importance as CAR T cells are tested beyond B cell malignancies. Novel approaches to CAR T cell manufacturing, such as use of closed-system automated devices, incorporating the aforementioned foundations are just beginning to be implemented 32,33 ; clinical trials using these approaches and further analysis of how these techniques will impact future CAR T cell therapy are warranted. Even if a CAR T cell product can be manufactured successfully, the starting T cell phenotype has been demonstrated to be an important determinant of subsequent clinical activity.…”

Development of CAR-T cell therapy for B-ALL using a point-of-care approach

“…The use of automated CAR-T cell production equipment (for example, Miltenyi Prodigy) will likely improve the process, but the high upfront costs of acquisition and lack of customization might hamper their use in the short term. 37,38 Meanwhile, preclinical and clinical data are showing that T cells with a less differentiated phenotype, such as naïve or central memory T cells, have improved antitumor responses in vivo. [39][40][41] In fact, a recent report described that the presence of a memory-like CD27+CD45RO-CD8+T cell subpopulation was associated with improved responses in patients with chronic lymphocytic leukemia treated with anti-CD19 CAR-T cells.…”