Streamlined production of genetically modified T cells with activation, transduction and expansion in closed-system G-Rex bioreactors

Gagliardi, Christine; Khalil, Mariam; Foster, Aaron E.

doi:10.1016/j.jcyt.2019.10.006

Cited by 15 publications

(14 citation statements)

References 18 publications

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“…This bioreactor can be placed in a regular lab incubator; for media changes and cell harvests, the bioreactor is opened in a biosafety cabinet, to manually pipette fluids and cells in and out under aseptic conditions. Recently, a functionally closed version of this system was developed, allowing for a peristaltic pump to move fluids and cells in and out of the bioreactor (73,74). A single unit of this bioreactor can provide a 500 cm 2 gas-permeable surface area with 5 L media capacity, which is sufficient to expand large numbers of CAR T cells for clinical applications.…”

Section: The Evolution Of Car T Cell Manufacturing Platformsmentioning

confidence: 99%

Scalable Manufacturing of CAR T Cells for Cancer Immunotherapy

Abou‐El‐Enein

Elsallab

Feldman³

et al. 2021

Blood Cancer Discovery

122

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As of April 2021, there are five commercially available chimeric antigen receptor (CAR) T cell therapies for hematologic malignancies. With the current transition of CAR T cell manufacturing from academia to industry, there is a shift toward Good Manufacturing Practice (GMP)-compliant closed and automated systems to ensure reproducibility and to meet the increased demand for patients with cancer. In this review, we describe current CAR T cell clinical manufacturing models and discuss emerging technologic advances that embrace scaling and production optimization. We summarize measures being used to shorten CAR T cell manufacturing times and highlight regulatory challenges to scaling production for clinical use. Significance:As the demand for CAR T cell cancer therapy increases, several closed and automated production platforms are being deployed, and others are in development. This review provides a critical appraisal of these technologies, which can be leveraged to scale and optimize the production of nextgeneration CAR T cells. iNtRODUctiONChimeric antigen receptor (CAR)-modified T cells have emerged as an efficacious treatment for patients with certain hematologic malignancies (1). Currently, five CAR T cell products are approved for commercial use and available on the U.S. market: three for B-cell leukemia and lymphoma (tisagenlecleucel, axicabtagene ciloleucel, lisocabtagene maraleucel), one for mantle cell lymphoma (brexucabtagene autoleucel), and one for multiple myeloma (idecabtagene vicleucel; refs. 2-8). These therapies involve genetically modifying patient-derived (autologous) peripheral blood T cells to express a CAR directed against antigens present on the surface of targeted tumor cells, such as the CD19 molecule, or, in the case of idecabtagene vicleucel, B-cell maturation antigen (BCMA;. After antigen recognition, the intracellular signaling domains activate the immune effector and memory functions of the CAR T cells. Once activated, these T cells proliferate, infiltrate tumor sites, secrete cytokines, and release cytolytic granules to eliminate targeted cells in an antigen-dependent manner (1). All approved CAR T cell products are second-generation CARs that incorporate CD28 or CD137 (4-1BB) costimulatory signals, which are essential for eliciting a clinically relevant immune response (9-15). These CAR T cells were able to elicit complete responses (CR) in 32% to 67% of patients with lymphoma and showed better CR rates in patients with leukemia (16)(17)(18)(19).The increasing success of CAR T immunotherapies in relapsed/refractory hematologic malignancies sparked the interest of pharmaceutical companies, and several products targeting a variety of cancers are currently in the pipeline (20). However, many limitations to current CAR T cell manufacturing must be overcome before this modality can be fully integrated into routine clinical practice. First, most CAR T cell trials to date have used autologous peripheral blood and apheresis as the main cell sources for manufacturing

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Section: The Evolution Of Car T Cell Manufacturing Platformsmentioning

confidence: 99%

Scalable Manufacturing of CAR T Cells for Cancer Immunotherapy

Abou‐El‐Enein

Elsallab

Feldman³

et al. 2021

Blood Cancer Discovery

122

View full text Add to dashboard Cite

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“…This, together with the efficient gas exchange and ready availability of abundant nutrients in culture medium, may explain the much more rapid and efficient expansion in G-Rex compared with T-flasks. G-Rex has been used by other groups for the optimal expansion of several types of blood cells, including tumor-infiltrating lymphocytes (TILs) [19,20], antigen-specific T cells [8,12,17,[33][34][35], gd T cells [23], Tregs [21,22], NKs [15], CIKs [25], CAR-Ts [16,24], megakaryocytes [36] and red blood cells [37] (reviewed in [9]). In most of these published cases, medium addition or exchange, as well as culture splitting, was performed with cell counting at various phases of the culture to define optimal dilution and collection times.…”

Section: Discussionmentioning

confidence: 99%

“…G-Rex devices are singleuse, good manufacturing practices (GMP)-compliant medical devices that are easily accommodated in standard 5% CO 2 incubators and come in scalable formats with 10-, 100-or 500-cm 2 membranes and a constant ratio of maximum volume-to-membrane surface area [8,13,14]. The G-Rex devices have been optimized for expansion of different types of T cells for research or clinical use [10,13,15,16]. Thus G-Rex devices have been used to produce antigen-specific T cells [8,12,17,18], tumor-infiltrating lymphocytes (TILs) [19,20], regulatory T cells (Tregs) [21,22], g/d T cells [23] and chimeric antigen receptor (CAR) T cells [16,24] as well as CIKs [25].…”

Section: Introductionmentioning

confidence: 99%

“…The G-Rex devices have been optimized for expansion of different types of T cells for research or clinical use [10,13,15,16]. Thus G-Rex devices have been used to produce antigen-specific T cells [8,12,17,18], tumor-infiltrating lymphocytes (TILs) [19,20], regulatory T cells (Tregs) [21,22], g/d T cells [23] and chimeric antigen receptor (CAR) T cells [16,24] as well as CIKs [25].…”

Section: Introductionmentioning

confidence: 99%

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Optimization of therapeutic T cell expansion in G-Rex device and applicability to large-scale production for clinical use

et al. 2022

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“…Additionally, since centralized production of T-cell products is expensive, it is plausible that smaller production facilities could be generated on a franchise-like basis where vectors and cell culture materials are supplied to the local T-cell production sites. In this direction, “bioreactors” (i.e., smaller contained production units) are being investigated in clinical trials [ 105 , 106 ]. Theoretically, this expansion of TCR therapies may increase the success rate, yet it will require the training of highly specialized personnel, the establishment of Good Manufacturing Practices-certified facilities, and conformation to the same stringent FDA regulations that surround the production of TCR products.…”

Section: Introductionmentioning

confidence: 99%

T-cell receptor-based therapy: an innovative therapeutic approach for solid tumors

Tsimberidou

Morris

et al. 2021

J Hematol Oncol

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T-cell receptor (TCR)-based adoptive therapy employs genetically modified lymphocytes that are directed against specific tumor markers. This therapeutic modality requires a structured and integrated process that involves patient screening (e.g., for HLA-A*02:01 and specific tumor targets), leukapheresis, generation of transduced TCR product, lymphodepletion, and infusion of the TCR-based adoptive therapy. In this review, we summarize the current technology and early clinical development of TCR-based therapy in patients with solid tumors. The challenges of TCR-based therapy include those associated with TCR product manufacturing, patient selection, and preparation with lymphodepletion. Overcoming these challenges, and those posed by the immunosuppressive microenvironment, as well as developing next-generation strategies is essential to improving the efficacy and safety of TCR-based therapies. Optimization of technology to generate TCR product, treatment administration, and patient monitoring for adverse events is needed. The implementation of novel TCR strategies will require expansion of the TCR approach to patients with HLA haplotypes beyond HLA-A*02:01 and the discovery of novel tumor markers that are expressed in more patients and tumor types. Ongoing clinical trials will determine the ultimate role of TCR-based therapy in patients with solid tumors.

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Streamlined production of genetically modified T cells with activation, transduction and expansion in closed-system G-Rex bioreactors

Cited by 15 publications

References 18 publications

Scalable Manufacturing of CAR T Cells for Cancer Immunotherapy

Scalable Manufacturing of CAR T Cells for Cancer Immunotherapy

Optimization of therapeutic T cell expansion in G-Rex device and applicability to large-scale production for clinical use

T-cell receptor-based therapy: an innovative therapeutic approach for solid tumors

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