Production of CAR T-cells by GMP-grade lentiviral vectors: latest advances and future prospects

Poorebrahim, Mansour; Sadeghi, Sedigheh; Fakhr, Elham; Abazari, Mohammad Foad; Poortahmasebi, Vahdat; Kheirollahi, Asma; Askari, Hasan; Rajabzadeh, Aliakbar; Rastegarpanah, Malihe; Linē, Aija; Cid-Arregui, Ángel

doi:10.1080/10408363.2019.1633512

Cited by 62 publications

(45 citation statements)

References 174 publications

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“…A ficoll density gradient centrifugation is used for the removal of granulocytes, red blood cells, and platelets [45]. Alternatively, automated cell-washers can be employed to isolate T cells [46]. Further instruments have been developed to facilitate or combine the manufacturing steps in one system, for example, the Sefia™ Cell Processing System for isolation, harvesting, and final formulation of cellular products or the CliniMACS Prodigy ® for automated GMP-compliant manufacturing of various cell types [47,48].…”

Section: Isolation and Enrichment Of T Vellsmentioning

confidence: 99%

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Optimizing Manufacturing Protocols of Chimeric Antigen Receptor T Cells for Improved Anticancer Immunotherapy

Stock

Schmitt

Sellner

2019

IJMS

112

106

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Chimeric antigen receptor (CAR) T cell therapy can achieve outstanding response rates in heavily pretreated patients with hematological malignancies. However, relapses occur and they limit the efficacy of this promising treatment approach. The cellular composition and immunophenotype of the administered CART cells play a crucial role for therapeutic success. Less differentiated CART cells are associated with improved expansion, long-term in vivo persistence, and prolonged anti-tumor control. Furthermore, the ratio between CD4+ and CD8+ T cells has an effect on the anti-tumor activity of CART cells. The composition of the final cell product is not only influenced by the CART cell construct, but also by the culturing conditions during ex vivo T cell expansion. This includes different T cell activation strategies, cytokine supplementation, and specific pathway inhibition for the differentiation blockade. The optimal production process is not yet defined. In this review, we will discuss the use of different CART cell production strategies and the molecular background for the generation of improved CART cells in detail.

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Section: Isolation and Enrichment Of T Vellsmentioning

confidence: 99%

“…The two most commonly used strategies for CART cell production are based on either IL-2 or IL-7, with or without IL-15. So far, IL-2 was primarily used for T cell expansion in previous clinical studies [3,45,46]. For example, IL-2 is supplemented for the production of Yescarta ® (Axicabtagene Ciloleucel) [70].…”

Section: Stimulation With Cytokinesmentioning

confidence: 99%

Optimizing Manufacturing Protocols of Chimeric Antigen Receptor T Cells for Improved Anticancer Immunotherapy

Stock

Schmitt

Sellner

2019

IJMS

112

106

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“…Retroviral and lentiviral vectors are costly and cumbersome to produce in large batches, and their use requires that specific quality control assays regarding the presence of replicationcompetent retrovirus (RCR) are performed in the final product. 13 Moreover, use of retroviral vectors requires preactivation of T cells, which generally adds at least 2 days to the manufacturing process. In combination with the current methods of T cell expansion, like Wave bioreactors, or G-REX flask, total production time ranges from 12 to 16 days.…”

Section: Introductionmentioning

confidence: 99%

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

et al. 2020

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Recently approved by the FDA and European Medicines Agency, CART cell therapy is a new treatment option for B-cell malignancies. Currently, CART cells are manufactured in centralized facilities and face bottlenecks like complex scaling up, high costs, and logistic operations. These difficulties are mainly related to the use of viral vectors and the requirement to expand CART cells to reach the therapeutic dose. In this paper, by using Sleeping Beauty-mediated genetic modification delivered by electroporation, we show that CART cells can be generated and used without the need for ex vivo activation and expansion, consistent with a point-of-care (POC) approach. Our results show that minimally manipulated CART cells are effective in vivo against RS4;11 leukemia cells engrafted in NSG mice even when inoculated after only 4 h of gene transfer. In an effort to better characterize the infused CART cells, we show that 19BBz T lymphocytes infused after 24 h of electroporation (where CAR expression is already detectable) can improve the overall survival and reduce tumor burden in organs of mice engrafted with RS4;11 or Nalm-6 B cell leukemia. A side-by-side comparison of POC approach with a conventional 8-day expansion protocol using Transact beads demonstrated that both approaches have equivalent antitumor activity in vivo. Our data suggest that POC approach is a viable alternative for the generation and use of CART cells, overcoming the limitations of current manufacturing protocols. Its use has the potential to expand CAR immunotherapy to a higher number of patients, especially in the context of low-income countries.

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“…Regardless of impressive efficacy of reinvigorated CAR T cells, a risk assessment of unexpected toxicities has remained to be investigated [11]. "On-target, on-tumor" toxicity can be a direct consequence of reinvigorated CAR T cells, which is related to the induction of CRS and tumor lysis syndrome induced by abundant cytokine release and excessive tumor cell death in tumor site [131]. On the other hand, a catastrophic and rapid "on-target, off-tumor" toxicity might be observed after infusion of armored CAR T cells targeting a tumor-associated antigen (TAA), which is expressed in both tumor and normal cells.…”

Section: Safety Considerations and Concluding Remarksmentioning

confidence: 99%

Counteracting CAR T cell dysfunction

et al. 2021

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In spite of high rates of complete remission following chimeric antigen receptor (CAR) T cell therapy, the efficacy of this approach is limited by generation of dysfunctional CAR T cells in vivo, conceivably induced by immunosuppressive tumor microenvironment (TME) and excessive antigen exposure. Exhaustion and senescence are two critical dysfunctional states that impose a pivotal hurdle for successful CAR T cell therapies. Recently, modified CAR T cells with an “exhaustion-resistant” phenotype have shown superior antitumor functions and prolonged lifespan. In addition, several studies have indicated the feasibility of senescence delay in CAR T cells. Here, we review the latest reports regarding blockade of CAR T cell exhaustion and senescence with a particular focus on the exhaustion-inducing pathways. Subsequently, we describe what potential these latest insights offer for boosting the potency of adoptive cell transfer (ACT) therapies involving CAR T cells. Furthermore, we discuss how induction of costimulation, cytokine exposure, and TME modulation can impact on CAR T cell efficacy and persistence, while potential safety issues associated with reinvigorated CAR T cells will also be addressed.

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Production of CAR T-cells by GMP-grade lentiviral vectors: latest advances and future prospects

Cited by 62 publications

References 174 publications

Optimizing Manufacturing Protocols of Chimeric Antigen Receptor T Cells for Improved Anticancer Immunotherapy

Optimizing Manufacturing Protocols of Chimeric Antigen Receptor T Cells for Improved Anticancer Immunotherapy

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

Counteracting CAR T cell dysfunction

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