Persistent challenges in cell therapy manufacturing necessitate the implementation and utilization of innovative technologies. In this blog post, we will highlight a next-generation cell therapy manufacturing process that shortens the time to manufacture autologous CAR-T cells from a typical 7-14-day timeline to a 24-hour process. This advanced, automated, closed, lentivirus-based method allows fewer manual touchpoints and improves efficiency. By enabling improved patient access, the decentralized manufacturing model holds the key to overcoming barriers and expanding the reach of life-saving treatments.
Point of care cell therapy: An emerging concept for decentralized manufacturing
Conventional pharmacological manufacturing is a highly centralized process and involves complicated supply-chain logistics and cryopreservation. While this is a time-tested and economically sustainable process for traditional pharmacological drugs, it may be difficult to scale, especially with autologous cell therapies.
Most cell therapy innovations were pioneered by clinicians near the patient-bedsides at academic health centers (1). These therapies were initially designed for a handful of patients, however, when it comes to manufacturing, there are significant challenges with scaling these processes.
Centralized cell therapy manufacturing can be time-consuming due to several factors. Firstly, the process of isolating and expanding cells can take a significant amount of time. The median time for traditional CAR-T cell manufacturing can be over 20 days (2-4).
In addition to the manufacturing process, ensuring the therapy’s accessibility to patients can be a time-consuming challenge, especially for those far from centralized manufacturing facilities. The logistics involved in transporting the therapy to these patients can pose significant hurdles and delays.
Decentralization shifts cell therapy manufacturing to a more distributed process by bringing it closer to the patients. One of the approaches is introducing point of care facilities with the required infrastructure to support production, thus reducing the turnaround time (5).
Innovation in automation, and the use of closed and modular systems, which streamline cell therapy manufacturing processes help to make this feasible. These technologies enable patient cell processing in a localized setting, while providing lower logistical costs associated with transportation and storage of patient cells.
Thus, in the long run, decentralization of autologous cell therapies can help enable faster and wider access to treatment and therefore help improve patient outcomes.
A shortened CAR-T cell manufacturing workflow using a lentivirus-based gene editing method
Recent studies have highlighted novel T cell manufacturing platforms and processes with a shortened ex-vivo expansion step reduces overall time to manufacture, while producing CAR-T cells that have improved anti-tumor activity in preclinical and early clinical settings (6–8). The ability to retain less differentiated, early memory and naive central memory T cell populations for therapeutic delivery and to aim for a multi-functional T cell pool expansion in patients in vivo, set these new workflows apart from classical cell manufacturing.
In a recent study, scientists at Thermo Fisher Scientific leveraged the Gibco™ CTS™ Detachable Dynabeads™ CD3/CD28 magnetic beads with the Gibco™ CTS ™ DynaCellect™ Magnetic Separation System for one-step isolation and activation of T cells in a 24-hour lentiviral (LV)-based CAR-T workflow. These steps are integrated via closed, automated instrumentation and software, reducing the manual touchpoints.
Listed below are the key steps:
1. Isolation and activation: CTS™ Detachable Dynabeads CD3/CD28 magnetic beads with the CTS™ DynaCellect Magnetic Separation System were used to isolate T cells from quarter Leukopaks. The active-release CTS™ Detachable Dynabeads CD3/CD28 magnetic beads can be removed actively at any stage, which allows for greater control of the process and reduces cell death due to overactivation and exhaustion of T cells as seen with passive release. The one-step isolation process yields highly pure T cells population.
2. Lentiviral transduction: Post-isolation cells were infected with a lentiviral vector with a CD-19 CAR construct prepared using the LV-MAX™ Lentiviral Production System at a low multiplicity of infection (MOI) of 2.
Figure 2a: One-step isolation/activation of T cells from quarter Leukopaks, followed by lentivirus-based editing. The process is closed, and digital automation occurs through Cellmation software.
3. Debeading: Following the viral transduction, the cells were debeaded using the Gibco™ CTS™ Detachable Dynabeads™ Release Buffer on the CTS™ DynaCellect system. The active-release buffer enables users to detach the cells from the Dynabeads at any point post-isolation.
4. Wash and concentrate: The cells were then washed and concentrated using the Gibco CTS Rotea™ Counterflow Centrifugation System. The low-shear environment ensures minimal damage to the cells, high recovery and viability.
5. Cryopreservation and expansion: After the cells were washed and concentrated, they were divided into two parts. One part was cryopreserved using the Thermo Scientific CryoMed™ Controlled-Rate Freezer. At the same time, the other part was expanded for seven days for comparative studies.
Figure 2b: Debeading is done using the CTS Detachable Dynabeads Release Buffer with the CTS DynaCellect system.
The end-to-end process was digitally integrated and automated using the Gibco CTS Cellmation Software for DeltaV System. This workflow allowed for CAR-T cell production within 24 hours. The results from the comparative profiling indicate that the T cells exhibit a more naive memory/T stem cell memory (TSCM) phenotype 24 hours after transduction, whereas the cells cultured for seven days show a more differentiated phenotype (Figure 3).
Figure 3: Data showing a higher naive TSCM phenotype (CD45RA+/CCR7+) population CAR-T cells manufactured utilizing the 24-hour workflow, as compared to the 7-day process.
For detailed insights into the story, access our on-demand webinar here: Webinar: An Automated 24-hour CAR-T Manufacturing Process | Thermo Fisher Scientific – US
The future of cell therapies and decentralized models of manufacturing
The success and quality of autologous cell therapies are heavily reliant on the overall health of the patients from whom the cells are sourced, which is a significant limitation in their development. Additionally, complex protocols with long activation and expansion steps lead to long production timelines. Ultimately, this can affect the timely delivery of therapy to patients, potentially increase costs, and may impact therapeutic efficacy.
Cell therapies are a unique modality and lacked established frameworks in the early years of clinical manufacturing. Due to a lack of good precedents, most of the practices from traditional pharmacological manufacturing were adapted for autologous cell therapies, which extends timelines significantly while increasing the burden of financial reimbursements. Although manufacturing accounts for nearly 50% of the standard vein-to-vein timeline, prolonged cell expansion ex vivo can significantly impact the quality of the final products. Cell therapies have a different curative approach which relies on the desired mechanism of action (MOA) that the therapeutic cells exert.
Decentralized systems shift the processes of manufacturing and distribution to localized sites, making therapies readily accessible to patients. This requires fundamental changes in infrastructure, technology, business operations and regulatory landscapes (5, 9).
“Both of these models (centralized and decentralized) can and will coexist. This is an area that will generate a lot of excitement in terms of the future and some of these new approaches. When it comes to start-ups, biotechs, and academic centers, I do feel that the decentralized approach represents a novel manufacturing space where these players can now demonstrate their drugs.”
Fabio Fachin, Ph.D.,
Executive Director, Cell Therapy, Takeda
The advent of next-generation technologies, integration of instruments with digital automation and increased modularity of operations can help enable decentralized, point-of-care manufacturing for autologous cell therapies.
To learn more about the 24-hour autologous CAR-T cell manufacturing process, read the full story here: Shorten the CAR-T manufacturing process | Thermo Fisher Scientific – US
To learn more about cell and gene therapy instrumentation, visit our website: www.thermofisher.com/ctxmanufacturing
Request a demo: Gibco CTS DynaCellect Magnetic Separation System | Thermo Fisher Scientific – US
References:
- Kalos M, et al. T cells with chimeric antigen receptors have potent antitumor effects and can establish memory in patients with advanced leukemia. Sci Transl Med. 2011;3(95):95ra73.
- Tyagarajan S, et al. Optimizing CAR-T Cell Manufacturing Processes during Pivotal Clinical Trials. Mol Ther Methods Clin Dev. 2019; 16:136-144. Published 2019 Nov 29.
- Abou-El-Enein M, et al. Scalable Manufacturing of CAR T cells for Cancer Immunotherapy. Blood Cancer Discov. 2021;2(5):408-422.
- Rodrigues M, et al. Optimizing commercial manufacturing of tisagenlecleucel for patients in the US: a 4-year experiential journey. Blood 2021;138 Suppl 1:1768. 26.
- Harrison RP, et al. Decentralized manufacturing of cell and gene therapies: Overcoming challenges and identifying opportunities. Cytotherapy. 2017;19(10):1140-1151.
- Dickinson MJ, et al. A Novel Autologous CAR-T Therapy, YTB323, with Preserved T-cell Stemness Shows Enhanced CAR T-cell Efficacy in Preclinical and Early Clinical Development. Cancer Discov. 2023;13(9):1982-1997. doi:10.1158/2159-8290.CD-22-1276
- Ghassemi S, et al. Rapid manufacturing of non-activated potent CAR T cells. Nat Biomed Eng. 2022;6(2):118-128
- Yang J, et al. Next-day manufacture of a novel anti-CD19 CAR-T therapy for B-cell acute lymphoblastic leukemia: first-in-human clinical study. Blood Cancer J. 2022;12(7):104. Published 2022 Jul 7.
- Palani, H.K., et al. Decentralized manufacturing of anti CD19 CAR-T cells using CliniMACS Prodigy®: real-world experience and cost analysis in India. Bone Marrow Transplant 58, 160–167 (2023).
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