Industrializing Autologous Adoptive Immunotherapies: Manufacturing Advances and Challenges

Abstract

Cell therapy has proven to be a burgeoning field of investigation, evidenced by hundreds of clinical trials being conducted worldwide across a variety of cell types and indications. Many cell therapies have been shown to be efficacious in humans, such as modified T-cells and natural killer (NK) cells. Adoptive immunotherapy has shown the most promise in recent years, with particular emphasis on autologous cell sources. Chimeric Antigen Receptor (CAR)-based T-cell therapy targeting CD19-expressing B-cell leukemias has shown remarkable efficacy and reproducibility in numerous clinical trials. Recent marketing approval of Novartis' Kymriah™ (tisagenlecleucel) and Gilead/Kite's Yescarta™ (axicabtagene ciloleucel) by the FDA further underscores both the promise and legwork to be done if manufacturing processes are to become widely accessible. Further work is needed to standardize, automate, close, and scale production to bring down costs and democratize these and other cell therapies. Given the multiple processing steps involved, commercial-scale manufacturing of these therapies necessitates tighter control over process parameters. This focused review highlights some of the most recent advances used in the manufacturing of therapeutic immune cells, with a focus on T-cells. We summarize key unit operations and pain points around current manufacturing solutions. We also review emerging technologies, approaches and reagents used in cell isolation, activation, transduction, expansion, in-process analytics, harvest, cryopreservation and thaw, and conclude with a forward-look at future directions in the manufacture of adoptive immunotherapies.

Targeting PVR (CD155) and its receptors in anti-tumor therapy

“…The expression of these receptors confers potent effector capacity to these cells, ultimately leading to their robust activation and killing of the target tumor cells. [156][157][158] Usually, CARs are engineered to contain an extracellular antigen-recognizing region usually antibody-derived single-chain variable fragment linked to an intracellular signaling domain, usually TCR/CD3 ζ-chain and co-stimulatory domains. 156 Since antigen recognition of these cells does not depend on MHC, CAR T cells are able to circumvent some of the MHC-targeted tumor immune evasion mechanisms.…”

Automation: what will the cell therapy laboratory of the future look like?

“…Novel technologies for cell separations using microbubbles and sonic waves are under development and have shown promising results [28]. Expected improvements in antibody-based separations will increase sensitivity and reduce both cost and processing time.…”

“…Even di-methyl sulfoxide (DMSO) could be eliminated from the lab of the future. Some tubing sets (such as those made with polyvinyl chloride [PVC]) are not compatible with sterile welding in the presence of DMSO and prolonged exposure to DMSO may be cytotoxic [28].…”

“…Several controlled thaw devices are available. Asymptote VIA Thaw SC2 (Asymptote/ GE) and BioCision ThawSTAR Automated Cell Thawing System (BioCision) are designed to dry thaw vials while the Asymptote VIA Thaw CB1000, Medcision ThawCB and the Sarstedt SAHA-RA system other are capable of thawing bags [28].…”

Nanoparticle‐Mediated Intracellular Protection of Natural Killer Cells Avoids Cryoinjury and Retains Potent Antitumor Functions

“…[ 2 ] Dimethylsulfoxide (DMSO), the most commonly used cryoprotectant, is widely ubiquitous largely due to its efficiency and lack of equivalent alternative. [ 3 ] However, it has been associated with severe toxicities when infused into patients. [ 4–13 ] Adverse effects of DMSO have also been studied extensively in vitro, with studies showing DMSO to be toxic to various cell types including blood cells, [ 14,15 ] mesenchymal stromal cells, [ 16 ] skin fibroblasts, [ 17,18 ] and human corneal endothelial cells.…”