A clinically relevant large-scale biomanufacturing workflow to produce natural killer cells and natural killer cell-derived extracellular vesicles for cancer immunotherapy

Frederic St-Denis-Bissonnette; Sarah E Cummings; Shirley Qiu; Andrew Stalker; Gauri Muradia; Jelica Mehic; Karan Mediratta; Shelby Kaczmarek; Dylan Burger; Seung-Hwan Lee; Lisheng Wang; Jessie R Lavoie

doi:10.1002/jev2.12387

A clinically relevant large-scale biomanufacturing workflow to produce natural killer cells and natural killer cell-derived extracellular vesicles for cancer immunotherapy

J Extracell Vesicles. 2023 Dec;12(12):e12387. doi: 10.1002/jev2.12387.

Authors

Frederic St-Denis-Bissonnette^{1

2}, Sarah E Cummings¹, Shirley Qiu¹, Andrew Stalker¹, Gauri Muradia¹, Jelica Mehic¹, Karan Mediratta^{2

3

4}, Shelby Kaczmarek^{2

3}, Dylan Burger^{5

6}, Seung-Hwan Lee^{2

3}, Lisheng Wang^{2

3

4

7}, Jessie R Lavoie^{1

2}

Affiliations

¹ Centre for Oncology, Radiopharmaceuticals and Research, Biologic and Radiopharmaceutical Drugs Directorate, Health Products and Food Branch, Health Canada, Ottawa, ON, Canada.
² Department of Biochemistry, Microbiology and Immunology, Faculty of Medicine, University of Ottawa, Ottawa, ON, Canada.
³ Centre for Infection, Immunity and Inflammation, University of Ottawa, Ottawa, ON, Canada.
⁴ Ottawa Institute of Systems Biology, University of Ottawa, Ottawa, ON, Canada.
⁵ Kidney Research Centre, Ottawa Hospital Research Institute, Ottawa, ON, Canada.
⁶ Department of Cellular and Molecular Medicine, University of Ottawa, Ottawa, ON, Canada.
⁷ Regenerative Medicine Program, Ottawa Hospital Research Institute, Ottawa, ON, Canada.

Abstract

Natural killer cell-derived extracellular vesicles (NK-EVs) have shown promising potential as biotherapeutics for cancer due to their unique attributes as cytotoxic nanovesicles against cancer cells and immune-modulatory activity towards immune cells. However, a biomanufacturing workflow is needed to produce clinical-grade NK-EVs for pre-clinical and clinical applications. This study established a novel biomanufacturing workflow using a closed-loop hollow-fibre bioreactor to continuously produce NK-EVs from the clinically relevant NK92-MI cell line under serum-free, Xeno-free and feeder-free conditions following GMP-compliant conditions. The NK92 cells grown in the bioreactor for three continuous production lots resulted in large quantities of both NK cell and NK-EV biotherapeutics at the end of each production lot (over 10⁹ viable cells and 10¹³ EVs), while retaining their cytotoxic payload (granzyme B and perforin), pro-inflammatory cytokine (interferon-gamma) content and cytotoxicity against the human leukemic cell line K562 with limited off-target toxicity against healthy human fibroblast cells. This scalable biomanufacturing workflow has the potential to facilitate the clinical translation of adoptive NK cell-based and NK-EV-based immunotherapies for cancer with GMP considerations.

Keywords: biomanufacturing workflow; cancer immunotherapy; clinical-grade; extracellular vesicles (EVs); natural killer (NK) cells; scalable production.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Extracellular Vesicles*
Humans
Immunotherapy / methods
Killer Cells, Natural
Neoplasms* / metabolism
Neoplasms* / therapy
Workflow

Grants and funding

Operating Grant 175177/CIHR/Canada