Critical considerations for the development of potency tests for therapeutic applications of mesenchymal stromal cell-derived small extracellular vesicles

Mario Gimona; Maria Felice Brizzi; Andre Boon Hwa Choo; Massimo Dominici; Sean M Davidson; Johannes Grillari; Dirk M Hermann; Andrew F Hill; Dominique de Kleijn; Ruenn Chai Lai; Charles P Lai; Rebecca Lim; Marta Monguió-Tortajada; Maurizio Muraca; Takahiro Ochiya; Luis A Ortiz; Wei Seong Toh; Yong Weon Yi; Kenneth W Witwer; Bernd Giebel; Sai Kiang Lim

doi:10.1016/j.jcyt.2021.01.001

Critical considerations for the development of potency tests for therapeutic applications of mesenchymal stromal cell-derived small extracellular vesicles

Cytotherapy. 2021 May;23(5):373-380. doi: 10.1016/j.jcyt.2021.01.001. Epub 2021 Apr 10.

Authors

Mario Gimona¹, Maria Felice Brizzi², Andre Boon Hwa Choo³, Massimo Dominici⁴, Sean M Davidson⁵, Johannes Grillari⁶, Dirk M Hermann⁷, Andrew F Hill⁸, Dominique de Kleijn⁹, Ruenn Chai Lai¹⁰, Charles P Lai¹¹, Rebecca Lim¹², Marta Monguió-Tortajada¹³, Maurizio Muraca¹⁴, Takahiro Ochiya¹⁵, Luis A Ortiz¹⁶, Wei Seong Toh¹⁷, Yong Weon Yi¹⁸, Kenneth W Witwer¹⁹, Bernd Giebel²⁰, Sai Kiang Lim²¹

Affiliations

¹ Good Manufacturing Practice Laboratory, Spinal Cord Injury and Tissue Regeneration Center Salzburg and Research Program Nanovesicular Therapies, Paracelsus Medical University, Salzburg, Austria.
² Department of Medical Sciences and Molecular Biotechnology Center, University of Torino, Torino, Italy.
³ Bioprocessing Technology Institute, Agency for Science, Technology and Research, Singapore, Singapore.
⁴ Technopole Mario Veronesi, Mirandola, Italy; Division of Medical Oncology, Laboratory of Cellular Therapy, University of Modena and Reggio Emilia, Modena, Italy.
⁵ The Hatter Cardiovascular Institute, University College London, London, UK.
⁶ Ludwig Boltzmann Institute for Experimental and Clinical Traumatology, Vienna, Austria; Department of Biotechnology, Christian Doppler Laboratory on Biotechnology of Skin Aging, Institute for Molecular Biotechnology, University of Natural Resources and Life Sciences, Vienna, Austria.
⁷ Department of Neurology, University Hospital Essen, University of Duisburg-Essen, Essen, Germany.
⁸ Department of Biochemistry and Genetics, La Trobe Institute for Molecular Science, La Trobe University, Bundoora, Australia.
⁹ Department of Vascular Surgery, University Medical Center Utrecht, Utrecht, the Netherlands.
¹⁰ Institute of Medical Biology and Institute of Molecular and Cell Biology, Agency for Science, Technology and Research, Singapore, Singapore.
¹¹ Institute of Atomic and Molecular Sciences, Academia Sinica, Taipei, Taiwan.
¹² Department of Obstetrics and Gynecology, The Ritchie Centre, Hudson Institute of Medical Research, Monash University, Victoria, Australia.
¹³ ICREC Research Program and REMAR-IVECAT group, Health Science Research Institute Germans Trias i Pujol (IGTP), Can Ruti Campus, and Cardiology Service, Germans Trias i Pujol University Hospital, Badalona, Spain.
¹⁴ Department of Women's and Children's Health, University of Padova, Padova, Italy.
¹⁵ Department of Molecular and Cellular Medicine, Tokyo Medical University, Tokyo, Japan.
¹⁶ Department of Environmental and Occupational Health, Division of Environmental and Occupational Medicine, Graduate School of Public Health at the University of Pittsburgh, Pittsburgh, Pennsylvania, USA.
¹⁷ Faculty of Dentistry, National University of Singapore, Singapore, Singapore; Department of Orthopaedic Surgery, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore.
¹⁸ ExoCoBio Exosome Institute, ExoCoBio Inc., Seoul, Korea.
¹⁹ Department of Molecular and Comparative Pathobiology, The Johns Hopkins University School of Medicine, Baltimore, USA; Department of Neurology, The Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.
²⁰ Institute for Transfusion Medicine, University Hospital Essen, University of Duisburg-Essen, Essen, Germany. Electronic address: [email protected].
²¹ Institute of Medical Biology and Institute of Molecular and Cell Biology, Agency for Science, Technology and Research, Singapore, Singapore; Department of Surgery, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore. Electronic address: [email protected].

PMID: 33934807
DOI: 10.1016/j.jcyt.2021.01.001

Abstract

Mesenchymal stromal/stem cells (MSCs) have been widely tested against many diseases, with more than 1000 registered clinical trials worldwide. Despite many setbacks, MSCs have been approved for the treatment of graft-versus-host disease and Crohn disease. However, it is increasingly clear that MSCs exert their therapeutic functions in a paracrine manner through the secretion of small extracellular vesicles (sEVs) of 50-200 nm in diameter. Unlike living cells that can persist long-term, sEVs are non-living and non-replicative and have a transient presence in the body. Their small size also renders sEV preparations highly amenable to sterilization by filtration. Together, acellular MSC-sEV preparations are potentially safer and easier to translate into the clinic than cellular MSC products. Nevertheless, there are inherent challenges in the development of MSC-sEV drug products. MSC-sEVs are products of living cells, and living cells are sensitive to changes in the external microenvironment. Consequently, quality control metrics to measure key identity and potency features of MSC-sEV preparations have to be specified during development of MSC-sEV therapeutics. The authors have previously described quantifiable assays to define the identity of MSC-sEVs. Here the authors discuss requirements for prospective potency assays to predict the therapeutic effectiveness of the drug substance in accordance with International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use guidelines. Although potency assays should ideally reflect the mechanism of action (MoA), this is challenging because the MoA for the reported efficacy of MSC-sEV preparations against multiple diseases of diverse underlying pathology is likely to be complex and different for each disease and difficult to fully elucidate. Nevertheless, robust potency assays could be developed by identifying the EV attribute most relevant to the intended biological activity in EV-mediated therapy and quantifying the EV attribute. Specifically, the authors highlight challenges and mitigation measures to enhance the manufacture of consistent and reproducibly potent sEV preparations, to identify and select the appropriate EV attribute for potency assays despite a complex "work-in-progress" MoA and to develop assays likely to be compliant with regulatory guidance for assay validation.

Keywords: MSC-EVs intervention; exosomes; extracellular vesicles; mesenchymal stromal cells; mesencyhmal stem cells; microvesicles.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Extracellular Vesicles*
Graft vs Host Disease*
Humans
Mesenchymal Stem Cells*
Prospective Studies

Abstract

Publication types

MeSH terms

Grants and funding