Modular Pore-Forming Immunotoxins with Caged Cytotoxicity Tailored by Directed Evolution.
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Authors
Soskine, Misha
Huang, Gang
Albuquerque, Inês S
Publication Date
2018-11-16Journal Title
ACS Chem Biol
ISSN
1554-8929
Publisher
American Chemical Society (ACS)
Volume
13
Issue
11
Pages
3153-3160
Language
eng
Type
Article
Physical Medium
Print-Electronic
Metadata
Show full item recordCitation
Mutter, N. L., Soskine, M., Huang, G., Albuquerque, I. S., Bernardes, G. J., & Maglia, G. (2018). Modular Pore-Forming Immunotoxins with Caged Cytotoxicity Tailored by Directed Evolution.. ACS Chem Biol, 13 (11), 3153-3160. https://doi.org/10.1021/acschembio.8b00720
Abstract
Immunotoxins are proteins containing a cell-targeting element linked to a toxin that are under investigation for next-generation cancer treatment. However, these agents are difficult to synthesize, chemically heterogeneous, expensive, and show toxicity toward healthy cells. In this work, we describe the synthesis and characterization of a new type of immunotoxin that showed exquisite selectivity toward targeted cells. In our construct, targeting molecules were covalently attached or genetically fused to oligomeric pore-forming toxins. The activity of the immunotoxin was then caged by fusing a soluble protein to the transmembrane domain and activated via cleavage with furin, which is a protease that is overexpressed in many cancer cells. During the several coupling steps, directed evolution allowed the efficient synthesis of the molecules in E. coli cells, as well as selection for further specificity toward targeted cells. The final construct showed no off-target activity, while acquiring an additional degree of specificity toward the targeted cells upon activation. The pore-forming toxins described here do not require internalization to operate, while the many protomeric subunits can be individually modified to refine target specificity.
Keywords
Animals, Cell Line, Tumor, Cell Membrane, Cnidarian Venoms, Directed Molecular Evolution, Drug Design, Escherichia coli, Folic Acid, Furin, Humans, Immunotoxins, Mutagenesis, Pore Forming Cytotoxic Proteins, Proteolysis, Recombinant Fusion Proteins, Salmonella typhi, Sea Anemones, Tetrahydrofolate Dehydrogenase
Sponsorship
The Royal Society (uf110046)
European Research Council (676832)
Identifiers
External DOI: https://doi.org/10.1021/acschembio.8b00720
This record's URL: https://www.repository.cam.ac.uk/handle/1810/286307
Rights
Attribution-NonCommercial-NoDerivatives 4.0 International
Licence URL: https://creativecommons.org/licenses/by-nc-nd/4.0/
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