Engineering and Therapeutic Application of Single-Chain Bivalent TGF-β Family Traps
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Abstract
Deregulation of TGF-β superfamily signaling is a causative factor in many diseases. Here we describe a protein engineering strategy for the generation of single-chain bivalent receptor traps for TGF-β superfamily ligands. Traps were assembled using the intrinsically disordered regions flanking the structured binding domain of each receptor as "native linkers" between two binding domains. This yields traps that are approximately threefold smaller than antibodies and consists entirely of native receptor sequences. Two TGF-β type II receptor-based, single-chain traps were designed, termed (TβRII)2 and (TβRIIb)2, that have native linker lengths of 35 and 60 amino acids, respectively. Both single-chain traps exhibit a 100 to 1,000 fold higher in vitro ligand binding and neutralization activity compared with the monovalent ectodomain (TβRII-ED), and a similar or slightly better potency than pan–TGF-β–neutralizing antibody 1D11 or an Fc-fused receptor trap (TβRII-Fc). Despite its short in vivo half-life (<1 hour), which is primarily due to kidney clearance, daily injections of the (TβRII)2 trap reduced the growth of 4T1 tumors in BALB/c mice by 50%, an efficacy that is comparable with 1D11 (dosed thrice weekly). In addition, (TβRII)2 treatment of mice with established 4T1 tumors (100 mm3) significantly inhibited further tumor growth, whereas the 1D11 antibody did not. Overall, our results indicate that our rationally designed bivalent, single-chain traps have promising therapeutic potential. Mol Cancer Ther; 11(7); 1477–87. ©2012 AACR.
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Antibodies against integrins β1, β6 and β8 selectively impair TGF-β1 activation.
“Antibodies against integrins β1, β6 and β8 were shown to selectively impair TGF-β1 activation85,313–315”
From TGFβ biology in cancer progression and immunotherapy (2021) · cites this paper as [313]
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TGFβ biology in cancer progression and immunotherapy
Derynck, R., Turley, S. J., & Akhurst, R. J. (2021) · Nature Reviews Clinical Oncology · doi:10.1038/s41571-020-0403-1 · Cited by 944 · see citing works
- Authors
- Zwaagstra, J. C., Sulea, T., Baardsnes, J., Lenferink, A. E. G., Collins, C., Cantin, C., Paul-Roc, B., Grothe, S., Hossain, S., Richer, L.-P., L'Abbé, D., Tom, R., Cass, B., Durocher, Y., & O'Connor-McCourt, M. D.
- Affiliations
- Biotechnology Research Institute (CA)
- Authors' Affiliation: Biotechnology Research Institute, National Research Council Canada, Montreal, Quebec, Canada
- Corresponding author
- Zwaagstra, John C.
- ORCID
- Venue
- Molecular Cancer Therapeutics
- Published
- 2012-07-01
- Volume
- 11
- Issue
- 7
- Pages
- 1477-1487
- ISSN
- 1535-7163 (ISSN-L)
- 1538-8514
- OpenAlex
- W2140081727
- Cited by
- 20
- References
- 50
- Citations by year
- 2025: 1 · 2024: 3 · 2023: 1 · 2022: 1 · 2021: 5 · 2020: 2 · 2018: 1 · 2017: 1 · 2016: 3 · 2015: 2
- FWCI
- 0.26
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Molecular Cancer Therapeutics (publishedVersion)
- PubMed (publishedVersion)
- NPARC (submittedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › TGF-β signaling in diseases
- Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Monoclonal and Polyclonal Antibodies Research
- Life Sciences › Immunology and Microbiology › Immunology › Toxin Mechanisms and Immunotoxins
- Keywords
- TGF-beta superfamily, TGFBR2, intrinsically disordered regions, protein engineering, TGF-beta neutralization, 4T1 breast cancer, tumor growth inhibition, antibody alternatives, renal clearance
- MeSH terms
- Animals, Female, Humans, Immunosuppression Therapy, Ligands, Male, Mice, Inbred BALB C, Models, Molecular, Protein Binding, Protein Conformation, Recombinant Fusion Proteins, Protein Engineering, Transforming Growth Factor beta, Rats, Sprague-Dawley, Receptors, Transforming Growth Factor beta, Gene Order, Cell Line, Tumor, Cell Proliferation, Mice, Rats, Protein Stability
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