Mutants of lymphotoxin-α with augmented cytotoxic activity via TNFR1 for use in cancer therapy.
Cytokine
; 61(2): 578-84, 2013 Feb.
Article
em En
| MEDLINE
| ID: mdl-23246116
ABSTRACT
The cytokine lymphotoxin-α (LTα) is a promising candidate for use in cancer therapy. However, the instability of LTαin vivo and the insufficient levels of tumor necrosis factor receptor 1 (TNFR1)-mediated bioactivity of LTα limit its therapeutic potential. Here, we created LTα mutants with increased TNFR1-mediated bioactivity by using a phage display technique. We constructed a phage library displaying lysine-deficient structural variants of LTα with randomized amino acid residues. After affinity panning, we screened three clones of lysine-deficient LTα mutant, and identified a LTα mutant with TNFR1-mediated bioactivity that was 32 times that of the wild-type LTα (wtLTα). When compared with wtLTα, the selected clone showed augmented affinity to TNFR1 due to slow dissociation rather than rapid association. In contrast, the mutant showed only 4 times the TNFR2-mediated activity of wtLTα. In addition, the LTα mutant strongly and rapidly activated caspases that induce TNFR1-mediated cell death, whereas the mutant and wtLTα activated nuclear factor-kappa B to a similar extent. Our data suggest that the kinetics of LTα binding to TNFR1 play an important role in signal transduction patterns, and a TNFR1-selective LTα mutant with augmented bioactivity would be a superior candidate for cancer therapy.
Texto completo:
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Base de dados:
MEDLINE
Assunto principal:
Linfotoxina-alfa
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Receptores Tipo I de Fatores de Necrose Tumoral
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Proteínas Mutantes
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Neoplasias
Idioma:
En
Ano de publicação:
2013
Tipo de documento:
Article