title: Directed evolution of a soluble human DR3 receptor for the inhibition of TL1A induced cytokine secretion.
abstract: TNF-like 1A (TL1A) is a cytokine belonging to the TNF superfamily that promotes inflammation in autoimmune diseases. Inhibiting the interaction of TL1A with the endogenous death-domain receptor 3 (DR3) offers a therapeutic approach for treating TL1A-induced autoimmune diseases. Here, we generated improved DR3 variants showing increased TL1A binding affinity and stability using a directed evolution approach. Given the high cysteine content and post-translational modification of DR3, we employed yeast surface display and expression in mammalian cell lines for screening, expression and characterization of improved DR3 variants. A cell-based assay performed with the human TF-1 cell line and CD4+ T cells showed that two improved DR3 mutants efficiently inhibited TL1A-induced cell death and secretion of IFN-γ, respectively. These DR3 mutants can be used as drug candidates for the treatment of inflammatory bowel diseases and for other autoimmune diseases, including rheumatic arthritis and asthma.
authors.affiliation: The National Institute for Biotechnology in the Negev (NIBN), Ben-Gurion University of the Negev, Be'er Sheva, Israel.
authors.forename: Itay
authors.initials: I
authors.lastname: Levin
authors.affiliation: The National Institute for Biotechnology in the Negev (NIBN), Ben-Gurion University of the Negev, Be'er Sheva, Israel.|Department of Life Sciences, Ben-Gurion University of the Negev, Be'er Sheva, Israel.
authors.forename: Marianna
authors.initials: M
authors.lastname: Zaretsky
authors.affiliation: The National Institute for Biotechnology in the Negev (NIBN), Ben-Gurion University of the Negev, Be'er Sheva, Israel.|Department of Life Sciences, Ben-Gurion University of the Negev, Be'er Sheva, Israel.
authors.forename: Amir
authors.initials: A
authors.lastname: Aharoni
mesh_terms: D017209:Apoptosis / Q000235:genetics; D002460:Cell Line; D019020:Directed Molecular Evolution; D057809:HEK293 Cells; D006801:Humans; D009154:Mutation; D053219:Receptors, Tumor Necrosis Factor, Member 25 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D012995:Solubility; D013601:T-Lymphocytes / Q000166:cytology / Q000378:metabolism; D053380:Tumor Necrosis Factor Ligand Superfamily Member 15 / Q000378:metabolism*
chemical_list: D053219:Receptors, Tumor Necrosis Factor, Member 25; C117026:TNFSF15 protein, human; D053380:Tumor Necrosis Factor Ligand Superfamily Member 15