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OBJECTIVES: Targeting cytokines to sites of disease has clear advantages because it increases their therapeutic index. We designed fusion proteins of the latent-associated peptide (LAP) derived from TGF-β with various cytokines via a matrix metalloproteinase (MMP) cleavage site. This design confers latency, increased half-life and targeting to sites of inflammation. The aim of this study is to determine whether this approach can be applied to cytokines of different molecular structures and sizes. METHODS: Mature cytokines cloned downstream of LAP and a MMP cleavage site were expressed in 293T cells and assessed for latency and biological activity by Western blotting and bioassay. RESULTS: We demonstrate here that fusion proteins of TGF-β, erythropoietin, IL-1ra, IL-10, IL-4, BMP-7, IGF1 and IL-17 were rendered latent by fusion to LAP, requiring cleavage to become active in respective bioassays. As further proof of principle, we also show that delivery of engineered TGF-β can inhibit experimental autoimmune encephalomyelitis and that this approach can be used to efficiently deliver cytokines to the brain and spinal cord in mice with this disease. CONCLUSIONS: The latent cytokine approach can be successfully applied to a range of molecules, including cytokines of different molecular structure and mass, growth factors and a cytokine antagonist.

Original publication

DOI

10.1517/17425247.2013.839655

Type

Journal article

Journal

Expert Opin Drug Deliv

Publication Date

01/2014

Volume

11

Pages

5 - 16

Keywords

Animals, Cell Line, Cells, Cultured, Chick Embryo, Cytokines, Encephalomyelitis, Autoimmune, Experimental, Fibroblasts, HEK293 Cells, HeLa Cells, Humans, Insulin-Like Growth Factor I, Matrix Metalloproteinase 1, Mice, Mice, Inbred DBA, Mink, Molecular Targeted Therapy, Peptides, Protein Precursors, Recombinant Fusion Proteins, Transforming Growth Factor beta