Engineered insulin-like growth factor-1 for improved smooth muscle regeneration

Insulin-like growth factor-1 (IGF-1) has been shown to induce potent mitogenic responses in various cell types, yet its sustained local delivery is still an underdeveloped domain in the clinic. We report here an engineered IGF-1 that facilitates extended local delivery to a site through its immobilization capacity within fibrin. Through recombinant fusion with a substrate sequence tag derived from α2-plasmin inhibitor (α2PI1-8), the resulting variant, α2PI1-8–IGF-1, was covalently incorporated into fibrin matrices during normal thrombin/factor XIIIa-mediated polymerization. Bioactivity of the variant was confirmed to be equivalent to wild type (WT) IGF-1 via IGF-1 receptor phosphorylation and cell proliferation studies in urinary tract-derived cells in 2-D. Assessment of functional retention within 3-D fibrin matrices demonstrated that incorporation of α2PI1–8–IGF-1 induced a 1.3- and 1.5-fold more robust proliferative response in smooth muscle cells (SMCs) than WT IGF-1 and negative control matrices, respectively, when release was not contained. Sustained α2PI1–8–IGF-1 availability at bladder lesion sites in vivo evoked a considerable increase in SMC proliferation and a favorable host tissue response after 28 days in rats. We conclude that the sustained local IGF-1 availability from fibrin provided by our variant protein enhances smooth muscle regeneration better than the WT form of the protein.