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 α(2)-plasmin inhibitor (α(2)PI(1-8)), the resulting variant, α(2)PI(1-8)-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 α(2)PI(1-8)-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 α(2)PI(1-8)-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.
Institute of Bioengineering, Ecole Polytechnique Fédérale de Lausanne, CH-1015 Lausanne, Switzerland.
Kristen M
KM
Lorentz
Lirong
L
Yang
Peter
P
Frey
Jeffrey A
JA
Hubbell
D000818:Animals; D049109:Cell Proliferation; D002478:Cells, Cultured; D020224:Expressed Sequence Tags; D005260:Female; D005337:Fibrin / Q000378:metabolism; D005818:Genetic Engineering; D006801:Humans; D007334:Insulin-Like Growth Factor I / Q000235:genetics / Q000378:metabolism*; D009130:Muscle, Smooth / Q000502:physiology*; D032389:Myocytes, Smooth Muscle / Q000378:metabolism; D010766:Phosphorylation; D051381:Rats; D017207:Rats, Sprague-Dawley; D017526:Receptor, IGF Type 1 / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012038:Regeneration; D001743:Urinary Bladder / Q000166:cytology / Q000378:metabolism; D000979:alpha-2-Antiplasmin / Q000378:metabolism
D011994:Recombinant Proteins; D000979:alpha-2-Antiplasmin; D007334:Insulin-Like Growth Factor I; D005337:Fibrin; D017526:Receptor, IGF Type 1