title: Functionalized Biopolymer Particles Enhance Performance of a Tissue-Protective Peptide under Proteolytic and Thermal Stress.
abstract: Cutaneous burns are often exacerbated by poor perfusion and subsequent necrosis of the microvasculature surrounding the primary injury. Preservation of these vessels can reduce necrotic tissue expansion and increase success rates of skin graft procedures. Recent work has identified a peptide derived from erythropoietin, ARA290, with the ability to mediate tissue protection in a variety of cell types. Here we demonstrate the advantages of fusing ARA290 to an elastin-like polypeptide (ELP) to salvage microvascular endothelial cells in harsh proteolytic conditions following thermal shock. These fusion proteins were expressed recombinantly in bacterial hosts and rapidly purified by inverse transition cycling. They were shown to spontaneously aggregate into particles at subphysiological temperatures. The bifunctional submicron particles were resistant to digestion in enzymes upregulated after burn injury. Furthermore, the data strongly suggest these ARA290-functionalized particles were superior to treatment with the peptide alone in preventing microvascular cell death in these conditions. The results bring to light an efficient and cost-effective strategy for the delivery therapeutic peptides to proteolytically active wound sites.
authors.affiliation: Center for Engineering in Medicine, Massachusetts General Hospital, Harvard Medical School, Shriners Hospitals for Children, Boston, Massachusetts 02114, United States.
authors.forename: Kevin
authors.initials: K
authors.lastname: Dooley
authors.affiliation: Center for Engineering in Medicine, Massachusetts General Hospital, Harvard Medical School, Shriners Hospitals for Children, Boston, Massachusetts 02114, United States.
authors.forename: Julie
authors.initials: J
authors.lastname: Devalliere
authors.affiliation: Center for Engineering in Medicine, Massachusetts General Hospital, Harvard Medical School, Shriners Hospitals for Children, Boston, Massachusetts 02114, United States.
authors.forename: Basak E
authors.initials: BE
authors.lastname: Uygun
authors.affiliation: Center for Engineering in Medicine, Massachusetts General Hospital, Harvard Medical School, Shriners Hospitals for Children, Boston, Massachusetts 02114, United States.|Department of Biomedical Engineering, Rutgers University , Piscataway, New Jersey 08854, United States.
authors.forename: Martin L
authors.initials: ML
authors.lastname: Yarmush
mesh_terms: D001704:Biopolymers / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology*; D002056:Burns / Q000517:prevention & control; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D004549:Elastin / Q000737:chemistry* / Q000235:genetics / Q000494:pharmacology; D004921:Erythropoietin / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics; D006358:Hot Temperature; D006801:Humans; D055806:Microvessels / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D009842:Oligopeptides / Q000737:chemistry* / Q000235:genetics / Q000494:pharmacology; D059748:Proteolysis / Q000187:drug effects*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology*; D012867:Skin / Q000098:blood supply; D014945:Wound Healing / Q000187:drug effects
chemical_list: D001704:Biopolymers; D009842:Oligopeptides; D011993:Recombinant Fusion Proteins; D004921:Erythropoietin; D004549:Elastin; C576178:cibinetide