title: Targeted magnetic resonance imaging of Scavidin-receptor in human umbilical vein endothelial cells in vitro.
abstract: Current therapeutic approaches to treat cancer are often hampered by the lack of specificity of the drugs used for therapy. Scavidin, a novel fusion protein expressed on cell membranes, could be utilized for targeting of therapeutic molecules. Scavidin exploits the high binding affinity between avidin and biotin and is capable of mediating endocytosis of a bound ligand. In the current study we evaluated the efficiency of biotinylated ultrasmall superparamagnetic iron oxide (USPIO) particles in Scavidin-expressing human umbilical vein endothelial cell (HUVEC) cultures in vitro as a novel receptor-targeted magnetic resonance imaging contrast agent. Biotinylated USPIO (bUSPIO) were targeted to Scavidin adenovirus-transduced HUVECs in vitro. Scavidin expressing cells were capable of binding and mediating endocytosis of the bUSPIO in vitro, which led to a significant decrease in T2 relaxation times, and a loss of signal intensity in comparison to controls. The findings were confirmed with Prussian blue staining for iron and detection of Scavidin by bound biotinylated horseradish peroxidase. Our data shows that biotinylated ligands target specifically to Scavidin-expressing HUVEC in vitro. The utilization of Scavidin gene transfer ex vivo thus constitutes a platform for potential ligand delivery via cell therapy and time-independent imaging of biologic processes.
authors.affiliation: A.I. Virtanen Institute, University of Kuopio, Kuopio, Finland.
authors.forename: Tuomas
authors.initials: T
authors.lastname: Mäntylä
authors.forename: Juhana M
authors.initials: JM
authors.lastname: Hakumäki
authors.forename: Tuulia
authors.initials: T
authors.lastname: Huhtala
authors.forename: Ale
authors.initials: A
authors.lastname: Närvänen
authors.forename: Seppo
authors.initials: S
authors.lastname: Ylä-Herttuala
mesh_terms: D000256:Adenoviridae; D015153:Blotting, Western; D002462:Cell Membrane / Q000378:metabolism*; D002478:Cells, Cultured; D003001:Cloning, Molecular; D003911:Dextrans; D052203:Ferrosoferric Oxide; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors; D006801:Humans; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D007501:Iron; D008279:Magnetic Resonance Imaging / Q000379:methods*; D058185:Magnetite Nanoparticles; D010087:Oxides; D011971:Receptors, Immunologic / Q000737:chemistry* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014471:Umbilical Veins / Q000166:cytology*
chemical_list: D003911:Dextrans; D058185:Magnetite Nanoparticles; D010087:Oxides; D011971:Receptors, Immunologic; D011993:Recombinant Fusion Proteins; C097921:ferumoxtran-10; C451404:scavidin; D007501:Iron; D052203:Ferrosoferric Oxide