title: New clathrin-based nanoplatforms for magnetic resonance imaging.
abstract: BACKGROUND
  Magnetic Resonance Imaging (MRI) has high spatial resolution, but low sensitivity for visualization of molecular targets in the central nervous system (CNS). Our goal was to develop a new MRI method with the potential for non-invasive molecular brain imaging. We herein introduce new bio-nanotechnology approaches for designing CNS contrast media based on the ubiquitous clathrin cell protein.
  
  
  RESULTS
  The first approach utilizes three-legged clathrin triskelia modified to carry 81 gadolinium chelates. The second approach uses clathrin cages self-assembled from triskelia and designed to carry 432 gadolinium chelates. Clathrin triskelia and cages were characterized by size, structure, protein concentration, and chelate and gadolinium contents. Relaxivity was evaluated at 0.47 T. A series of studies were conducted to ascertain whether fluorescent-tagged clathrin nanoplatforms could cross the blood brain barriers (BBB) unaided following intranasal, intravenous, and intraperitoneal routes of administration. Clathrin nanoparticles can be constituted as triskelia (18.5 nm in size), and as cages assembled from them (55 nm). The mean chelate: clathrin heavy chain molar ratio was 27.04±4.8: 1 for triskelia, and 4.2±1.04: 1 for cages. Triskelia had ionic relaxivity of 16 mM(-1) s(-1), and molecular relaxivity of 1,166 mM(-1) s(-1), while cages had ionic relaxivity of 81 mM(-1) s(-1) and molecular relaxivity of 31,512 mM(-1) s(-1). Thus, cages exhibited 20 times higher ionic relaxivity and 8,000-fold greater molecular relaxivity than gadopentetate dimeglumine. Clathrin nanoplatforms modified with fluorescent tags were able to cross or bypass the BBB without enhancements following intravenous, intraperitoneal and intranasal administration in rats.
  
  
  CONCLUSIONS
  Use of clathrin triskelia and cages as carriers of CNS contrast media represents a new approach. This new biocompatible protein-based nanotechnology demonstrated suitable physicochemical properties to warrant further in vivo imaging and drug delivery studies. Significantly, both nanotransporters crossed and/or bypassed the BBB without enhancers. Thus, clathrin nanoplatforms could be an appealing alternative to existing CNS bio-nanotechnologies.
authors.affiliation: Laboratory of Developmental Psychopharmacology, Brain Imaging Center, Department of Psychiatry, Harvard Medical School, McLean Hospital, Belmont, Massachusetts, United States of America. gvitaliano@mclean.harvard.edu
authors.forename: Gordana D
authors.initials: GD
authors.lastname: Vitaliano
authors.forename: Franco
authors.initials: F
authors.lastname: Vitaliano
authors.forename: Jose D
authors.initials: JD
authors.lastname: Rios
authors.forename: Perry F
authors.initials: PF
authors.lastname: Renshaw
authors.forename: Martin H
authors.initials: MH
authors.lastname: Teicher
mesh_terms: D000818:Animals; D001812:Blood-Brain Barrier / Q000000981:diagnostic imaging / Q000378:metabolism; D001921:Brain / Q000098:blood supply / Q000000981:diagnostic imaging* / Q000378:metabolism; D002966:Clathrin / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure; D033922:Clathrin Heavy Chains / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure; D004591:Electrophoresis, Polyacrylamide Gel; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry; D007274:Injections, Intraperitoneal; D007275:Injections, Intravenous; D017879:Isothiocyanates / Q000737:chemistry; D008279:Magnetic Resonance Imaging / Q000379:methods*; D008297:Male; D046529:Microscopy, Electron, Transmission; D008958:Models, Molecular; D053758:Nanoparticles / Q000008:administration & dosage / Q000737:chemistry / Q000648:ultrastructure; D036103:Nanotechnology / Q000379:methods*; D004369:Pentetic Acid / Q000031:analogs & derivatives / Q000737:chemistry; D011487:Protein Conformation; D055503:Protein Multimerization; D011859:Radiography; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D002966:Clathrin; D017879:Isothiocyanates; C485704:isothiocyanate-benzyl-DTPA; D033922:Clathrin Heavy Chains; D004369:Pentetic Acid; D016650:Fluorescein-5-isothiocyanate