title: Controlled aggregation of ferritin to modulate MRI relaxivity.
abstract: Ferritin is an iron storage protein expressed in varying concentrations in mammalian cells. The deposition of ferric iron in the core of ferritin makes it a magnetic resonance imaging contrast agent, and ferritin has recently been proposed as a gene expression reporter protein for magnetic resonance imaging. To date, ferritin has been overexpressed in vivo and has been coexpressed with transferrin receptor to increase iron loading in cells. However, ferritin has a relatively low T(2) relaxivity (R(2) approximately 1 mM(-1)s(-1)) at typical magnetic field strengths and so requires high levels of expression to be detected. One way to modulate the transverse relaxivity of a superparamagnetic agent is to cause it to aggregate, thereby manipulating the magnetic field gradients through which water diffuses. In this work, it is demonstrated by computer simulation and in vitro that aggregation of ferritin can alter relaxivity. The effects of aggregate size and intraaggregate perturber spacing on R(2) are studied. Computer modeling indicates that the optimal spacing of the ferritin molecules in aggregate for increasing R(2) is 100-200 nm for a typical range of water diffusion rates. Chemical cross-linking of ferritin at 12 A spacing led to a 70% increase in R(2) compared to uncross-linked ferritin controls. To modulate ferritin aggregation in a potentially biologically relevant manner, ferritin was attached to actin and polymerized in vitro. The polymerization of ferritin-F-actin caused a 20% increase in R(2) compared to unpolymerized ferritin-G-actin. The R(2)-value was increased by another 10% by spacing the ferritin farther apart on the actin filaments. The modulation of ferritin aggregation by binding to cytoskeletal elements may be a useful strategy to make a functional reporter gene for magnetic resonance imaging.
authors.affiliation: Laboratory of Functional and Molecular Imaging, National Institutes of Neurological Disease and Stroke, National Institutes of Health, Bethesda, MD 20892-1065, USA.
authors.forename: Kevin M
authors.initials: KM
authors.lastname: Bennett
authors.forename: Erik M
authors.initials: EM
authors.lastname: Shapiro
authors.forename: Christopher H
authors.initials: CH
authors.lastname: Sotak
authors.forename: Alan P
authors.initials: AP
authors.lastname: Koretsky
mesh_terms: D003287:Contrast Media / Q000737:chemistry*; D005293:Ferritins / Q000737:chemistry*; D017930:Genes, Reporter; D007089:Image Enhancement / Q000379:methods*; D008279:Magnetic Resonance Imaging / Q000295:instrumentation / Q000379:methods*; D019047:Phantoms, Imaging; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D003287:Contrast Media; D005293:Ferritins