Molecular imaging of heparan sulfate expression with radiolabeled recombinant eosinophil cationic protein predicts allergic lung inflammation in a mouse model for asthma

Abstract

Heparan sulfate proteoglycans (HSPGs) are glycoproteins consisting of a core protein to which linear heparan sulfate (HS) side chains are covalently attached. These HS side chains mediate a variety of biologic functions involved in inflammation. Radionuclide imaging of HS side chains in tissues with inflammation may be used for the stratification of patients who would most likely benefit from HSPG-targeting therapy. The goal of this study was to evaluate the feasibility of in vivo radionuclide imaging of HS side chain expression in a mouse model of asthma using the recombinant eosinophil cationic protein (rECP). Methods: rECP was radioiodinated with 125I or 123I using the Chloramine-T method. The 50% inhibitory concentration value for 125I-labeled rECP was determined in a competitive cell-binding assay using Beas-2B cells. The binding of radiolabeled rECP to HS side chains was evaluated both in vitro and in vivo. The biodistribution of radiolabeled rECP was assessed in asthma mice or in control mice using SPECT imaging, ex vivo biodistribution measurements, and microautoradiography. Results: The 50% inhibitory concentration value for 125I-rECP was 7.4 6 ± 0.1 nM. The loss of HS side chains substantially inhibited the cellular and tissue uptake of 125I- or 123I-rECP, indicating that HS side chains of HSPGs are required for 125I- or 123I-eosinophil cationic protein binding and uptake both in vitro and in vivo. SPECT imaging demonstrated an appreciably higher accumulation of radioactivity in the lungs of asthma mice than in those of control mice. Ex vivo biodistribution studies also confirmed that there was at least a 4-fold increase in the lung-to-muscle ratio of asthma mice, compared with control mice. The accumulation of radiolabeled rECP was linearly correlated with leukocyte infiltration. Conclusion: This study illustrates the feasibility of using radiolabeled rECP for the visualization of HS side chains of HSPGs and the evaluation of allergic lung inflammation in living subjects. Our data indicate that radiolabeled rECP is a novel imaging agent for HS side chains of HSPGs in predicting allergic lung inflammation in living mice. Copyright © 2013 by the Society of Nuclear Medicine and Molecular Imaging, Inc.