Inflammation and angiogenesis are important contributors to vascular disease. We evaluated imaging both of these biological processes, using Arg–Gly–Asp (RGD)-conjugated human ferritin nanoparticles (HFn), in experimental carotid and abdominal aortic aneurysm (AAA) disease.
Macrophage-rich carotid lesions were induced by ligation in hyperlipidemic and diabetic FVB mice (n = 16). AAAs were induced by angiotensin II infusion in apoE−/− mice (n=10). HFn, with or without RGD peptide, was labeled with Cy5.5 and injected intravenously for near-infrared fluorescence imaging.
RGD-HFn showed significantly higher signal than HFn in diseased carotids and AAAs relative to non-diseased regions, both in situ (carotid: 1.88 ± 0.30 vs. 1.17 ± 0.10, p = 0.04; AAA: 2.59 ± 0.24 vs. 1.82 ± 0.16, p = 0.03) and ex vivo. Histology showed RGD-HFn colocalized with macrophages in carotids and both macrophages and neoangiogenesis in AAA lesions.
RGD-HFn enhances vascular molecular imaging by targeting both vascular inflammation and angiogenesis, and allows more comprehensive detection of high-risk atherosclerotic and aneurysmal vascular diseases.
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We thank Dr. Matthew Bogyo and his laboratory for their assistance with mouse fluorescence molecular tomography. We thank support from the National Institutes of Health (R21 EB005364, R01 HL078678, P50 HL083800).
Dr. McConnell's laboratory receives research support from GE Healthcare and he is on a scientific advisory board for Kowa, Inc. Dr. Dalman's laboratory receives AAA research support from Medtronic AVE and Carolus Pharmaceuticals. The other authors have no potential conflicts of interest.
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Bio-distribution analysis of RGD-HFn and HFn based on ex vivo Cy5.5 signal. The distribution of nanoparticles in the major organs after 48 hours was similar between RGD+ (injected with RGD-HFn-Cy5.5) and RGD− (injected with HFn-Cy5.5) groups (p = NS RGD+ vs. RGD− for all organs) (JPEG 57 kb)
Macrophage immunofluorescence staining of representative non-ligated right carotid arteries in RGD− group. Immunofluorescence double staining for HFn-Cy5.5 and macrophages (stained with Mac-3) demonstrated very little macrophage infiltration and minimal Cy5.5 signal in non-diseased right carotid arteries (JPEG 42 kb)
. Smooth muscle cell (SMC) immunofluorescence staining of representative ligated left carotid arteries. RGD-HFn-Cy5.5 and HFn-Cy5.5 both showed limited colocalization to carotid SMCs (see text for quantitative colocalization analysis) (JPEG 105 kb)
. Smooth muscle cell (SMC) immunofluorescence staining of representative abdominal aortic aneurysms (AAA). RGD-HFn-Cy5.5 and HFn-Cy5.5 both showed limited colocalization to AAA SMCs (see text for quantitative colocalization analysis) (JPEG 88 kb)
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Kitagawa, T., Kosuge, H., Uchida, M. et al. RGD-Conjugated Human Ferritin Nanoparticles for Imaging Vascular Inflammation and Angiogenesis in Experimental Carotid and Aortic Disease. Mol Imaging Biol 14, 315–324 (2012). https://doi.org/10.1007/s11307-011-0495-1
- Vascular disease