Micro‐Imaging of Amyloid in Mice

Wall, Jonathan S.; Paulus, Michael J.; Gleason, Shaun S.; Gregor, Jens; Solomon, Alan; Kennel, Stephen J.

doi:10.1016/s0076-6879(06)12011-x

Cited by 29 publications

(24 citation statements)

References 56 publications

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“…Mice were positioned prone on a cardboard platform, and the board was placed on the scanner bed (46). CT data were acquired using an X-ray voltage biased to 80 kVp with a 500-μA anode current and the data binned at 4 × 4.…”

Section: Methodsmentioning

confidence: 99%

“…Samples of muscle, liver, spleen, pancreas, kidney, heart, tongue, stomach, small and large intestines, and lung tissue were harvested postmortem from every mouse (46). Each sample was placed into a tared plastic vial and weighed, and the 125 I radioactivity was measured using an automated Wizard 3 gamma counter (1480 Wallac Gamma Counter; PerkinElmer).…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

In vivo molecular imaging of peripheral amyloidosis using heparin-binding peptides

Wall

Richey²,

Stuckey

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

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106

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Heparan sulfate proteoglycans (HSPGs) are ubiquitous components of pathologic amyloid deposits in the organs of patients with disorders such as Alzheimer's disease or systemic light chain (AL) or reactive (AA) amyloidosis. Molecular imaging methods for early detection are limited and generally unavailable outside the United Kingdom. Therefore, there is an urgent need to develop novel, specific amyloidophilic radiotracers for imaging to assist in diagnosis, prognostication, and monitoring response to therapy. Amyloid-associated HSPG can be differentiated from HSPG found in surrounding healthy cells and tissues by the preferential binding of certain HS-reactive single chain variable fragments and therefore, represents a biomarker that can be targeted specifically with appropriate reagents. Using a murine model of AA amyloidosis, we have examined the in vivo amyloid reactivity of seven heparin-binding peptides by using single photon emission and X-ray computed tomographic imaging, microautoradiography, and tissue biodistribution measurements. All of the peptides bound amyloid deposits within 1 h post-injection, but the extent of the reactivity differed widely, which was evidenced by image quality and grain density in autoradiographs. One radiolabeled peptide bound specifically to murine AA amyloid in the liver, spleen, kidney, adrenal, heart, and pancreas with such avidity that it was observed in single photon emission tomography images as late as 24 h post-injection. In addition, a biotinylated form of this peptide was shown histochemically to bind human AA, ALκ, ALλ, transthyretin amyloidosis (ATTR), and Aβ amyloid deposits in tissue sections. These basic heparin-binding peptides recognize murine and human amyloid deposits in both in vivo and ex vivo tissues and therefore, have potential as radiotracers for the noninvasive molecular imaging of amyloid deposits in situ.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

In vivo molecular imaging of peripheral amyloidosis using heparin-binding peptides

Wall

Richey²,

Stuckey

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

106

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“…The mice were used 6 weeks after the injection of amyloid enhancing factor, at which time AA deposits are widespread in the almost all visceral organs and tissues. Antibody NS4F5 was labeled with iodine-125 ( 125 I) via production of iodotyrosine using chloramine T as the oxidizing agent (20). Free iodide was removed from the preparation chromatographically by using an Ultrogel AcA 34 column (Amersham Biosciences).…”

Section: Detection Of Apoptosis In Human Lung Epithelial Cells After mentioning

confidence: 99%

The Heparan Sulfate Motif (GlcNS6S-IdoA2S)3, Common in Heparin, Has a Strict Topography and Is Involved in Cell Behavior and Disease

Smits

Kurup

Rops³

et al. 2010

Journal of Biological Chemistry

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Heparan sulfate (HS) is a structurally complex polysaccharide that interacts with a broad spectrum of extracellular effector ligands and thereby is thought to regulate a diverse array of biologic processes. The specificity of HS-ligand interactions is determined by the arrangement of sulfate groups on HS, which creates distinct binding motifs. Biologically important HS motifs are expected to exhibit regulated expression, yet there is a profound lack of tools to identify such motifs; consequently, little is known of their structures and functions. We have identified a novel phage display-derived antibody (NS4F5) that recognizes a highly regulated HS motif (HS NS4F5 ), which we have rigorously identified as (GlcNS6S-IdoA2S) 3 . HS NS4F5 exhibits a restricted expression in healthy adult tissues. Blocking HS NS4F5 on cells in culture resulted in reduced proliferation and enhanced sensitivity to apoptosis. HS NS4F5 is up-regulated in tumor endothelial cells, consistent with a role in endothelial cell activation. Indeed, TNF-␣ stimulated endothelial expression of HS NS4F5 , which contributed to leukocyte adhesion. In a mouse model of severe systemic amyloid protein A amyloidosis, HS NS4F5 was expressed within amyloid deposits, which were successfully detected by microSPECT imaging using NS4F5 as a molecularly targeted probe. Combined, our results demonstrate that NS4F5 is a powerful tool for elucidating the biological function of HS NS4F5 and can be exploited as a probe to detect novel polysaccharide biomarkers of disease processes.Heparan sulfate proteoglycans (HSPGs), 3 major components of the cell surface and the extracellular matrix, are involved in a variety of biological phenomena, including cell adhesion, proliferation, differentiation, and inflammation as well as being associated with pathologic events such as atherosclerosis and amyloidosis. Because of their high negative charge, HS chains interact with a variety of proteins, including growth factors/ morphogens and their receptors, the amyloid precursor protein serum amyloid protein A (AA), chemokines, and extracellular matrix proteins. HS-protein interactions vary with regard to specificity and may depend on charge density in addition to strict sequence motifs of HS.The interaction of heparin and HS with FGFs and their receptors has been characterized in great detail. Specific HS structures are predominantly determined by the regulated positioning of N-, 2-, 6-, and 3-O-sulfate groups along HS chains (1). For example, FGF-2 requires both N-and 2-O-sulfate groups for binding to HS. The 6-O-sulfate group is not essential for binding to FGF-2 but is critical for activation of the FGF receptor (2). In contrast, binding of hepatocyte growth factor, platelet-derived growth factor, lipoprotein lipase, and herpes simplex virus glycoprotein C to HS is dependent on 6-O-sulfation (3). The activation of antithrombin III by HS/heparin is mediated by a specific pentasaccharide in which a 3-O-sulfate group is crucial (4). Thus, the biological functions of HSPGs are contro...

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“…Peptides (10–40 μg) were radioiodinated with 125 I (Perkin Elmer, Waltham, MA) using chloramine T (10 μg) and the products purified by gel filtration and shown to be>95% pure by SDS-PAGE followed by phosphorimager analysis as described elsewhere [36]. For SPECT/CT imaging studies, peptide p5 was radiolabeled with 99m Tc from pertechnitate (Cardinal Health, Knoxville, TN), using 100 ng of SnCl 2 according to the method of Tran et al [37].…”

Section: Methodsmentioning

confidence: 99%

The pattern recognition reagents RAGE VC1 and peptide p5 share common binding sites and exhibit specific reactivity with AA amyloid in mice

Kennel

Williams²,

Stuckey

et al. 2015

Amyloid

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In the US, there remains a need to develop a clinical method for imaging amyloid load in patients with systemic, visceral amyloidosis. The receptor for advanced glycation end products (RAGE), which exists as a transmembrane receptor and soluble variant, is found associated with a number of amyloid deposits in man. It is unclear whether amyloid-associated RAGE is the membrane or soluble form; however, given the affinity of RAGE for amyloid, we have examined the ability of soluble RAGE VC1 to specifically localize with systemic AA amyloid in mice. We further compared the reactivity of RAGE VC1 with that of the synthetic, amyloid-reactive peptide p5. Methods Binding of radiolabeled RAGE VC1 and p5 to synthetic amyloid fibrils was evaluated using in vitro “pulldown” assays in the presence or absence of RAGE ligands. Radioiodinated RAGE VC1 and technetium-99 m-labeled p5 were studied in mice with systemic AA amyloidosis using dual-energy SPECT/CT imaging, biodistribution and microautoradiography. Results Soluble RAGE VC1 competed with radioiodinated peptide p5 for binding to rVλ6Wil, Aβ (1–40) and IAPP fibrils but not with the higher affinity peptide, p5R. Pre-incubation with AGE-BSA abrogated binding of VC1 and p5 to rVλ6Wil fibrils. Dual-energy SPECT/CT images and quantitative tissue biodistribution data showed that soluble RAGE VC1 specifically bound AA amyloid-laden organs in mice as effectively as peptide p5. Furthermore, microautoradiography confirmed that RAGE VC1 bound specifically to areas of Congo red-positive amyloid in mouse tissues but not in comparable tissues from control WT mice. Conclusion Soluble RAGE VC1 and peptide p5 have similar ligand binding properties and specifically localize with visceral AA amyloid deposits in mice.

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Micro‐Imaging of Amyloid in Mice

Cited by 29 publications

References 56 publications

In vivo molecular imaging of peripheral amyloidosis using heparin-binding peptides

In vivo molecular imaging of peripheral amyloidosis using heparin-binding peptides

The Heparan Sulfate Motif (GlcNS6S-IdoA2S)3, Common in Heparin, Has a Strict Topography and Is Involved in Cell Behavior and Disease

The pattern recognition reagents RAGE VC1 and peptide p5 share common binding sites and exhibit specific reactivity with AA amyloid in mice

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