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RGD-Conjugated Human Ferritin Nanoparticles for Imaging Vascular Inflammation and Angiogenesis in Experimental Carotid and Aortic Disease

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Abstract

Purpose

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.

Procedures

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.

Results

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.

Conclusions

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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References

  1. Libby P (2002) Inflammation in atherosclerosis. Nature 420:868–874

    Article  PubMed  CAS  Google Scholar 

  2. Galis ZS, Sukhova GK, Lark MW, Libby P (1994) Increased expression of matrix metalloproteinases and matrix degrading activity in vulnerable regions of human atherosclerotic plaques. J Clin Invest 94:2493–2503

    Article  PubMed  CAS  Google Scholar 

  3. Sukhova GK, Shi GP, Simon DI, Chapman HA, Libby P (1998) Expression of the elastolytic cathepsins S and K in human atheroma and regulation of their production in smooth muscle cells. J Clin Invest 102:576–583

    Article  PubMed  CAS  Google Scholar 

  4. Shimizu K, Mitchell RN, Libby P (2006) Inflammation and cellular immune response in abdominal aortic aneurysms. Arterioscler Thromb Vasc Biol 26:987–994

    Article  PubMed  CAS  Google Scholar 

  5. Lindholt JS, Vammen S, Fasting H, Henneberg EW, Heickendorff L (2000) The plasma level of matrix metalloproteinase 9 may predict the natural history of small abdominal aortic aneurysms. A preliminary study. Eur J Vasc Endovasc Surg 20:281–285

    Article  PubMed  CAS  Google Scholar 

  6. Virmani R, Kolodgie FD, Burke AP et al (2005) Atherosclerotic plaque progression and vulnerability to rupture: angiogenesis as a source of intraplaque hemorrhage. Arterioscler Thromb Vasc Biol 25:2054–2061

    Article  PubMed  CAS  Google Scholar 

  7. Tedesco MM, Terashima M, Blankenberg FG et al (2009) Analysis of in situ and ex vivo vascular endothelial growth factor receptor expression during experimental aortic aneurysm progression. Arterioscler Thromb Vasc Biol 29:1452–1457

    Article  PubMed  CAS  Google Scholar 

  8. Saraff K, Babamusta F, Cassis LA, Daugherty A (2003) Aortic dissection precedes formation of aneurysms and atherosclerosis in angiotensin II-infused. Apolipoprotein E deficient mice. Arterioscler Thromb Vasc Biol 23:1621–1426

    Article  PubMed  CAS  Google Scholar 

  9. Paik DC, Fu C, Bhattacharya J, Tilson MD (2004) Ongoing angiogenesis in blood vessels of the abdominal aortic aneurysm. Exper Mol Med 36:524–533

    CAS  Google Scholar 

  10. Choke E, Thompso MM, Dawson J et al (2006) Abdominal aortic aneurysm rupture is associated with increased medial neovascularization and overexpression of proangiogenic cytokines. Arterioscler Thromb Vasc Biol 26:2077–2082

    Article  PubMed  CAS  Google Scholar 

  11. Hoshiga M, Alpers CE, Smith LL, Giachelli CM, Schwartz SM (1995) Alpha-v beta-3 integrin expression in normal and atherosclerotic artery. Circ Res 77:1129–1135

    PubMed  CAS  Google Scholar 

  12. Antonov AS, Kolodgie FD, Munn DH, Gerrity RG (2004) Regulation of macrophage foam cell formation by alphaVbeta3 integrin: potential role in human atherosclerosis. Am J Pathol 165:247–258

    Article  PubMed  CAS  Google Scholar 

  13. Waldeck J, Häger F, Höltke C et al (2008) Fluorescence reflectance imaging of macrophage-rich atherosclerotic plaques using an alphavbeta3 integrin-targeted fluorochrome. J Nucl Med 49:1845–1851

    Article  PubMed  Google Scholar 

  14. Laitinen I, Saraste A, Weidl E et al (2009) Evaluation of alphavbeta3 integrin-targeted positron emission tomography tracer 18F-galacto-RGD for imaging of vascular inflammation in atherosclerotic mice. Circ Cardiovasc Imaging 2:331–338

    Article  PubMed  Google Scholar 

  15. Winter PM, Morawski AM, Caruthers SD et al (2003) Molecular imaging of angiogenesis in early-stage atherosclerosis with alpha (v) beta3-integrin-targeted nanoparticles. Circulation 108:2270–2274

    Article  PubMed  CAS  Google Scholar 

  16. Liepold L, Anderson S, Willits D et al (2007) Viral capsids as MRI contrast agents. Magn Reson Med 58:871–879

    Article  PubMed  CAS  Google Scholar 

  17. Hooker JM, Datta A, Botta M, Raymond KN, Francis MB (2007) Magnetic resonance contrast agents from viral capsid shells: a comparison of exterior and interior cargo strategies. Nano Lett 7:2207–2210

    Article  PubMed  CAS  Google Scholar 

  18. Anderson EA, Isaacman S, Peabody DS, Wang EY, Canary JW, Kirshenbaum K (2006) Viral nanoparticles donning a paramagnetic coat: conjugation of MRI contrast agents to the MS2 capsid. Nano Lett 6:1160–1164

    Article  PubMed  CAS  Google Scholar 

  19. Uchida M, Terashima M, Cunningham CH et al (2008) A human ferritin iron oxide nano-composite magnetic resonance contrast agent. Magn Reson Med 60:1073–1081

    Article  PubMed  CAS  Google Scholar 

  20. Uchida M, Flenniken ML, Allen M et al (2006) Targeting of cancer cells with ferrimagnetic ferritin cage nanoparticles. J Am Chem Soc 128:16626–16633

    Article  PubMed  CAS  Google Scholar 

  21. Aime S, Frullano L, Geninatti, Crich S (2002) Compartmentalization of a gadolinium complex in the apoferritin cavity: a route to obtain high relaxivity contrast agents for magnetic resonance imaging. Angew Chem Int Ed 41:1017–1019

    Article  CAS  Google Scholar 

  22. Flenniken ML, Willits DA, Brumfield S, Young MJ, Douglas T (2003) The small heat shock protein cage from methanococcus jannaschii is a versatile nanoscale platform for genetic and chemical modification. Nano Lett 3:1573–1576

    Article  CAS  Google Scholar 

  23. Liepold LO, Abedin MJ, Buckhouse ED, Frank JA, Young MJ, Douglas T (2009) Supramolecular protein cage composite MR contrast agents with extremely efficient relaxivity properties. Nano Lett 9:4520–4526

    Article  PubMed  CAS  Google Scholar 

  24. Terashima M, Uchida M, Kosuge H et al (2011) Human ferritin cages for imaging vascular macrophages. Biomaterials 32:1430–1437

    Article  PubMed  CAS  Google Scholar 

  25. Uchida M, Willits DA, Muller K et al (2009) Intracellular distribution of macrophage targeting ferritin–iron oxide nanocomposite. Adv Mater 21:458–462

    Article  CAS  Google Scholar 

  26. Arap W, Pasqualini R, Ruoslahti E (1998) Cancer treatment by targeted drug delivery to tumor vasculature in a mouse model. Science 279:377–380

    Article  PubMed  CAS  Google Scholar 

  27. Kułdo JM, Ogawara KI, Asgeirsdóttir SA et al (2005) Molecular pathways of endothelial cell activation for (targeted) pharmacological intervention of chronic inflammatory diseases. Curr Vasc Pharmacol 3:11–39

    Article  PubMed  Google Scholar 

  28. Terashima M, Ehara S, Yang E et al (2010) In vivo bioluminescence imaging of inducible nitric oxide synthase gene expression in vascular inflammation. Mol Imaging Biol. doi:10.1007/s11307-010-0451-5

    Google Scholar 

  29. Kumar A, Lindner V (1997) Remodeling with neointima formation in the mouse carotid artery after cessation of blood flow. Arterioscler Thromb Vasc Biol 17:2238–2244

    Article  PubMed  CAS  Google Scholar 

  30. Daugherty A, Manning MW, Cassis LA (2000) Angiotensin II promotes atherosclerotic lesions and aneurysms in apolipoprotein E-deficient mice. J Clin Invest 105:1605–1612

    Article  PubMed  CAS  Google Scholar 

  31. Pang JH, Jiang MJ, Chen YL et al (1996) Increased ferritin gene expression in atherosclerotic lesions. J Clin Invest 97:2204–2212

    Article  PubMed  CAS  Google Scholar 

  32. Li W, Xu LH, Forssell C, Sullivan JL, Yuan XM (2008) Overexpression of transferrin receptor and ferritin related to clinical symptoms and destabilization of human carotid plaques. Exp Biol Med 233:818–826

    Article  CAS  Google Scholar 

  33. Brooks PC, Clark RA, Cheresh DA (1994) Requirement of vascular integrin alpha v beta 3 for angiogenesis. Science 264:569–571

    Article  PubMed  CAS  Google Scholar 

  34. Ntziachristos V, Ripoll J, Wang LV, Weissleder R (2005) Looking and listening to light: the evolution of whole-body photonic imaging. Nat Biotechnol 23:313–320

    Article  PubMed  CAS  Google Scholar 

  35. Nahrendorf M, Waterman P, Thurber G et al (2009) Hybrid in vivo FMT-CT imaging of protease activity in atherosclerosis with customized nanosensors. Arterioscler Thromb Vasc Biol 29:1444–1451

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

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).

Disclosures

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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Authors and Affiliations

  1. Division of Cardiovascular Medicine, Stanford University School of Medicine, 300 Pasteur Drive, Room H2157, Stanford, CA, 94305–5233, USA

    Toshiro Kitagawa, Hisanori Kosuge & Michael V. McConnell

  2. Department of Chemistry, Montana State University, 112 Chemistry and Biochemistry Building, Bozeman, MT, 59715, USA

    Masaki Uchida & Trevor Douglas

  3. Division of Vascular Surgery, Stanford University School of Medicine, 300 Pasteur Drive, Stanford, CA, 94305, USA

    Monica M. Dua, Yasunori Iida & Ronald L. Dalman

Authors
  1. Toshiro Kitagawa
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  2. Hisanori Kosuge
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  3. Masaki Uchida
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  4. Monica M. Dua
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  5. Yasunori Iida
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  6. Ronald L. Dalman
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  7. Trevor Douglas
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  8. Michael V. McConnell
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Corresponding author

Correspondence to Michael V. McConnell.

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Supplemental Figure 1

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)

Supplemental Figure 2

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)

Supplemental Figure 3

. 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)

Supplemental Figure 4

. 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

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