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J Am Coll Cardiol, 2004; 43:453-460, doi:10.1016/j.jacc.2003.07.048
© 2004 by the American College of Cardiology Foundation
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Intravascular ultrasound molecular imaging of atheroma components in vivo

Andrew J. Hamilton, MD*,*, Shao-Ling Huang, PhD{dagger}, Drew Warnick, BS*, Mark Rabbat, BS*, Bonnie Kane, BS*, Ashwin Nagaraj, PhD*, Melvin Klegerman, PhD{ddagger} and David D. McPherson, MD*

* Department of Medicine, Feinberg School of Medicine, Northwestern University, Chicago, Illinois, USA
{dagger} Department of Biochemistry, Molecular Biology and Cell Biology, Northwestern University, Evanston, Illinois, USA
{ddagger} EchoDynamics, College Park, Maryland, USA

* Reprint requests and correspondence: Dr. David D. McPherson, Department of Medicine, Division of Cardiology, Northwestern University, 251 East Huron, Galter 8-230, Chicago, Illinois 60611-2908, USA.
d-mcpherson{at}northwestern.edu

This study was presented in part at the American College of Cardiology Scientific Sessions, Atlanta, Georgia, March 17 to 20, 2002.

OBJECTIVES: Our purpose was to quantitate and confirm specific echogenic immunoliposome (ELIP) atheroma component enhancement in vivo.

BACKGROUND: Targeted ELIPs for ultrasonic detection and staging of active molecular components of endothelium and atheroma have been developed.

METHODS: In Yucatan miniswine, the endothelium was injured from one femoral and one carotid artery, and animals were fed a high-cholesterol diet for two months to create various stages of atheroma. Arteries were imaged with intravascular ultrasound (IVUS) 5 and 10 min after ELIP injection (5-mg dose). Anti-intercellular adhesion molecule-1 (ICAM-1), anti-vascular cell adhesion molecule-1 (VCAM-1), anti-fibrin, anti-fibrinogen, and anti-tissue factor (TF) conjugated ELIPs were used, and immunohistochemistry (IHC) confirmed the presence or absence of molecular expression. Two blinded observers determined if each segment was enhanced by ELIP. Three-dimensional image reconstruction and videodensitometric analysis determined the mean gray-scale (MGS) change of the luminal border.

RESULTS: To determine endothelial injury component enhancement, anti-fibrinogen ELIP enhanced exposed fibrin in all arteries (MGS increased 22 ± 5%; 6 arteries; 2 animals). To determine enhancement of molecular components in atherosclerotic arteries, observers detected enhancement 5 min after anti-VCAM, anti-ICAM, anti-TF, anti-fibrin, and anti-fibrinogen conjugated ELIPs. Furthermore, ELIP enhanced atheroma MGS by 39 ± 18% (n = 8). The IHC staining confirmed the expression of respective molecular targets in all enhanced segments.

CONCLUSIONS: It was shown that ELIPs specifically enhance endothelial injury/atheroma components. This allows better characterization of the type and extent of active atheroma components and may allow more directed therapy.

Abbreviations and Acronyms
  ELIP = echogenic immunoliposome
  ICAM-1 = intercellular adhesion molecule-1
  IHC = immunohistochemistry
  IVUS = intravascular ultrasound
  M(Ab) = (monoclonal) antibody
  MGS = mean gray scale
  PC = phosphatidylcholine
  PE = phosphatidylethanolamine
  PG = phosphatidylglycerol
  TF = tissue factor
  VCAM-1 = vascular cell adhesion molecule-1




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