SHAPE Society

1. Gene Array Study of Vulnerable
Atherosclerotic Plaque
In Apo E K/O Mice Plaques Prompted to
Rupture by Modified Falk Procedure
In Human Carotid Endarterectomized
Plaques
In Human Ruptured Coronary Arteries

2. Morteza Naghavi, MD
Sadaf Anwar, MD
Mohammad Madjid, MD
Silvio Litovsky, MD
Rahul Mitra, MD
Hugh A. McAllister,MD
Frank D. Kolodgie, MD
Ward Casscells, MD
James T Willerson, MD
University of Texas-Houston
Texas Heart Institute

25. Rationales of the Study
• The basic mechanisms leading to plaque rupture in
terms of genomic changes are still in shadow
• There has been no animal of plaque rupture to date
• An animal model offers a great opportunity to test
new diagnostic and therapeutic approaches
• Plaque vulnerability definition is at the center of
doubt and argument. Gene studies may help in
clearing this issue too.

26. Why Going to Gene Level?
Gene
Protein
Function
Sensitivity

27. Why is novel about this study?
•We are going to gene level, hence
having a higher probability of
detecting changes of interest
•We are using our rare clone of old
(>2yr) Apo E K/O mice

28. Of Mice and Human
• We are looking into three models of:
Apo E K/O Mice
Human carotid plaques
Human coronary plaques

29. Examples of Genes of Interest:
Apoptosis Lipid
Metabolism
Interleukins MMPs
Thrombosis Aspartate
Endopeptidases
NO system MMPIs
Cell
Adhesion
Molecules
Cysteine
Endopeptidases
Nuclear
factors and
receptors
Cathepsins
… Serine
Endopeptidases
… ….

30. 50 Apo E K/O mice age > 2 y
• Group A
45 mice for prompting to plaque rupture
• Group B
5 mice for control

31. Each Five Mice:
• L-NAME (N-nitro-L-arginine methyl ester)
(vasoconstriction)
• Cocaine (Hypertension, tachycardia)
• Xanthine + xanthine oxidase (inflammatory response)
• IL-1 beta (inflammatory response /septic shock)
• Adrenaline (hypertension, tachycardia)
• Methionine (direct cytotoxic effects destabilizing the
plaque)
• Buthionine sulfoximine (GSH inhibition, hypertension)

32. Multi-Factor Approach
Cocaine
+ xanthine
+ xanthine oxidase
+ adrenaline

33. 5 Mice are kept as controls
with no injection

34. At Day Seven:
• Mice are euthenized (Co2) and then
dissected
• Before day 7 Animals will be
monitored every 6 hours. If any of
them is found dead in the middle
of the experiment it will be
assessed for evidence of plaque
rupture

35. • Choice: specialized device for cross-
sectional cutting of total mouse body
• If not available: mice are dissected and
atherosclerotic plaque from aortic root
and arch from all mice are taken for
histopathological assessment as well as
mRNA study.