2. Victoria L. M. Herrera, M.D.
Associate Professor of Medicine
Section of Molecular Genetics
Whitaker Cardiovascular Institute
Boston University School of Medicine
700 Albany Street – W609, Boston MA 02118
vherrera@bu.edu
3. Tg53 rat model – “is it promising enough?”
Question analyzed –
• a provocative but necessary question posed by
Symposium organizers
• modeling the “vulnerable plaque” has been an
underestimated challenge
• a review of multi-species animal models of
atherosclerosis from the 1960s underscores this
challenge
4. Modeling vulnerable plaque – an underestimated challenge
Analysis of coronary artery
disease at end-stage
plq
hge
plq
eros’n
plq
fissure
plq
thombosis occlus’n
Pigeon: Carneau, Show Racer
* aortic root
nr + nr nr +
Monkey: a. Cynomolgus
b. African green
c. Rhesus
+
+
nr
nr
nr
nr
nr
nr
nr
nr
nr
nr
+
+
Pig: a. thiouracil + xrad’n
b. FHC: hyperLDL-emia
+
+
nr
nr
+
nr
some
some
+
+
Rabbit: WatanabeHHL
* coronary ostia
nr nr nr nr +
Mouse: ApoE/LDLr knockout nr nr nr nr +
Rat: Tg53 rat model + + + + +
[nr, not reported; +, present]
5. Elements of a “promising model”
Simulation of coronary artery disease
– plaque site
– disease course
– plaque progression
Model advantages as experimental system
– reproducibility
– duration to valid endpoints
– defined genetic background
– accessible phenotype manipulation
Model value and potential
– as investigative instrument
– as pre-clinical platform
6. Elements of a “promising model”
Tg53 rat model simulates human coronary heart
disease (CHD)
plaque site
plaque predilection site = coronary arteries
in contrast to: aortic root plaque predilection site in
mouse models described to date
Tg53 end-stage
plaques in proximal
right coronary artery
[PTAH stain, 40x
original mag]
RV
Ao
7. Elements of a “promising model”
Tg53 rat model simulates human coronary heart
disease (CHD)
disease course
risk factors consistent with human CHD
• hypertension exacerbates phenotype
• atherogenic lipid profile: increased total
cholesterol and triglyceride levels, low HDL,
increased VLDLc and VLDLtg, increased small
LDLc
• hyperlipidemia increases with age
8. Elements of a “promising model”
Tg53 rat model simulates human coronary heart
disease (CHD)
disease course
decreased survival compared with non-transgenic –
non-hyperlipidemic, hypertensive, Dahl S rat
controls on regular rat chow [Nat Med 5: 383, 1999; Mol
Med 7:831, 2001].
“end-stage” simulates cardiac endpoints of CHD
• (+) cardio-respiratory distress
• (+) signs of heart failure
9. Elements of a “promising model”
Tg53 rat model simulates human coronary heart
disease (CHD)
plaque progression
“culprit” plaque features: foam-cell rich, paucity of smcs, leukocyte
adhesion, intraplaque thrombi, erosion, plaque shoulder
susceptibility
Masson – trichrome stain 400x orig mag PTAH; 1000x orig
mag
10. Elements of a “promising model”
Masson – trichrome; 1000x orig
mag
Tg53 rat model simulates human coronary heart
disease (CHD)
plaque progression
“culprit plaque features: foam-cell rich, intraplaque hemorrhage,
paucity of smcs, lipid core > 1/3 of plaque volume
smc α-actin, Fast red im-stn; 400x
11. Elements of a “promising model”
Tg53 rat model simulates human coronary heart
disease (CHD)
plaque progression
“culprit” plaque features: fibrin-positive thrombosis in
proximal coronary lesions; none detected in more distal
stable lesions
PTAH: fibrin(+) thrombus; prox lesion PTAH: distal stable lesion
12. Elements of a “promising model”
Tg53 rat model simulates human coronary heart
disease (CHD)
plaque progression
recapitulates lesion heterogeneity of “culprit” plaques
associated with human acute coronary syndromes
PTAH: thick cap, smc-rich, but with
intraplq hge & thrombosis, IEL disrup’n
PTAH: reduced cap & smc; foam-cell
rich; + intraplq hge & thrombosis,
13. Elements of a “promising model”
Tg53 rat model: advantages as experimental system
reproducibility and robustness of phenotype
• lipid profile: increased total cholesterol and
triglycerides predominantly in VLDL, low HDL
• “culprit” plaque phenotype in proximal right coronary
artery
• “stable” occlusive plaque in smaller coronary arteries
reasonable duration to valid endpoints
• 6-9+ month range of proximal coronary plaque
development on regular rat chow: eccentric
macrophage-foam cell rich plaque which progresses to
“culprit” plaque at endstage
14. Elements of a “promising model”
Tg53 rat model: advantages as experimental system
defined genetic background – eliminates
differential genetic susceptibility as confounder
• inbred Dahl salt-sensitive hypertensive rat strain
accessible phenotype manipulation – onset and
course of hyperlipidemia and coronary artery
disease can be experimentally manipulated
• Western Type Diet (0.15% cholesterol) accelerates
onset and levels of hyperlipidemia
• low salt diet (0.0038% NaCl), which reduces level of
hypertension, attenuates coronary artery disease hence
increasing survival
15. Elements of a “promising model”
Tg53 rat model: value and potential
identical genetic background and ability to
regulate environmental factors allows well-
controlled in vivo studies for cross-talk
pathogenesis:
a. investigation of mechanisms of coronary
plaque development not possible in humans
b. methodical in vivo investigation of novel
intervention targets
c. maximization of genomic-based technologies
aimed at the investigation of mechanisms and
targets
16. Elements of a “promising model”
Tg53 rat model: value and potential
coronary-specific experimental design for
coronary artery-disease studies: with cumulative
evidence of vessel-specific genetic factors, the
Tg53 coronary-specific phenotype is key
combinatorial modeling: well characterized
inbred rat strains allow cross-breeding for
interaction studies relevant to human disease
hypertension
• diabetes
• obesity
17. Elements of a “promising model”
Tg53 rat model: value and potential
as investigative tool: invaluable new insight
distinct from that observed in current other models
– as a beginning,
1. prelude to culprit plaque: the eccentric macrophage
foam-cell rich lesion becomes the “culprit” plaque, thus
experimentally “capturing” the beginning of the
vulnerable plaque
2. differential pathway hypothesis: proximal vulnerable-
culprit coronary lesions develop distinct from more
distal stable coronary plaques
3. lesion heterogeneity paradigm: given identical genetic
background and environmental factors, lesion
heterogeneity at end-stage reflect stochastic endpoints
of a common pathogenic framework
18. Elements of a “promising model”
Tg53 rat model: value and potential
as pre-clinical platform
• identical genetic background
• regulated environmental factors
• robust coronary phenotype
• instrumentation accessibility
• rat physiological and pharmacological information
database
– allow continuity of comparative analysis for
successful new intervention and prevention
strategies
19. • ... can’t promise the world – but along with
other animal models, can together make a
better promise,
• but then again, which model can be
“promising enough” since even humans –
although they are the ultimate benchmarks –
make terrible models, if not the worst, for
human coronary artery disease ...
Tg53 rat model – “is it promising enough?”
20. Acknowledgment
• This work is supported by grants from the
National Institutes of Health and American
Heart Association.