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Top Mouse
Models for
Cardiovascular
Disease Research
Technical Information Services
Upcoming JAX Webinars™
2
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The Jackson Laboratory’s Mission
4
Performing Research
Investigating genetics and biology of human disease
Providing Resources
JAX® Mice Clinical & Research Services, bioinformatics data,
technical publications and more…
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JAX® Mice The Gold Standard for Biomedical Research
JAX® Mice The Gold Standard for Biomedical Research
5
NIH funded resource
>8,000 strains and growing
o 2.7 million mice shipped annually
Unsurpassed genetic quality & animal health
Best characterized & referenced ~100 new pubs/week
Common inbred strains (C57BL/6J, BALB/cJ, DBA/2J) support
development/collection of specialty strains and other valuable
community research resources
JAX® Mice |
Online Resources to Expedite
Research
6
JAX® Mice Database www.jax.org/jax-mice-and-services/
Mouse Genome Informatics www.informatics.jax.org
Mouse Phenome Database phenome.jax.org/
And many more unique
resources
Contact Technical Support |
Online Resources to Expedite
Research
Cardiovascular Disease
7
Leading cause of morbidity
and mortality worldwide
(WHO)
Costs in the US
o $315 billion in 2010
o Projected to cost $2.8 trillion
by year 2030 (AHA 2014
update)
Coronary heart disease (Ischemic heart disease)
Cerebrovascular disease (stroke)
Peripheral arterial disease
Hypertension
Hypertrophy and Heart failure
Rheumatic heart disease
Congenital heart disease American
Heart
Association
Mouse Models for Cardiovascular
Disease
Coronary heart disease
o Atherosclerosis
o Ischemic/reperfusion injury & myocardial
infarction
Hypertension
Hypertrophy and heart failure
Other heart abnormalities
JAX® Mice | 8
JAX® Mice | 9
Sex Difference in Cardiovascular Disease Models
Sex differences in cardiovascular morbidity and mortality have long been
recognized. The mechanism is believed due largely to protective actions of
female hormones
Female mice are more tolerant to many cardiovascular injuries or stresses
than males
Do not mixed males and females in the same group
Mahmoodzadeh et al. 2012. Sex and Gender Differences in Pharmacology (214):23-48. [PMID: 23027444]
Mouse Models for
Cardiovascular Disease
Coronary heart disease
o Atherosclerosis
o Ischemic/reperfusion injury & myocardial
infarction
Hypertension
Hypertrophy and heart failure
Other heart abnormalities
JAX® Mice | 10
Atherosclerosis: Fatty artery wall deposits harden and block blood flow
Progressive disease
~ half a million people die
each year from the disease
Can lead to angina, heart
attack or stroke
Risk factors:
o High cholesterol, smoking, high
blood pressure, diabetes,
obesity and stress
http://www.nhlbi.nih.gov/health/dci/Diseases/Cad/CAD_WhatIs.html
JAX® Mice | 11
Development of Atherosclerotic
Plaques
JAX® Mice |
Stage 1 Stage 2 Stage 3 Stage 4
Rader et al. 2008. Nature 451(7181):904-13. [PMID: 18288179]
12
Lipoprotein Metabolism and
Transport
JAX® Mice |
Rader et al. 2008. Nature 451(7181):904-13. [PMID: 18288179]
13
The Mouse as a Model for Human
Atherosclerosis
Advantages
Lipoprotein metabolism and transport pathways similar to humans
C57BL/6J susceptible to atherosclerotic lesions
Develop lesions on Atherogenic or Western diet
o BUT… lesions do not progress past fatty streak stage
o Can be genetically manipulated to develop atherosclerosis
Considerations
Lesions require several months to develop
Most plasma cholesterol stored in HDL, not LDL
JAX® Mice | 14
Mouse Nomenclature
B6.129P2-Apoetm1Unc/J (002052)
Hypercholesterolemic (~ 400 mg/dl)
Increased circulating VLDL and LDL cholesterol
Decreased circulating HDL cholesterol
Spontaneously develop arterial lesions
o 3 months: fatty streaks in proximal aorta
o 13 months: atherosclerotic lesions
Ishida T et al. 2004. J Biol Chem 279(43):45085-92 [PMID: 15304490]
Moghadasian M et al. 2001. FASEB J 15(14):2623-30 [PMID: 11726538]
Ma Z et al. 2008. Clin Immunol 127(2):168-75 [PMID: 18325838]
JAX® Mice |
Spontaneous Atherosclerosis in ApoE
Null Mice (Normal “Chow” Diet)
15
Other JAX atherosclerosis susceptible mice
JAX® Mice
B6.129P2-Apoetm1Unc/J (002052)
Aortic plaques at 8 months of age
Elevated total & HDL cholesterol
Elevated Triglycerides
ApoE -/- ApoE +/-
Genotype Total
Chol
HDL
Chol
Triglyc
Wild-type 88 78 73
ApoE -/- 750 270 282
16
Spontaneous Atherosclerosis in ApoE
Null Mice (Normal “Chow” Diet)
Diets-Induced Atherosclerosis
Atherogenic (Paigen) Diet*
15% butter fat
1-1.25% cholesterol
0.5% cholic acid
50% sucrose
20% casein
JAX® Mice |
Western Diet
21% butter fat
0.2% cholesterol
0% cholic acid
34% sucrose
19.5% casein
*Cholesterol 10-20 times normal, cholic acid is pro-inflammatory
17
JAX mice susceptible to diets-induced atherosclerosis
JAX mice relatively resistant to diets-induced atherosclerosis
Diet Induced Athersclerosis in B6J Atherosclerotic (Paigen) Diet
Cholesterol 200-300 mg/dl (B6 females on chow = 80-90 mg/ml)
Lesions form by 7 weeks, thickened arterial wall and cell proliferation
By 14 weeks, fatty streaks with lipid filled cells and cuboidal nuclei
Lesions form in ascending aorta/aortic sinus
Mice survive to 28-40 weeks on diet
Paigen et al. 1990. Arteriosclerosis 10(2):316-23. [PMID: 2317166]
Normal 7 weeks 14 weeks
18
Diet Induced Athersclerosis in B6J Monocytes Exacerbates & IL-10 Alleviates Lesion
Development
Walker et al. 1999. Arterioscler Thromb Biol. 19(11):2673-9. [PMID: 10559010]
Mallat et al. 1999. Circ Res. 85(8):17-24. [PMID: 10521249]
16 weeks (Sirius Red/Collagen) 9 weeks (CD11b+ monocytes/red)
treated untreated IL-10 -/- IL-10 +/+
3-Deazaadenosine Treatment
Inhibition of monocyte chemotaxis & adhesion
Decreased number and size of lesions in
treated mice (fewer monocytes in lesions)
Normal B6 vs B6 IL-10 -/- (002251)
IL-10 is protective for lesion development
Increased number and size of lesions in the
IL-10 -/- mice
19
Atherogenic Diet (14 weeks)
Normal Diet (14 weeks)
Plaques on the aortic luminal
surface of 24 week old mice
B6.129P2-Apoetm1Unc/J (002052)
(15.8% fat, 1.25% cholesterol, 0.5% Na cholate)
Abundant, large plaques by 14 weeks
Increased plasma VLDL cholesterol
Increased plasma total cholesterol
Guo Y et al. 2005. Hear Res 208(1-2):54-67 [PMID: 16051453]
Ishida T et al. 2004. J Biol Chem 279(43):45085-92 [PMID: 15304490]
Ma Z et al. 2008. Clin Immunol 127(2):168-75 [PMID: 18325838]
JAX® Mice |
Atherosclerosis in ApoE Null Mice Accelerated Disease on an Atherogenic “Paigen” Diet
20
Apoe +/+
Apoe -/-
(athero diet)
Apoe -/-
Rati
o o
f in
tim
a t
o m
ed
ia (
I/M
)
Apoe +/+ Apoe -/- Apoe -/-
(athero diet) aortic sinus ascending aorta
Guo Y et al. 2005. Hear Res 208(1-2):54-67 [PMID: 16051453]
http://www.web-books.com/eLibrary/Medicine/Physiology/Cardiovascular/artery.jpg
JAX® Mice |
Atherosclerosis in ApoE Null Mice Accelerated Disease on an Atherogenic “Paigen” Diet
21
Ishibashi et al. 1994. J Clin Invest. 93(5):1885-93. [PMID: 8182121]
JAX® Mice |
Atherosclerosis in Ldlr Null Mice Atherogenic “Paigen” Diet Required for Atherosclerosis
Comparison of Normal Chow to Atherogenic Diet
B6.129S7-Ldlrtm1Her/J (002207)
Elevated total cholesterol on normal chow, no atherosclerosis
Hypercholesterolemia on atherogenic diet, increased VLDL and decreased HDL
22
Cholesterol
Ishibashi et al. 1994. J Clin Invest. 93(5):1885-93. [PMID: 8182121]
JAX® Mice |
Atherosclerosis in Ldlr Null Mice Atherogenic “Paigen” Diet Required for Atherosclerosis
B6.129S7-Ldlrtm1Her/J (002207)
No gross lesions in aorta of wild-type or
Ldlr -/- fed normal chow
Only Ldlr -/- fed atherogenic high
cholesterol diet showed plaques
Ldlr -/- fed atherogenic diet had severe
xanthomas
13 months of age, 8 months on diet
23
New Atherosclerosis Model “E4H”
Mouse Apoe replaced with human apoE4 isoform
Mouse Ldlr replaced with human LDLR
Highly sensitive to diet-induced dyslipidemia
Hypercholesterolemia
Atherosclerosis
Malloy SI et al. 2004. Arterioscler Thromb Vasc Biol 24 (1) 91-7 [PubMed: 12969990]
24
B6.129-Apoetm3(APOE*4)Mae Ldlrtm1(LDLR)Mae/J (012307)
On a high fat diet:
Mouse Models for
Cardiovascular Disease
Coronary heart disease
o Atherosclerosis
o Ischemic/reperfusion injury & myocardial
infarction
Hypertension
Hypertrophy and heart failure
Other heart abnormalities
JAX® Mice | 25
JAX® Mice | 26
Ischemia/Reperfusion (I/R) Injury and
Myocardial Infarction (MI)
Image created by J. Heuser, 2006
Acute MI is also known as a “heart attack”
Infarction: tissue death
Ischemia: lack of blood and oxygen supply
Reperfusion: restore of blood supply
Infarction area of the tip of the
anterior wall of the heart
Atherosclerosis
Thrombosis
Coronary
artery spasm
Coronary
occlusion
JAX® Mice | 27
In Vivo Experimental Models for I/R
or MI
Complied from Terese Winslow, Lydia Kibiuk 2001
Ligation of the left anterior descending (LAD)
coronary artery
Coronary artery occlusion
Readouts
Infarct size
Functional (or contractility)
recovery
Biomarkers for tissue damage Troponin I/T, CK and LDH activity, etc.
JAX® Mice | 28
I/R or MI Models Genetic Background Difference
FVB/NJ and ICR are resistant to I/R injury (increase occlusion time)
C57BL/6J and B6/129Sv/D2 are susceptible to I/R injury (decrease occlusion time)
Guo et al. 2012 Basic Res Cardiol. [PMID: 22864681]
Mouse Models for
Cardiovascular Disease
Coronary heart disease
o Atherosclerosis
o Ischemic/reperfusion & myocardial
infarction
Hypertension
Hypertrophy and heart failure
Other heart abnormalities
JAX® Mice | 29
Hypertension
Definition:
o systolic BP of 140 mmHg or greater
o diastolic BP of 90 mmHg or greater
> 50 million Americans suffer from hypertension
Increases risk of coronary heart disease, congestive heart failure, stroke and kidney disease
Contributing factors:
o Obesity
o Salt sensitivity
o Insulin resistance
o Genetic factors
o Age
JAX® Mice | 30
Inbred Strains for BP Research
Mouse Nomenclature
BPH/2J (003005) Hypertensive
BPL/1J (003006) Hypotensive
BPN/3J (003004) Normotensive
BPH vs. BPL & BPN
Elevated systolic BP by 5 weeks
~ 60 mmHg higher by 5 months
Elevated heart rates, enlarged hearts and kidneys, and higher
hematocrit
3-5 genes contributing to BP differences
Data in Mouse Phenome Database (MPD)
Schlager G and Sides J. 1997. Lab Animal Sci 47(3):288-92. [PMID: 9241632]
Uddin M et al. 2003. Mech Ageing Dev 124(7):811-7 [PMID: 12875744]
JAX® Mice | 31
B6 Agtr1a Null
Mouse Nomenclature
B6.129P2-Agtr1atm1Unc/J (002682)
Angiotensin 2 receptor 1a
Regulates sodium and fluid
homeostasis
Arterial pressure reduced by
~43 mmHg
Systolic BP reduced by ~24
mmHg
BP sensitive to [Na+]
Ito M et al. 1995. PNAS 92(8):3521-5 [PMID: 7724593]
JAX® Mice | 32
Mouse Models for
Cardiovascular Disease
Coronary heart disease
o Atherosclerosis
o Ischemic/reperfusion & myocardial
infarction
Hypertension
Hypertrophy and heart failure
Other heart abnormalities
JAX® Mice | 33
Hypertrophy and Heart Failure
JAX® Mice | 34
Hypertrophic cardiomyopathy
o The second most common form of heart muscle disease
o Genetically transmitted or induced
Heart failure
o Heart muscle weakened and cannot pump enough blood to the rest of the body
o Caused by conditions that damaged the heart muscle or overwork the heart
00
Decompensation
00 Figure compiled from Abcam.com
Atherosclerosis
Hypertension
Myocardial infarction
Other injuries or stress
Heart overworked
weakened and
enlarged
Hypertrophy and Heart Failure Common Experimental Models
JAX® Mice | 35
Jianyong et al. 2014 Evid Based Complement Alternat Med. [PMID: 25093027]
deAlmeida et al. 2010 J Vis Exp (38): 1729. [PMID: 3164086]
Weinheimer et al. 2015. Clin Exp Pharmacol Physiol 42(1):33-40 [PMID: 25311547]
Crowley et al. 2006 PNAS 103(47):17985-90. [PMID: 17090678]
Transverse Aortic Constriction (TAC):
Permanent MI (or combination of TAC and MI)
Angiotensin II infusion Readouts
Heart weight
Heart function
Heart wall thickness
Chamber size
Fibrosis
TAC Sham TAC position
B6 substrains Different Responses to TAC-induced Hypertrophy
JAX® Mice | 36
B6 substrains compared
o C57BL/6J (JL)
o C57BL/6NCrl (CL)
o C57BL/6NTac (TF)
Garcia-Menendez et al., 2013 Am J Physiol Heart Circ Physiol. 305(3): H397–H402. [PMID: 23709599]
JAX® Mice | 37
C57BL/6J is more tolerant to TAC-
induced hypertrophy
o Better survival
o Better heart function
o Less increase in heart weight
o Less wall thickness and dilation
Garcia-Menendez et al., 2013 Am J Physiol Heart Circ Physiol. 305(3): H397–H402. [PMID: 23709599]
B6 substrains Different Responses to TAC-induced Hypertrophy
C57BL/6J may more useful in studies
o Longer time
o Factors that make accelerate and
acerbate hypertrophy
B6 Nos3 Null (eNOS)
Mouse Nomenclature
B6.129P2-Nos3tm1Unc/J (002684)
C57BL/6J background
~140 mm Hg (~20 mmHg higher BP than
controls) by 12–16 weeks or age
After TAC*, mutant hearts show only
modest and concentric cardiac hypertrophy
by 3 weeks
Impaired pulmonary angiogenesis
Increased left ventricle systolic pressure
Shesely EG et al. 1996. PNAS 93(23):13176-81 [PMID: 8917564]
Takimoto E et al. 2005. J Clin Invest 115(5):1221-31 [PMID: 15841206]
Han RN et al. 2004. Circ Res 94(8):1115-23 [PMID: 15016731]
JAX® Mice |
*Transverse Aortic Constriction
38
JAX® Mice | 39
D’Angelo et al., 1997 PNAS. 94(15):8121-6. [PMID: 9223325]
Cardiac-specific Gαq Transgenic
mice
Mouse Nomenclature
FVB/N-Tg(Myh6-Gnaq)40Gwd/J
(012460)
Gαq is regulated by an α-myosin
heavy chain (Myh6) promoter
Enlarged heart
Increased cardiomyocytes size
Severely compromised systolic
function
Overt heart failure Transgenic Non-transgenic sibling
Mouse Models for
Cardiovascular Disease
Coronary heart disease
o Atherosclerosis
o Ischemic/reperfusion & myocardial
infarction
Hypertension
Hypertrophy and heart failure
Other heart abnormalities
JAX® Mice | 40
JAX® Mice | 41
Other Cardiomyopathies
Arrhythmia
o Bradycardia
o Tachycardia
Aortic aneurysm
o B6.129-Fbn1tm1Hcd/J (012885)
o B6.129S4-Timp1tm1Pds/J (006243)
Metabolic Syndrome
o B6.Cg-Lepob Ldlrtm1Her/J (006906)
o BKS.Cg-Leprdb Nos3tm1Unc/RhrsJ (008340)
Vascular Defects
JAX® Mice | 42
Cardiovascular Tool Strains
Cre Recombinase Expressing:
o Smooth muscle
B6.129S6-Taglntm2(cre)Yec/J (006878)
o Inducible in cardiomyocytes
B6.FVB(129)-Tg(Myh6-cre/Esr1*)1Jmk/J (005657)
o Endothelium of developing and quiescent vessels
B6.FVB-Tg(Cdh5-cre)7Mlia/J (006137)
o Cardiac muscle cells
B6.FVB-Tg(Myh6-cre)2182Mds/J (011038)
Fluorescent Protein Expressing:
o mCherry expression in cardiomyocytes
B6;D2-Tg(Myh6*-mCherry)2Mik/J (021577)
o GFP in endothelial cells
STOCK Tg(TIE2GFP)287Sato/J (003658)
www.jax.org/phenome
JAX® Mice | 43
JAX® Mice | 44
Atherosclerosis
JAX® Mice | 45
Blood Pressure
JAX® Mice | 46
47
Common inbred & specialty
JAX® Mice strains
Basic, custom & complex
breeding capabilities and
speed congenics
Genome scanning
Genome Scanning
Cryopreservation & recovery
Compound efficacy testing
Thank you!
48
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