During this presentation, Dr. Fred Beasley gives a broad-level overview of the factors to consider when deciding on an appropriate preclinical rodent model for studying obesity and its treatments. It features an overview of trends in obesity and associated illnesses, and the role of pharmacological intervention. Dr. Beasley discusses criteria for establishing a study’s aims and review commonly used rodent models obesity, addressing both genetically inherited and diet-induced obesity. The webinar concludes with additional considerations for improving your study design. Key topics include… - An introduction to the role of pharmacological intervention in treating obesity - A guide to establishing study aims for obesity research - An overview of commonly used rodent obesity models (diet and genetic) - Additional considerations for obesity research study design Key methods reviewed include… Pharmacotherapy, diet induced obesity, Western-style diet, monogenic obese rodents, polygenic obese rodents, NAFLD activity score

1. Fred Beasley, PhD
Director of Scientific Engagement,
Cardiovascular and Metabolic Disease
Crown Bioscience
Welcome to Obesity 2020
Animal Model Selection, Study Design
and Current Trends in Preclinical
Obesity Research

2. Dr. Fred Beasley provides an overview of study
design, criteria for defining your study aim and
expert advice for selecting the ideal rodent model
for your preclinical obesity research.
Welcome to Obesity 2020
Animal Model Selection, Study Design
and Current Trends in Preclinical
Obesity Research
Copyright 2020 Crown Bioscience, APS and InsideScientific. All Rights Reserved.

3. About the Speaker
3
• Dr. Fred Beasley, Director of Scientific Engagement
– PhD in Microbiology and Immunology, University of
Western Ontario
– Postdoctoral training in drug discovery and development
for infectious diseases and innate immunity, UC San
Diego and Calibr
• Joined CrownBio in 2019
• Extensive experience in animal model development
for pharmacology, inflammation, fibrosis,
bacteriology, and parasitology
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4. Overview
• Obesity and how pharmacological
intervention can help
• Establishing research aims
• Choose your route
– Dietary induction of disease
– Genetic basis for disease
• Additional considerations
• Summary
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5. • Obesity = “Body fat excess that causes
negative effects on health”
• Input vs. expenditure
– Food intake
– Physical activity
– Genetic factors affecting absorption,
metabolism, and storage
• Body mass index
(BMI)
5
body mass (kg)
height2 (m2)
Obesity Crisis: Growing Global Concern
• Adiposity will increase rates of
complications and comorbidities
– Metainflammation
– “Diabesity”
Category BMI
Overweight 25.0 – 29.9
Obese 30.0 – 34.9
Severely obese >35.0
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6. 6
Aune et al. BMJ 2016;353: i2156
Difference in cost/
100K obese
patients relative to
100K non-obese
patients (USA)
Obesity Crisis: Impact on Health
Mortality risk increases
with BMI >25
Blume et al. J Med Econ 2015;18: 1020-28
Comorbidities increase health care costs
associated with obese populations
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7. 7
• One pandemic on top
of another: obesity is
associated with
negative outcomes
after COVID-19
infection:
– Hospitalization, ICU
admission
– Need for ventilation
– ARDS and death
COVID patients requiring mechanical ventilation
Simmonet et al. Obesity 2020;28: 1195
Obesity Crisis: COVID Connection
Rottoli et al. Eur J Endocrinol 2020;183: 389
Risk of respiratory failure
Risk of ICU admission
Risk of death
Meta‐analysis of the association between individuals with
obesity and the risk of hospitalization with COVID‐19
Popkin et al. Obes Rev 2020;10.1111/obr.13128
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8. • Globally, obesity tripled since 1975
– 13% of global adult population
– Nearly 40% of US adult population
• Diet and physical activity should be
first-line treatment
• Persistent hormone changes
encourage weight regain
– Increased ghrelin; decreased peptide YY,
amylin, cholecystokinin
– Similar reports for leptin, GLP-1
8
overweight
obese
Men Women
NCD Risk Factor Collaboration. Lancet 2016;387: 1377
Sumithran et al. N Eng J Med 2011;365: 1597
Obesity Crisis: Future Concerns
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9. Current Pharmacological Landscape
9
NIH recommends pharmacotherapy for BMI over 30, or BMI of 27-30 with at least one comorbidity
FDA-approved anti-obesity drugs
Drug Term MOA Side Effects
Orlistat Chronic Lipase inhibitor, prevents absorption of dietary fat GI effects; steatorrhea; very rare cases of liver damage
Phentermine +
Topiramate
Chronic
• Stimulant, promotes epinephrine release
• Multiple targets, precise MoA unknown
Benign; paraesthesia, constipation, dry mouth
Bupropion +
Naltrexone
Chronic
• Norepinephrine and dopamine reuptake inhibitor; increases
piomelanocortin neuronal firing and suppresses appetite
• Opioid receptor antagonist; augments POMC activation
Nausea, constipation, vomiting
Liraglutide Chronic
GLP-1 analogue; increases insulin release and delays gastric emptying;
promotes satiety
Nausea, constipation, vomiting; possible cardiovascular
effects
Lorcaserin Chronic 5HT 2C (serotonin) receptor agonist; regulates appetite
Headache, nausea, fatigue
Increased cancer rate
Withdrawn Feb. 2020
Benzphetamine Short Sympathomimetic amine; stimulates central nervous system anorectic agent
Dry mouth, constipation, insomnia, dizziness, anxiety,
headache, raised blood pressure; potential for abuse
Diethylpropion Short Stimulant; promotes norepinephrine release; appetite suppressant
Phendimetrazine Short Stimulant; promotes norepinephrine release; appetite suppressant
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10. Establishing Research Aims
10
• Advantages of rodent models
– Conserved hormone/receptors systems, metabolic
enzymes relevant to human obesity
– Rapid disease development window
– Genetic tractability
– Convenient size, commercial commodity
• Acknowledge the limitations
– Restrictive enclosures
– Uncontrolled food intake
– Disregard for food choice
• Focus on the goal: What mechanism or process are
you targeting?
• Define your readouts: A measurable phenotype or
pathology with a good therapeutic window may suffice
Avert or resolve
obesity
Caloric intake vs
expenditure
Nutrient composition
Genetic determinants
Comorbidities
of obesity
Cardiovascular
disease
NAFLD/NASH
Beta cell failure
Nephropathy .
Links to
metabolic
disease
Hyperglycemia
Hyperlipidemia
Insulin resistance
Chronic
inflammation
www.jax.org
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11. 11
Preclinical rodent
model for drug testing
Diet-induced obesity
(DIO)
High fat diet
HFD + (sugar,
trans-/saturated fat,
cholesterol)
Genetic
Monogenic
Polygenic
Normal diet
High fat diet
HFD+
Model Selection
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12. Corporate Headquarters:
16550 West Bernardo Drive
Building 5, Suite 525
San Diego, CA 92127
Tel: 855.827.6968
Fax: 888.882.4881
www.crownbio.com
Diet-Induced Obesity Models

13. Diet-Induced Obesity: High Fat
13
• Go from low
energy density
to high energy
density diet
• Select obese (“DIO”) from lean non-responders (“DR”)
– Outbred rats (Wistar, Long Evans, Sprague Dawley): DIO rate 40-60%, M & F
– C57BL/6 mice: popular choice - DIO rate high (M)
Standard chow
10 kcal % fat
1-6 weeks
DIO: HFD
5-7 weeks
Wean
HFD on study
30-60% kcal fat
2-20 weeks
HFD 30-60
kcal % fat
1-2 weeks
Global (L) vs high socio-economic
index (R) macronutrient energy
composition, 2013
Schmidhuber et al. Lancet Planet Health 2018;2: e353–68
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14. Diet-Induced Obesity: High Fat
14
• C57BL/6J mouse is
a “gold standard”
Attribute C57BL6/J Mice Sprague-Dawley Rats
DIO proneness High, in males Intermediate, both sexes
Controls
Low % of DR
Littermates on standard chow
Abundant DR
Littermates on standard chow
Adiposity, body weight gain High, high High, mild
Hyperglycemia Mild Mild
Insulin resistance Yes Yes
Hypercholesterolemia Mild Yes
Hyperglyceridemia Yes Yes
Beta cell pathology No No
NAFLD (steatosis) Mild Yes
NASH No No
https://www.jax.org/jax-mice-and-
services/strain-data-sheet-pages/
phenotype-information-380050
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15. DIO Challenges and Solutions
15
Challenge Impact Solution
Induction time
6 weeks for metabolic syndrome
20+ weeks for severe adiposity
Commercial off-the shelf obese animals
Accelerate morbidities animal fats
Study-to-study variabilities:
• Fat content (30-60% kcal)
• Fat source (lard, vegetable, fish)
• Micronutrient and protein content
Reproducibility of methods, data:
• Energy intake
• Body composition
• Weight gain
• Study duration
• Comorbidities and pathologies
Defined, standardized research diets
(D12492, D12451)
Pair composition of HFD and control chow:
only significant change is fat content
Variable responses at strain or
substrain level
• C57BL/6J vs C57BL/6N vs
C57BL/6Ntac
• Sprague-Dawley from different
breeding facilities
Pre-sort DIO from DR early into HFD feeding
Minimize handling/transit stress
Induction time
6 weeks for metabolic syndrome
20+ weeks for severe adiposity
Commercial off-the shelf obese animals
Accelerate morbidities animal fats
Lack of salient comorbidities
Not suitable for NASH
Not suitable for advanced diabetes
Westernize diet composition
Genetic manipulations
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16. Diet Considerations:
Western-Style for NAFLD/NASH
16
• In addition to quantity, carbohydrate or fat quality impact
obesity and comorbidities, especially liver disease
• Increased fructose consumption via sucrose and corn
syrup is linked to the human obesity crisis
– Does not induce insulin and leptin secretion (reduced satiety)
– Induces lipogenesis and hepatic triglyceride synthesis
• Trans-fat, saturated fat, and long-chain fats promote
hepatic triglyceride accumulation and inflammation
– Decrease HDL, increase LDL
– Increase inflammation
– Impair hepatic insulin sensitivity
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17. 17
• C57BL/6J mice on a high fat diet
with fructose and glucose
accelerate toward NAFLD/NASH
– 60% kcal fat
– 2.31% w/vol fructose, 1.89% w/vol
glucose in drinking water
Liu et al. Lab Invest 2018;98: 1184-99
Western-Style for NAFLD/NASH
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18. 18
Liu et al. Lab Invest 2018;98: 1184-99
Western-Style for NAFLD/NASH
• C57BL/6J mice on a high fat
diet with fructose and
glucose accelerate toward
NAFLD/NASH
– 60% kcal fat
– 2.31% w/vol fructose, 1.89%
w/vol glucose in drinking water
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19. 19
Hansen et al. BMC Gastroenterol 2020;2: 210
C57BL/6J mice on GAN for 38 weeks
Alternative Diet Considerations
• AMLN/GAN: next
generation of
translatable
NASH diets
– 40% kcal fat, w/w:
22% trans + 26%
saturated (AMLN)
46% saturated (GAN)
– 22% w/w fructose
– 10% w/w sucrose
– 2% w/w cholesterol
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20. 20
• Bypass longer
induction time with
off-the-shelf induced
animals from
commercial vendors
• Fibrosis is key
therapeutic
endpoint, and
progresses with diet
duration
NASH Phenotype
www.taconic.com/mouse-model/diet-induced-nash-b6
H&E Picrosirius Red
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21. Corporate Headquarters:
16550 West Bernardo Drive
Building 5, Suite 525
San Diego, CA 92127
Tel: 855.827.6968
Fax: 888.882.4881
www.crownbio.com
Genetic Obesity Models

22. Leptin Knockout Animals:
Severe Obesity and Overt T2D
22
• Standard and advanced DIO models are useful for
obesity, prediabetic metabolic syndrome, and
NAFLD/NASH
• Overt diabetes is not a hallmark
• Spontaneous mutants in mouse colonies: hyperphagic,
obese, diabetic
• Enabled discovery of leptin: “the satiety hormone”
– Peptide hormone, key player along brain-adipose axis
– Synthesized by white adipocytes, proportionally to stored
triglyceride
– Targets hypothalamus; promotes satiety, stimulates metabolic rate
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23. Leptin Knockout Animals:
Severe Obesity and Overt T2D
23
• ob/ob affects leptin synthesis
– Commonly studied in C57BL/6J
background
• db/db affects leptin receptor
– C57BL/6J: moderate diabetes
– C57BKS: severe diabetes
• Obesity from hyperphagia, fat
cell hypertrophy, decreased
energy expenditure
• Leptin resistance is common in
obesity but leptin deficiency is
extremely rare cause of human
obesity https://www.jax.org/strain/000697https://www.jax.org/strain/000632
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24. Leptin Knockout Animals:
Severe Obesity and Overt T2D
24
• Leptin insensitive rats are available in
the Zucker fatty
• fa- allele maps to the leptin receptor
(analogous to db)
• Insulin resistant but normoglycemic
• Selective inbreeding for hyperglycemia
led to the
diabetic fatty (ZDF) with
severe onset type 2 diabetes
https://www.criver.com/products-services/find-model/zucker-rat
Shiota & Printz. Methods Mol Biol 2012;933: 103-23
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25. 25
• Heterozygous for ectopically
expressed allele of Agouti (signaling
peptide)
• Powerful, widespread antagonism of
MCR3 and MCR4 receptor in
hypothalamic neurons
• Adiposity due to hyperphagia,
adipose proliferation, adipose
hypertrophy
• 1-2.5% of human obesity associated
with MCR4 mutations
https://www.jax.org/strain/002468
https://www.jax.org/strain/002468
Agouti Yellow Mice:
Obesity and Overt T2D
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26. 26
Attribute
ob/ob (B6)
(JAX000632)
db/db (B6)
(JAX000697)
db/db (BKS)
(JAX000642)
KK-Ay/a (KK.Cg)
(JAX002468)
Genotype
Monogenic mutant
Leptin hormone
Monogenic mutant
Leptin receptor
Monogenic mutant
Leptin receptor
Monogenic mutant
Agouti overexpressor
Controls Wild type or ob/+ Wild type or db/+ Wild type or db/+ Non-agouti (KK-a/a)
Obesity, adiposity Severe (>40%) Severe (>40%) Moderate (>30%) High (>35%)
Basal
hyperglycemia
Moderate, remissive with
aging
Moderate, remissive with
aging
Severe, sustained
Moderate progressing to
severe (moderate,
sustained in KK-a/a)
Insulin resistance Severe, rapid Severe, rapid Severe, rapid
Severe, rapid (remissive
in KK-a/a with aging)
Hyperinsulinemia Yes Yes
Yes, rapidly becoming
hypoinsulinemia
Yes, later becoming
hypoinsulinemia
Hyper-
cholesterolemia
Yes Yes Yes Yes
Hyperlipidemia Mild Mild Yes Severe
Beta cell changes Hyperplasia, hypertrophy Hyperplasia Hypertrophy then atrophy Hyperplasia, hypertrophy
Preclinical utility
Obesity, Met-S, T2D,
steatosis
Obesity, Met-S, T2D,
steatosis
Obesity, Met-S, T2D, T1D,
NAFLD
Obesity, Met-S, T2D, T1D,
steatosis
Monogenic Mice At-A-Glance
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27. 27
Attribute Zucker fatty ZDF
Genotype
Monogenic mutant
(leptin receptor)
Monogenic mutant (leptin receptor)
Monogenic mutant? (beta cell transcription)
Controls Zucker wild type or fa/+ Zucker wild type or fa/+
Obesity, adiposity Severe (40-50%) Mild (>30%)
Basal hyperglycemia No Severe after onset
Insulin resistance Yes Severe, rapid
Hyperinsulinemia Yes
Yes, early
Hypoinsulinemia later
Hypercholesterolemia Yes Yes
Hyperlipidemia Yes Yes
Beta cell changes Hypertrophy Early failure
NAFLD Yes No
NASH Yes, on HFD No
Preclinical utility Obesity, Met-S, steatosis, T2D T2D, T1D, advanced diabetic comorbidities
Monogenic Rats At-A-Glance
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28. How Translational are
Monogenic Models?
28
• Human obesity, diabetes, NAFLD/NASH have polygenic etiologies
• Mutation in the leptin pathway results in a powerful phenotype.
– Very rare in humans
• Antagonized melanocortin receptor pathways results in atypical tissue
proliferation.
– Somewhat rare in humans, phenotype is more subtle
• Animals selectively bred for polygenic origin of obesity proneness better
capture the human condition
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29. 29
Polygenic Models: Obesity with
Relevant Comorbidities
The MS-NASH mouse has intact
leptin signaling in contrast to
monogenic obesity
C57BL/6
DIO
MS-NASH
C57BL/6 (Standard Diet)
DIO (HFD)
MS-NASH (Standard Diet)
MS-NASH
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30. 30
MS-NASH Model Disease Phenotype
MS-NASH is:
• Hyperinsulinemic
• Hyperglycemic
• Hypertriglyceridemic
• On a standard diet
• Shows impaired
glucose tolerance
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31. 31
Disease Progression in MS-NASH
Animal Age (in weeks)8 32
24 weeks on Western Diet + 5% Fructose*
0 4 8 12 2416 20
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32. 32
Liver Disease Phenotype
e k s o n D ie t
W e e k s o n D ie t
8 1 2 1 6 2 0
e e k s o n D ie t
***
*** ***
***
0 4 8 1 2 1 6 2 0
0
1 0 0
2 0 0
3 0 0
4 0 0
5 0 0
W e e k s o n D ie t
AST(U/L)
***
***
*** ***
*
8 1 2 1 6 2 0
W e e k s o n D ie t
*
* *
*
0 4 8 1 2 1 6 2 0
0
1 0 0
2 0 0
3 0 0
W e e k s o n D ie t
Triglycerides(mg/dL)
*
* * *
8 1 2 1 6 2 0
e e k s o n D ie t
*
* * *
1 2 1 6 2 0
0
2 0
4 0
6 0
W e e k s o n d ie t
LiverTG
(mg/gtissue)
*
* *
0 4 8 1 2 1 6 2 0
0
1 0 0
2 0 0
3 0 0
4 0 0
W e e k s o n D ie t
ALT(U/L)
***
*** ***
***
0 4 8 1 2 1 6 2 0
0
1 0 0
2 0 0
3 0 0
4 0 0
5 0 0
W e e k s o n D ie t
AST(U/L)
***
***
*** ***
*
0 4 8 1 2 1 6 2 0
0
5
1 0
1 5
W e e k s o n D ie t
LiverWeight(%BW)
*
* * *
*
1 2 1 6 2 0
0
2 0
4 0
6 0
W e e k s o n d ie t
LiverTG
(mg/gtissue)
*
* *
0 4 8 1 2 1 6 2 0
2 0
3 0
4 0
5 0
W e e k s o n D ie t
BodyWeight(g)
* *
W D F (n = 8 )
C D (n = 8 )
0 4 8 1 2 1 6 2 0
2 0
3 0
4 0
W e e k s o n D ie t
%BodyFat
*
* * *
2 0 0
3 0 0
4 0 0
LT(U/L)
***
*** ***
***
2 0 0
3 0 0
4 0 0
5 0 0
ST(U/L)
***
***
*** ***
0 4 8 1 2 1 6 2 0
0
1 0 0
2 0 0
3 0 0
4 0 0
W e e k s o n D ie t
TotalCholesterol
(mg/dL)
* *
* *
*
0 4 8 1 2 1 6 2 0
0
1 0 0
2 0 0
3 0 0
W e e k s o n D ie t
Triglycerides(mg/dL)
*
* * *
1 6 2 0
0
1
2
3
W e e k s o n D ie t
SteatosisScore(0-3)
*
C D (n = 8 )
W D F (n = 8 )
1 6 2 0
0 .0
0 .5
1 .0
1 .5
2 .0
W e e k s o n D ie t
BallooningScore(0-2)
*
*
1 6 2 0
0
1
2
3
W e e k s o n D ie tLobularInflammationScore(0-3)
* *
1 6 2 0
0 .0
0 .5
1 .0
1 .5
2 .0
W e e k s o n D ie t
FibrosisScore(0-4)
*
*
1 6 2 0
0
2
4
6
W e e k s o n D ie t
NAFLDActivityScore
* *
H&E
PSR
0 4 8 1 2 1 6 2 0
2 0
3 0
4 0
5 0
W e e k s o n D ie t
BodyWeight(g)
* *
W D F (n = 8 )
C D (n = 8 )
0 4 8 1 2 1 6 2 0
2 0
3 0
4 0
W e e k s o n D ie t
%BodyFat
*
* * *
0 4 8 1 2 1 6 2 0
0
1 0 0
2 0 0
3 0 0
4 0 0
W e e k s o n D ie t
ALT(U/L)
***
*** ***
***
0 4 8 1 2 1 6 2 0
0
1 0 0
2 0 0
3 0 0
4 0 0
5 0 0
W e e k s o n D ie t
AST(U/L)
***
***
*** ***
*
0 4 8 1 2 1 6 2 0
0
1 0 0
2 0 0
3 0 0
4 0 0
W e e k s o n D ie t
TotalCholesterol
(mg/dL)
* *
* *
*
0 4 8 1 2 1 6 2 0
0
1 0 0
2 0 0
3 0 0
W e e k s o n D ie t
Triglycerides(mg/dL)
*
* * *
1 0
1 5
ight(%BW)
*
* * *
*
4 0
6 0
erTG
tissue)
*
* *
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33. 33
ZDSD Metabolic Rat Model
The ZDSD rat has:
• Intact leptin signaling
• Obese on a standard diet
Selective breeding for obesity and diabetes
In-breeding (35+ generations)
Crl:CD(SD) ratZDF rat (Lean+/+)
ZDSD
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34. 34
Spontaneous Development of
Diabetes in ZDSD
Age (years)
50 6035
Age (weeks)
Human
ZDSD Rat
Age (years)
50 6035
Age (weeks)
• ZDSD is spontaneously diabetic
• Insulin resistance and decrease are similar to human patients
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35. 35
Diabetic Neuropathy in ZDSD Rats
Response to Standard
of Care
Thickening of Basement
Membrane
• ZDSD hallmarks of diabetic nephropathy
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36. Corporate Headquarters:
16550 West Bernardo Drive
Building 5, Suite 525
San Diego, CA 92127
Tel: 855.827.6968
Fax: 888.882.4881
www.crownbio.com
Additional Considerations for
Optimal Study Design

37. Additional Considerations
• Obesity is often considered a disease of excess
caloric intake, not altered caloric expenditure
• Physical activity, and food choice or feeding
behavior, are disregarded
37
• Automatable electronic monitoring and
feeding systems provide additional data with
reduced technician burden and animal
disruption
http://www.colinst.com/products/clams-
comprehensive-lab-animal-monitoring-system
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38. Additional Considerations
38
Automated food delivery
(time of day, quantity)
Running wheel measures
voluntary physical activity
Mass flow
measurements for O2
consumption and
CO2 emission
Implantable telemetry
devices for heart rate,
body temperature
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39. Additional Considerations
• Human Ta: 20-22ºC
– Humans expend little energy to
maintain Tb
– Mice expend >1/3 of energy to
maintain Tb, largely via
uncoupled fatty acid oxidation
in brown adipose tissue
• Mouse Ta: 30-32ºC
39
Abreu-Vieira et al. Mol Metab 2015;4: 461-470
Cannon & Nedergard.
J Exp Biol 2011;
214(Pt2): 242-253
• Thermoneutrality i.e. “ambient temperature”:
– Minimal dedication of energy toward heat generation
to maintain body temperature
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40. Additional Considerations
• Thermoneutrality affects mice profoundly
• Case study: High fat diet NAFLD pathogenesis is exacerbated at thermoneutrality
40
Increased body weight
Decreased food intake
Decreased arterial
pressure, heart rate
Decreased catecholamine,
corticosteroid production
Decreased thermogenic
energy expenditure
Enhanced
NAFLD/NASH
Giles et al. Nat Med 2017;23: 829-840
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41. Additional Considerations
41
Xiao et al. Obesity 2015;23: 1450-59
• Thermoneutrality affects mice profoundly
• Case study: thermoneutrality is required for pharmacological benefits of
β-andrenergic receptor agonist
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42. Summary
42
• High fat diet models provide a relatively easy route to obesity
– Weight gain/adiposity
– Steatosis
– Metabolic syndrome
• Adjuncts fructose, saturated/trans-fats, cholesterol adapt models for NASH
– More translational % of calories from fat
– Some characteristics of metabolic syndrome, hyperglycemia may not be one
• Leptin/MCR signaling mutants
– Rapid path to obesity, type 2 diabetes
– May lack translatability
• Polygenic mutants
– Many modalities for studying obesity, diabetes, and NASH
– Potentially more translationally relevant
• Technological, environmental upgrades to husbandry and monitoring add translatability to
studies on energy expenditure
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43. Contact CrownBio
43
Get in Touch!
• Email busdev@crownbio.com or contact your local Account Manager to:
– Obtain more information about our models
– Discuss this data with a scientific resource
– Initiate a new study
More Information
• Visit www.crownbio.com to learn more about CrownBio’s comprehensive
Translational Metabolic Disease Platforms
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44. Crown Bioscience Inc. │ Confidential – Not For Distribution

45. Fred Beasley, PhD
Director of Scientific Engagement,
Cardiovascular and Metabolic Disease
Crown Bioscience
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