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GAPDH
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The National Mouse Metabolic Phenotyping Center (MMPC) @ UMass is designed to
provide the scientific community with sophisticated and standardized experimental
tools for the purpose of investigating transgenic mouse models and understanding
obesity, diabetes and its complications. The MMPC @ UMass is composed of a
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assess insulin sensitivity, glucose/lipid/protein metabolism, body composition and
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throughput measurement of serum/tissue hormones, metabolites and cytokines known
to affect metabolism.
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The 2016 NIH Summer Research Fellowship Program at UMass Medical School
NIH Grant #2 R25 HL092610-07 Website: http://www.umassmed.edu/summer
Transgenic Mouse Model of Breast Cancer Causes Skeletal Muscle
Inflammation and Insulin Resistance
Stephanie Choi1, Hee Joon Kang1, Suchaorn Saengnipanthkul1, Kunikazu Inashima1, Hye-­Lim Noh1,
Randall H. Friedline1, Jose Mercao-­Matos3, Leslie M. Shaw3 and Jason K. Kim1,2
1Program in Molecular Medicine, 2Department of Medicine, Division of Endocrinology, Metabolism and Diabetes, and 3Department of Cell & Cancer Biology
Stephanie.Choi@umassmed.edu
[1] Cairns, R. et al. (2011) Regulation of cancer cell metabolism. Nature
Reviews Cancer, 11: 85-­95.
[2] Cartensen, B. et al. (2016). Cancer incidence in persons with type 1
diabetes: a five-­country study of 9,000 cancers in type 1 diabetic
individuals. Diabetologia. 59(5): 980-­988.
[3] Gray, S. & Kim, J. (2011). New insights into insulin resistance in the diabetic
heart. Trends in Endocrinology & Metabolism, 22: 389-­428.
[4] Janowska J. et. al. (2006) Relationship between serum resistin
concentration and proinflammatory cytokines in obese women with
impaired and normal glucose tolerance. Metabolism. 55 (11): 1495-­1499.
[5] Kang, Y.E. et. al. (2016). The roles of adipokines, proinflammatory cytokines,
and adipose tissue macrophages in obesity-­associated insulin resistance
in modest obesity and early metabolic dysfunction. PLos One. 11(4).
[6] Larsson, S.C. et al. (2007). Diabetes mellitus and risk of breast cancer: A
meta-­analysis. Int. J. Cancer. 121: 856-­862.
[7] Peake, J. et al. (2015). Cytokine expression and secretion by skeletal
muscle cells: regulatory mechanisms and exercise effects. 21: 8-­25.
[8] United States Cancer Statistics: 1999-­2012 Incidence and Mortality Web-­
based Report. U.S. Cancer Statistics Working Group. Atlanta, GA: U.S.
Department of Health and Human Services, Centers for Disease Control
and Prevention, and National Cancer Institute.
Our findings suggest a novel paradigm in which
inflammatory cytokines and macrophages derived from
the tumor microenvironment affects muscle glucose
metabolism and causes muscle insulin resistance.
Epidemiological evidence has highlighted a relationship
between breast cancer and diabetes. Furthermore,
breast cancer is associated with an inflammatory
response, but the underlying mechanisms leading to
inflammation and its effect on glucose metabolism and
insulin signaling remain poorly understood. Here we
examined glucose metabolism in a transgenic mouse
model of breast cancer expressing the polyoma middle
T-­antigen oncogene driven by the Mouse Mammary
Tumor Virus promoter (MMTY-­PyMT). A
hyperinsulinemic-­euglycemic clamp was performed in
female MMTV-­PyMT and WT mice at 8-­9 weeks of age.
Whole body glucose turnover rates were significantly
decreased in MMTV-­PyMT mice with a 20% decrease
in insulin-­stimulated glucose uptake observed in
skeletal muscle. More macrophages were found in
skeletal muscle of MMTV-­PyMT mice, as suggested by
increased CD68 and F4/80 mRNA levels.. Consistent
with this, MMTV-­PyMT mice also had a significant
increase in circulating plasma levels and skeletal
muscle mRNA expression of IL-­6, MCP-­1, and G-­CSF.
In particular, there was a trend of increased mRNA
expression of Arg1, a M2 macrophage-­specific marker.
A similar trend was observed in tumor tissues, but with
much higher gene expression of inflammatory
cytokines and Arg1 as compared to skeletal muscle.
Skeletal muscle of MMTV-­PyMT mice showed a trend
towards higher expression of the stress-­response
protein, CHOP, possibly caused by increased
inflammation. These results indicate that tumor-­bearing
MMTV-­PyMT mice develop insulin resistance in
skeletal muscle, along with increased gene expression
of macrophage markers and levels of inflammatory
cytokines. Taken together, our findings suggest a novel
paradigm in which tumor microenvironment-­derived
inflammatory cytokines affect muscle glucose
metabolism, providing new insights into the relationship
between breast cancer and insulin resistance.
Breast Cancer Mouse Model: MMTV-­PyMT+ Transgenic Mice
Ø MMTV-­PyMT mice that express Polyoma Virus middle T antigen (PyMT) under
the direction of the mouse mammary tumor virus (MMTV) promoter and WT mice
at 8~9 weeks of age (n=9~12 per group)
Ø A 2-­hr hyperinsulinemic-­euglycemic clamp with [3H]-­glucose and [14C]-­2-­DG was
conducted in awake mice to measure insulin sensitivity and glucose metabolism
Ø Western blot, RT-­qPCR, and multiplexed Luminex assay were used to assess
insulin signaling and inflammation in peripheral organs.WT PyMT+
Abstract
q Epidemiological evidence suggests an association
between both type 1 diabetes mellitus (T1DM) [2]
and type 2 diabetes mellitus (T2DM) with increased
breast cancer incidence and mortality [6,8]
q Insulin resistance, or impaired insulin sensitivity, is a
key characteristic of T2DM. It results in impaired
glucose metabolism [3,4]
q According to the Warburg effect, cancer tumor cells
exhibit shifting in ATP generation through oxidative
phosphorylation to using glycolysis, thus requires an
abnormally high rate of glucose uptake [1], which
may impact glucose metabolism in other tissues.
q Inflammation along with elevated levels of cytokines
is a major event during obesity-­mediated insulin
resistance [5,7]
Study Design Proposed Model
Error bars indicate standard error.*P < 0.05, **P < 0.01 mark degree of statistical significance between groups.
This study was funded by NIH grants (R01-­DK080756
and R24-­DK090963), and the National Mouse
Metabolic Phenotyping Center at UMass (U24-­
DK093000). I would like to extend a thank you for the
support and assistance offered by Mrs. Karen J.
Zirpola-­Miller, Dr. Deborah H. Hines, Dr. Brian Lewis,
Dr. Regino Mercado-­Lubo, and Mrs. Linhelle Charles.
To examine the inflammatory effects of breast cancer in
MMTV-­PyMT mice on insulin action and glucose
metabolism
Tumor
Tumor
microenvironment
Skeletal Muscle
Mac
Mac
Mac
Mac
Mac
G-­CSF
MCP-­1
IL-6
Insulin Resistance
Glucose metabolism
Cytokines
Macrophages
Mac
Future studies will evaluate the roles of specific
inflammatory cytokines and macrophages in regulating
systemic glucose metabolism and insulin signaling
pathway via inflammation in breast cancer tumor model.
*
*
QD Tumor
Whole Body Glucose
Turnover
Skeletal Muscle
Glucose Uptake
*
Reduced Glucose Metabolism and Insulin Sensitivity in PyMT+ Mice
Hepatic Insulin Action
IL-­6 G-­CSF MCP-­1
*
*
**
**
*
QD Tumor QD Tumor
WT PyMT WT PyMT
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Results
Introduction
Specific Aim
Conclusion
Future Studies
References
Acknowledgements
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CD11c Arg1 CD11c Arg1
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Tumor
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*
105
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p = 0.13
PyMT+ Mice Have Increased Macrophage Marker mRNA Expression
2
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Analysis of Stress Signaling in Skeletal Muscle of PyMT+ Mice
WT QD PyMT QD