Regulation of Systemic Energy Balance and Glucose...
Transcript of Regulation of Systemic Energy Balance and Glucose...
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Regulation of Systemic Energy Balance and Glucose Homeostasis by Protein-Tyrosine
Phosphatase 1B
Fawaz G. Haj
Associate Professor
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Diagnosed Obesity and Diabetes for Adults aged ≥ 20 years in United States
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The Obesity Epidemic Has Reached America’s Pets
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Increased hepaticglucose production
Muscle
GlucoseImpaired glucose
utilization
AdipocytesInsulin secretorydefect
Insulin
Liver
Pancreas
+
System Regulation of Glucose
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Understand the Role of Cell Signaling, in Particular, Tyrosine Phosphorylation in the
Pathogenesis of Metabolic Diseases
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Phosphorylation is reversible
Y Y Y Y
Y YProteinProtein
PPPP
ProteinProtein
PP
PPPP
PTPsPTPs
PTKsPTKs
PP
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Andersen et al. 2001 http://ptp.cshl.edu
Nontransmembrane PTP subtypes (NT) Receptor-like PTP Subtypes (R)
The Classical Protein-Tyrosine Phosphatase (PTP) Family
HCSxGxGRxG
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WT
KO
• PTP1B KO mice are lean and resistant to diet-induced obesity (Elchebly et al, 1999 andKlaman et al, 2000)
Protein-Tyrosine Phosphatase 1B (PTP1B)
• PTP1B KO mice are insulin-sensitive, and have increased insulin receptor (IR) phosphorylation (Elchebly et al, 1999 and Klaman et al, 2000)
Is ubiquitously expressed, including all insulin-responsive tissues
Generate PTP1B knockout (KO) mice (protein no longer expressed)
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PTP1B Inhibition for Treatment of Metabolic Diseases
No obvious apparent side effects observed thus far, making PTP1B a good target for treating obesity and diabetes in humans
Companies are competing to generate a readily bio-available, PTP1B-specific inhibitor
Need to know the SITES and MECHANISMSof PTP1B action
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Genetically-engineered mice to achieve tissue-specific deletion of PTP1B
Quantitative cellular imaging to dissect the mechanism of PTP1B action
Approach
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Genetically-engineered mice to achieve tissue-specific deletion of PTP1B
Quantitative cellular imaging to dissect the mechanism of PTP1B action
Approach
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PTP1B fl/fl
PDX1-Cre
Alb-Cre
Adp-Cre
Pancreas
Liver
Adipose
Tissue-Specific Deletion of PTP1B
MCK-Cre Muscle
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PTP1B fl/fl Adipoq-Cre
PTP1B adipose-specific deletion
Generation of Adipose-Specific PTP1B Knockout Mice using Adipoq-Cre
X
FPTP1B KO
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W1 CW1 W1 CW1 W2 CW2 W1 CW1 W2 CW2 W3 CW3
Adipoq WT KO
PTP1B
PTP1
B e
xpre
ssio
n
ERK1/2
WCW
*
##
0
25
50
75
100
125
Adipoq WT KO
W CW BAT Mac L M P Br W CW BAT Mac L M P Br
TCPTP
SHP2
PTP1B
ERK1/2
WT KO
Efficient and Specific Deletion of PTP1B in Adipose Tissue
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Age (weeks) Age (weeks)
Bod
y w
eigh
t (g)
Age (weeks)
Bod
y w
eigh
t (g)
Age (weeks)
Bod
y w
eigh
t (g)
Bod
y w
eigh
t (g)
# #***# *# #**#**
Females/HFD
Males/Chow Females/Chow
Males/HFD
# #** # #** # #** # #**# #**
0
10
20
30
40
50
3 5 7 9 11 13 15 17 19 21 23
0
10
20
30
40
50
3 5 7 9 11 13 15 17 19 21 23
0
10
20
30
40
50
3 5 7 9 11 13 15 17 19 21 23
0
10
20
30
40
50
3 5 7 9 11 13 15 17 19 21 23
Resistance to HFD-Induced Obesity in adipose PTP1B KO Mice
flx/flx
flx/flx, Cre (KO)+/+, Cre
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flx/flx
+/+, Cre (Adipoq)flx/flx, Cre (KO)
Ener
gy E
xpen
ditu
re (K
J/M
in/K
g0.
75)
** **
*
0
200
400
600
800
1000
1200
Dark Fed Light Fed Dark Fast Total
Increased Energy Expenditure in Adipose PTP1B KO Mice
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Genetically-engineered mice to achieve tissue-specific deletion of PTP1B
Quantitative cellular imaging to dissect the mechanism of PTP1B action
Approach
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Fluo
resc
ence
inte
nsity
Time
Pre-bleach
BleachRecovery
Fluorescence Recovery After Photobleaching (FRAP)
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Dynamics of Cellular PTP1B Mobility
PTP1B WT (ER) PTP1B D/A (ER) PTP1B D/A (Cell-Cell)
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Quantitative Determination of PTP1B Mobility Using FRAP
Recovery consistent with: 1) Rapid turnover (<10 sec)2) Replenishment from ER pool 3) No significant “tight binding” fraction
D/A cell-cell
(Deff: Effective diffusion)
WT ER, Deff = 0.20 µm2 s-1
D/A ER, Deff = 0.07 µm2 s-1
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Summary
Adipose-specific PTP1B deletion leads to decreased adiposity and resistance to high fat diet-induced obesity
This is due to increased energy expenditure in adipose PTP1B deficient mice
Cellular imaging identifies the endoplasmic reticulum and cell-cell contact as major cites of PTP1B action in the cell
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Conclusion
Tissue-specific deletion and cell biology approaches are necessary to decipher the sites and mechanisms of
PTP1B action
This is important for developing drugs that targets PTP1B at the correct site(s)
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Future Studies
Investigate the role of PTP1B in other tissues and it mechanism of action at these cites
Investigate the role of other members of PTP superfamily as potential targets to treat obesity and
diabetes
Examine regulation of PTP activity by nutrients and diets
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Acknowledgements
Collaborators
Peter Havel Lab (UCD)
Lew Cantley Lab (Harvard)
Philippe Bastiaens Lab (MPI)
Thanks
Ben NeelEvan Rosen
Financial support from JDRF, ADA and NIH
FGH Lab
Ahmed BettaiebJesse BakkeSiming LiuYannan XiNaoto NagataDanny AbouBechara
Kosuke MatsuoIzumi Matsuo