Dr. Mehmet Furkan Burak

ACP Annual Scientific Meeting, MA 2016

Development of anti-aP2 therapeutics against diabetes and fatty liver disease

Financial Disclosure

I don’t have any conflict of interest

Rosen, Cell 2014

70 Trillion $

Obesity recognized as a disease

Spiegelman, Nature 2006

aP2

Adipocyte Fatty Acid Binding Protein (FABP4-aP2)

Furuhashi, Nature Rev 2008

Genetic deficiency of aP2 is protective in many diseases

aP2

aP2-/-aP2dele on

aP2 & Diabetes

aP2

Deficiency of adipocyte FABPs protects against a

variety of metabolic diseases

Boord et al 2004

Maeda et al 2005

Makowski et al 2002 FABP-/- = dual aP2/FABP5 KO

Shum et al 2006

Evidence that aP2 is important in metabolic disease

Hotamisligil Confidential

Blood

Gluco

se (mg/

dl)

Maeda2005CellMetabolism

aP2 & Atherosclerosis

aP2

Deficiency of adipocyte FABPs protects against a

variety of metabolic diseases

Boord et al 2004

Maeda et al 2005

Makowski et al 2002 FABP-/- = dual aP2/FABP5 KO

Shum et al 2006

Evidence that aP2 is important in metabolic disease

Hotamisligil Confidential

Ath

ero

sclero

sis

+/+ -/-

Deficiency of adipocyte FABPs protects against a

variety of metabolic diseases

Boord et al 2004

Maeda et al 2005

Makowski et al 2002 FABP-/- = dual aP2/FABP5 KO

Shum et al 2006

Evidence that aP2 is important in metabolic disease

Hotamisligil Confidential

Boord2004Circula on

aP2 & Fatty Liver Disease

aP2

Deficiency of adipocyte FABPs protects against a

variety of metabolic diseases

Boord et al 2004

Maeda et al 2005

Makowski et al 2002 FABP-/- = dual aP2/FABP5 KO

Shum et al 2006

Evidence that aP2 is important in metabolic disease

Hotamisligil Confidential

Hepato

steato

sis

aP2 ?

Serum aP2 levels increased with obesity Serum aP2 increases with BMI and obesity

7

Serum aP2 in genetic obesity in

mouse and man

Serum aP2 and BMI

Cao & Hotamisligil, unpublished

Data suggest mouse may be a good model for human

Serum aP2 increases with BMI and obesity

Hotamisligil Confidential

0

0

0

0

0

0

Fast

ing

glu

cose

an

d in

sulin

Promoter polymorphism results in decreased adipose aP2 expression n: 7899

0

2

4

6

8

10

12

14

16

18

1 2

0

2

4

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12

14

1 2

0

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4

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1 2

0

5

10

15

20

25

1 2

GIR

(m

g/k

g/m

in)

RD

(m

g/k

g/m

in)

*

C-H

GP

(m

g/k

g/m

in)

B-H

GP

(m

g/k

g/m

in)

PI-IgG aP2 antibody

*

*

PI-IgG aP2 antibody PI-IgG aP2 antibody

PI-lgG aP2 antibody

A B C

D

Figure 6

E

Lipolysis

β-adrenergic stimulation

MVB

Other

Organs ?

Glucose Production

aP2

aP2

Treatment & tests HFD – become obese 6 21 25

Birth

Week 0

HFD

Vehicle or anti-aP2 monoclonal antibody 33mg/kg, 2x/week

subQ injections

Study Design

- Weekly Fasting Blood glucose

Fed Body Weight

- Week 2 Glucose tolerance test

- Week 3 Insulin tolerance test

- Week 4-5 Clamp – SAC

- Histological and molecular tests

from week 4-5 tissues.

Give me the same!

Sorry! You are

in control group

The monoclonal anti-aP2 antibody CA33 improved glucose metabolism

0

2

4

6

8

10

PBS CA33 CA13 CA15 CA23 H3

Insulin(n

g/ml)

*

*

* *

*

n.s

Vehicle

CA33

CA13

CA15

CA23

HA3

Week0 Week4

0

50

100

150

200

PBS CA33 CA13 CA15 CA23 H3

Glucose(mg/dl)

**

CA33

CA13

CA15

CA23

HA3

Vehicle

*

Week0 Week4

0

100

200

300

400

0 30 60 90 120

Vehicle

CA33

Timea erglucose(min)

Glucose(mg/dL)

**

Glucose(mg/dL)

Timea erinsulin(min)

0

50

100

150

200

0 20 40 60 80 100

Vehicle

CA33

**

A. B.

C. D.

Burak et al. Science TM 2015

The monoclonal anti-aP2 antibody CA33 improved glucose metabolism

0

50

100

150

200

250

Week 0 Week 1 Week 3 Week 5

PBS CA33

Glu

cose

(m

g/d

l) *

** **

6 hr fasting

Burak et al. Science TM 2015

C

0

100

200

300

400

0 30 60 90 120

PBS

CA33

Glucose(mg/dl)

Timea erglucose(min)

HFD-aP2-/-

Vehicle

A B

Bodyweight(g)

Week0 Week3

HFD-aP2-/-

0

20

40

60

80

100

120

140

160

1 2Vehicle CA33

HFD-aP2-/-

Glucose(mg/dL)

Week0 Week3

0

10

20

30

40

50

1 2Vehicle CA33

E

0

100

200

300

400

500

600

700

0 30 60 90 120

VehicleCA33

Timea erglucose(min)

Glucose(mg/dl)

ob/ob

*

D

0

100

200

300

400

500

1 2Vehicle CA33

Glucose(mg/dL) **

Week0 Week3

F

0

2

4

6

8

10

12

PBS CA33

Insulin(ng/ml)

**

Vehicle CA33

CA33 effect is specific to aP2 and robust in different obesity models

Burak et al. Science TM 2015

PEPCK Pck1

G6pc

SAC

clamp Steady state basal acclimatization

Tissue collection

-5 0 5 45 10 15...

2 hr 2.5 hr 45 min

Clamp Settings

+++ Insulin + Glucose H-glucose C-glucose 3

14

CA33 decreased liver glucose production and increased peripheral insulin sensitivity

Burak et al. Science TM 2015

1 cm 1 cm

PBS CA33

0

1

2

3

4

5

6

Liver Weight(g)

Liver % Bodyweight

PBS

CA33

**

**

*

*

CA33 treatment improved lipid metabolism

and reversed hepatosteatosis in obese mice

Burak et al. Science TM 2015

0

50

100

150

200

250

PBSCA33

Cholesterol(mg/dl)

VehicleCA33

*

mRNA/36B4

**

**

0

0.5

1

1.5

Scd1 Fasn Acc1

**

Scd1 Fasn Acc1

Vehicle CA33C D

CA33 treatment improved lipid metabolism

and reversed hepatosteatosis in obese mice

Burak et al. Science TM 2015

0

5

10

15

20

25

30

35

40

45

50

0 1 2 3 4 5

PBS

CA33

Bo

dy

We

igh

ts (

g)

* * * *

CA33 prevented HFD induced weight gain and decreased BW

weeks

CA33 decreased fat mass

B

PGWATWeight(g)

0

10

20

30

40

50

60

70

PBS CA33

Fatmass(g)

Leanmass(g)

Vehicle CA33

*

Bod

ycompo

sion

0

1

2

3

4

5

6

LiverWeight %Bodyweight

**

**

LiverWeight

%BodyWeight

Weight

Vehicle

CA33

DCA

0

0.5

1

1.5

2

2.5

Vehicle CA33VehicleCA330

0.1

0.2

0.3

0.4

0.5

BATWeight(g)

VehicleCA33

**

**

E

Vehicle CA33

Burak et al. Science TM 2015

0

1

2

3

4

5

6

7

8

9

PBS CA33

Totalfoodintake(g)

n.s.

VehicleCA33

Acvitycounts(AU)

n.s.

0

100

200

300

400

500

Light Dark

n.s.Vehicle

CA33

Light Dark0.70

0.75

0.80

Light Dark

PBS CA33

**

0

500

1,000

1,500

2,000

2,500

3,000

3,500

Light Dark

PBS CA33**

Light DarkVO2(ml/kg/hr)

RER

Light Dark

**

**

E F G H

CA33 accelerated oxidative metabolism and decreased fat mass

Burak et al. Science TM 2015

- aP2 secretion increased with fasting.

- Serum aP2 levels increased with human obesity and contributes to hyperglycemia.

- Secreted aP2 can be effectively targeted by a monoclonal antibody to ameliorate diabetes and reduce fat mass and hepatic steatosis.

- CA33, as a potential preclinical molecule that lowered fasting blood glucose, improved glucose metabolism, increased systemic insulin sensitivity and reduced liver steatosis.

- These metabolic outcomes were; -reproducible in two independent models of obesity, genetic and dietary, -specific to aP2, -linked to the regulation of hepatic glucose output and peripheral glucose utilization.

- CA33 also decreased fat mass and increased oxidative capacity without any effect on physical

activity and food intake.

Summary

Acknowledgement

DP2 transition aP2

8 November 2013 Dr. Gökhan Hotamisligil

Dr.Valerie Stone

Dr.Patrick Gordan Dr.Barbara Blair