Hannelouise Kissow

Department of Biomedical Sciences Faculty of Health and Medical Sciences

7/10/18 1

TGR5 activation induces intestinal growth and improves intestinal mucositis in mice

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Background GLP-1 and GLP-2 are secreted from L cells

•  Hormones are co-distributed •  Secreted after oral food intake •  Exogenous GLP-2 ameliorates GI mucositis

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Compensatory hyperproliferation

and healing

Background GLP-1 and GLP-2 are essential for intestinal healing

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Compensatory hyperproliferation

and healing

Background GLP-1 and GLP-2 are essential for intestinal healing

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0 1 2 3 4 5

-2 0

-1 5

-1 0

-5

0

5

% c

ha

ng

e i

n B

W

WT Saline

WT 5-FU

Tg 5-FU

Tg 5-FU GLP-2

Tg 5-FU GLP-1

Tg 5-FU Co-treatment

Rasmus Hytting-Andreasen et al. Endogenous glucagon-like peptide- 1 and 2 are essential for regeneration after acute intestinal injury in mice PLoS One 2018; 13(6): e0198046

Background GLP-1 and GLP-2 are essential for intestinal healing

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Compensatory hyperproliferation

and healing

TGR5

Background TGR5 is expressed on the L-cells

•  Bile acid receptor •  G protein coupled •  Widely distributed •  Activation à GLP-1 secretion •  Increase gallbladder filling

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Maruyama T et al. Identification of membrane-type receptor for bile acids (M-BAR). Biochem Biophys Res Commun. 2002 Nov 15;298(5):714–9

Production of cAMP in CHO-TGR5 transfected cells

LCA > CDCA DCA > CA

Background Secondary BA’s are the most potent TGR5 activators’

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Background Microbiota controls the dehydroxylation

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Compensatory hyperproliferation

and healing

TGR5

CDCA CA

LCA DCA

Background Does microbiota control intestinal healing?

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Method

Main endpoints •  Intestinal weight •  Morphology •  Tissue hormone concentration

30 mg/kg oral gavage x 2

Main endpoints •  Body weight •  Small intestinal weight •  Morphology •  Mucositis severity

Ex 1

Ex 2

TGR5 agonist

Vehicle

Sacrificed after 10 days of treatment

Sacrificed 72 h after 5-FU (400 mg/kg i.p)

TGR5 agonist

Vehicle TGR5 agonist

Vehicle

5-FU 5-FU

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Results TGR5 activation induced small intestinal growth

R O 5 5 2 7 2 3 9 V e h ic le0

1

2

3

4

5

S m a ll In te s tin e W e ig h t

% o

f B

W

****

D u o d e n u m J e ju n u m Ile u m0

2 0 0

4 0 0

6 0 0

8 0 0

Vil

lus

he

igh

t (µ

m)

* *

*

RO5527239

V e h ic le

D u o d e n u m J e ju n u m Ile u m0

2 0

4 0

6 0

8 0

1 0 0

Cry

pt

de

pth

(µ

m)

R O 5 5 2 7 2 3 9 V e h ic le0 .0

0 .5

1 .0

1 .5

C o lo n W e ig h t

% o

f B

W

Mean ± SEM n=8 *=p < 0.05, **=p < 0.01, **** =p < 0.0001

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Results TGR5 activation induced GLP-1 and GLP-2 secretion

R O 5 5 2 7 2 3 9 V e h ic le0

5 0

1 0 0

1 5 0

A c t iv e G L P -1 in S m a ll In te s tin e

pm

ol/

g

R O 5 5 2 7 2 3 9 V e h ic le0

1 0

2 0

3 0

4 0

5 0

A c tiv e G L P -2 in S m a ll In te s tin e

pm

ol/

g

R O 5 5 2 7 2 3 9 V e h ic le0

1 0 0

2 0 0

3 0 0

4 0 0

A c tiv e G L P -1 in C o lo n

pm

ol/

g

* *

R O 5 5 2 7 2 3 9 V e h ic le0

5 0

1 0 0

1 5 0

A c tiv e G L P -2 in C o lo n

pm

ol/

g* *

Mean ± SEM n=8 **=p < 0.01

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Results TGR5 activation ameliorates GI mucositis

D u o d e n u m J e ju n u m Ile u m0

2 0 0

4 0 0

6 0 0

8 0 0

V illu s h e ig h t

µm

5 -F U + R O 5 5 2 7 2 3 9

5 -F U + v e h ic le

R O 5 5 2 7 2 3 9

V e h ic le*_ _ _

D u o d e n u m J e ju n u m Ile u m0

5

1 0

1 5

M u c o s it is s e v e r ity s c o r e

sc

ore

5 -F U + R O 5 5 2 7 2 3 9

5 -F U + V e h ic le_ _ _ _ _

* _ _ _ _ _*

C h a n g e in B W

0 1 2 3-1 0

-5

0

5

1 0

%

Mean ± SEM n(5-FU)=8, n(control)=4 *=p < 0.05

Conclusion

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Activation of the BA receptor TGR5 led to small intestinal growth and GLP-1 and GLP-2 secretion in the colon Activation of the BA receptor TGR5 was able to ameliorate 5-FU induced GI mucositis Perspective

s

Financial support A. P. Møller Foundation Krista and Viggo Petersen Foundation

Acknowledgements Jenna Hunt Bolette Hartmann Jens Juul Holst Cristoph Ullmer, Roche Innovation Center, Basel, Switzerland