maternal malnutrition programmes a prediabeticphenotype in ......iugr after uterine artery ligation...

Maternal malnutrition programmes a prediabetic phenotype

in the progeny

Brigitte Reusens

Institut des Sciences de la vie, Université catholique de Louvain,

Louvain-la-Neuve, Belgium

The Power of ProgrammingMunich, 5-8 Mai 2010

Note: for non-commercial purposes only

Florence Florence BieswalBieswalVanesaVanesa BolBolSamira Samira BoujendarBoujendarHananeHanane CherifCherifSalouaSaloua DahriDahriOlivier Olivier DumortierDumortierIlhamIlham El El KhattabiKhattabiKKevinevin GoosseGoosseSihamSiham MerezakMerezakPierre Pierre RomanusRomanusAnne Anne SnoeckSnoeckNicolas TheysNicolas Theys

University of LouvainUniversity of Louvain

Claude Claude RemacleRemacle

Thomas BouckenoogheThomas BouckenoogheLuise Luise KalbeKalbeCCééline line MorrensMorrensPierre Pierre NivoitNivoit

DeptDept of of ClinicalClinical BiochemistryBiochemistryUniversity of Cambridge, UKof Cambridge, UKSuzanSuzan OzanneOzanne

INSERM U457, ParisINSERM U457, ParisBernadette BrBernadette Brééantant

Lawson Institute, London, OntariLawson Institute, London, OntariooDavid HillDavid HillEdith AranyEdith Arany

StenoSteno Institute, Institute, CopenhagenCopenhagen, DK, DKJorn Jorn NerupNerupTomas Tomas SparreSparre

Early Nutrition Programming & HealthOutcomes in Later Life: Obesity & Beyond

Budapest, April 20-21, 2007

BUDAPEST, HUNGARY

Impaired insulin secretion versus sensitivity:

cause and effect?

Brigitte Reusens

The beta cells

or

the insulin-sensitive target tissues

What is first programmed?

Sequence of the key pathologicalfeatures of type 2 diabetes

Leahy JL, 2005

Aquired organ abnormalities

Protein RestrictionCalorie Restriction

Uterine Artery Ligation

Gestational DiabetesHigh Fat / (High saccharide) DietMaternal Obesity

Animal modelsof early food manipulation

Other specific dietary manipulation

Intrauterine Programmingof the

foetal endocrine pancreas?

Glucagon

Insulin

Mass of ββββ cells

Apoptosis or necrosis

Neogenesis from precursors(Ngn3, Pdx1= differentiation)

Proliferation ofdifferentiated ββββ cells

Low Calorie and Low Protein Diets

0

C 20% protein20% protein

20% protein20% protein

20% protein20% protein 8% protein8% protein

8% protein8% protein

F15 F21

LCLLC

LPL

LP

50% cal.

50% cal.

Dumortier et al., 2007

Gestation

ββββ cell mass (mg)

C LCLCL LPLPL

1

0

*

***

**

Beta cell mass and proliferationF 21.5

Image analysis

Dumortier et al., 2007

0

4

8

12

C LCLCL LPLPL

******

Proliferation (%)

BrdU labelling

Cell differentiation

(after the notch-delta lateral

specification)

NGN3NGN3

Differentiation of the endocrine pancreas

Beta 2

Pax 4

?

Brn 4

MafB

Pax 6

? MafA

Acinar cell

Ptf1a

Mist1

Hnf 6

Duct cell

PP δδδδ αααα

Pdx-1

ββββ

Nkx2.2

Nkx6.1

Pdx-1

Endocrine cells

10th -15th day

NGN3NGN3

Differentiation of the endocrine pancreas

Beta 2

Pax 4

?

Brn 4

MafB

Pax 6

? MafA

Acinar cell

Ptf1a

Mist1

Hnf 6

Duct cell

PP δδδδ αααα

Pdx-1

ββββ

Nkx2.2

Nkx6.1

Pdx-1

Endocrine cells

Cell differentiation

(after the notch-delta lateral specification)

10th -15th days

NGN3 / duct area (µm 2 x100)

*

0

0.1

0.2

C LC LP

0

2

4

6

8

C LC LP

**

PDX-1 / duct area (µm 2 x104)

10

Beta cell markers of differentiation(day 15)

Du

mo

rtie

r e

t a

l.,

20

07

NGN3

PDX1

Numerical density (# /104µm2)

0

4

8

12

16

C LCL LC LPL LP

**

0

1

2

3

4

5

C LCL LC LPL LP

*

Volume density (%)Islet vascularizationD

um

ort

ier

et

al.

, 2

00

7

Reduced ββββ cells Mass

Apoptosis or necrosis

Neogenesis Proliferation

Apoptosis or necrosis

Neogenesis Proliferation

Low calorie Low protein

Islet vascularizationGlucocorticoids

Taurine

Islet vascularizationGlucocorticoids

Taurine

1/2

Insulin secretion by fetal islets in vitro

G5.6 G16.7 +Leu +Arg +Tau +Met0

5

10

15

Insu

lin r

elea

se (

%)

Cherif et al, 1998

Insulin secretion by fetal islets in vitro

G5.6 G16.7 +Leu +Arg +Tau +Met0

5

10

15

Insu

lin r

elea

se (

%)

Islet from LP foetuses secrete less insulin in response to secretagogues

Cherif et al, 1998

Foetal islet cell sensitivityto cytotoxic molecules

Apo

ptos

is (

%)

0

5

10

15

20

- + IL1 + SNP (NO)Merezak et al, 2001

Foetal islet cell sensitivityto cytotoxic molecules

Apo

ptos

is (

%)

**

0

5

10

15

20

**

**

- + IL1 + SNP (NO)Merezak et al, 2001

Beta cells from LP foetuses are more susceptible to cytokines

Acquired cell sensitivityto cytotoxic molecules in vitro

Apo

ptos

is(%

)

0

5

- + IL1+ TNF+IFNMerezak et al, 2004

Acquired cell sensitivityto cytotoxic molecules in vitro

Apo

ptos

is(%

)

0

5

**

- + IL1+ TNF+IFNMerezak et al, 2004

Beta cells from LP adult remain more susceptible to cytokines

General Nutritional Molecular Programming ?

• Epigenetic programming • Mitochondrial programming

Nutritional Mitochondrial Programming ?

IUGR afteruterine artery ligation(Islets)

ATP production

Mitochondrial copy number

Simmons & al., 2005

Nutritional Mitochondrial Programming ?

1,299 / 10,343genes

Genes Microarray, AffymetrixGeneChip Rat Expression 230A probe array

C LP

20 % 8 %

Neoformed islets

Significance Analysis of Microarray (SAM)

7 days in vitro in standard culture medium

Fetal pancreata

Reusens et al, 2008

Nutritional Mitochondrial Programming ?

11% of the genes of which the expression was modified

encode for mitochondrial proteinsReusens et al, 2008

Energy derivation by oxidation of organic

Compounds (19)[9.5% of 196]Chi Sq: 31.2

Energy reserve metabolism (3)[4.9% of 61]Chi Sq: 0.80

Main path of carbohydrate (15)

[11.5% of 130]Chi Sq: 33.41

Cellular respiration (12)[29.2% of 41]Chi Sq: 98.63

Glycogen met.(3)[6.5% of 46]Chi Sq:2.01

Glycolysis (6)[10% of 60]Chi Sq: 10.3

Gluconeogenesis (1)[4% of 25]

Chi Sq: 0.09

TCA cycle intermediate metabolism (3)[11.1% of 27]Chi Sq: 6.2

Pentose-phosphate shunt (1)[10.1% of 10]Chi Sq: 1.71

Aerobic respiration (11)[30.5 of 36]

Chi Sq:95.26

Glycogen catabolism (1)[16.6 of 6]Chi Sq: 3.9

Regulation of glycolysis (1)[20% of 5]

Chi Sq: 5.03

Fumarate metabolism (1)[100% of 1]

Chi Sq: 32.65

Citrate metabolism (1)[20% of 5]

Chi Sq: 5.03

Malate metabolism (1)[11.1% of 9]Chi Sq: 2.1

TCA Cycle (10)[31.2% of 32]Chi Sq: 88.96

Low protein diet targeted the Krebs cycleof the beta cells

Reusens et al, 2008

gestation lactation

3 monthsweaning birth

LP

Maternal malnutrition

LC

20%

8%

50%

C

HF

ATP production in islets of LP adult progeny

3.3 mM16.7 mM

Glucose

0

50

100

150

200

250

300

350

C LP

% o

f con

trol

3.3

mM

**

0

50

100

150

200

250

C LP

% o

f con

trol

3.3

mM

Theys et al, 2009