Developmental programming of adult obesity and ...Weight C LP C-HC LP -HC Fat pads (mg) 2251 ± 286 2427 ± 259 3247 ± 538 $$$ 5169 5169 ± 460 $$$ * ... of IUGR + catch up growth

Developmental programming of adult obesity and cardiovascular disease

in rodent by maternal nutrition imbalance

Claude Remacle

Institute of Life SciencesUniversité catholique de Louvain,Louvain-la-Neuve, Belgium

Note: for non-commercial purposes only

Florence BieswalVanesa BolSamira BoujendarHanane CheriffSalouah DahriOlivier DumortierIlham El KhattabiKevin GoosseLuise KalbeSiham MerezakPierre RomanusAnne SnoeckNicolas Theys

University of Louvain

Brigitte Reusens

Thomas BouckenoogheLuise KalbeCéline MorrensPierre Nivoit

Dept of Clinical BiochemistryUniversity of Cambridge, UKSuzan Ozanne

INSERM U457, ParisBernadette Bréant

Rowett Institute, AberdeenW.D. Rees

King’s College, London, UKLucilla Poston

Alarming increase of obesity

Alarming increase of obesity

McAllistair & many others, Crit. Rev. Food Sci. Nutr., 2009

InfectionsEpigeneticsMaternal Age Assortative MatingSleep DebtEndocrine DisruptorsPharmaceutical IatrogenesisAmbient Temperature

food practicereduction in physical activity

Intrauterine & intergenerational Effects

The Big Two :

Ten putativecontributors

Rogers & EURO-BLCS, Int. J. Obes. Relat. Metab. Disord., 2003; Mc. Millen & Robinson, Physiol.Rev., 2005

HighHigh birth weight �� HighHigh BMI in adulthood

SmallSmall birth weight �� HighHigh BMI in adulthood

SmallSmall birth weight �� SmallSmall BMI BUTBUT altered distribution of fat mass

and a lean mass decreased in adulthood

Association between birth weightand adult BMI

Birth weight

AdultBMI

Importance of early catch-up growth ?

?

The Dutch Famine

Nov.1944 – Apr.1945

bread potato

sugar beet

Painter, Thesis, 2006

Born before famine

in mid pregnancy

79.0

91.8

26.7

65 %

79

92.4

26.7

63 %

84.2

95.6

75 %

Effect of prenatal exposure to the Dutch famine on size and obesity at age 50 in men

Ravelli et al., Am J Clin Nutr, 1999

Weight (kg)

Waist (cm)

Body Mass Index (kg/m 2)

BMI > 25 kg/m 2

Exposed to faminein late

pregnancyin early

pregnancy

76.8

91

26.6

64 %

28.1

Intrauterine exposure Unexposed p value

BMImale 24.6 25.2 0.39female 26.9 27.0 0.89

Waist:Hip ratiomale 0.86 0.87 0.66female 0.79 0.78 0.49

Subscapular:Tricepsmale 1.26 1.41 0.50female 1.01 0.88 0.17

Stanner & Yudkin, Twin Res, 2001

Fatness parameters in subjects exposed in uteroto starvation during the Leningrad Siege

5 monthscatch-up growth

28 monthsno catch-up growth

Yajnik, Proc Nutr Soc. 2004

Mean level of central obesity at 8 years of ageby tertiles of birth weight and weight at 8 y

Catch-up growth

Birth weight Weight at 8 y

Experimental groups (1)

20%

20% 20% 16%

16%

Gestation Lactation

C

C-Caf

Birth Week 3

Adult

20%

8%

8% 8%

R

R-Caf

8% 16%

16%

16%

16% + Caf

16%

16 % Caf

Week 7

Bieswal & al., Obesity, 2006

Body weight gain

0

100

200

300

400

500

600

700

800

900

0 4 8 12 16 20 24 28 32 36 40 44 48

C

C-Caf

R

R-Caf

Age (weeks)

Wei

ght g

ain

(gr)

Control

Controlcafeteria


20%

20%

20%

20%

Gestation Lactation

C

LC

Birth Weaning

Adult males (6 months)

16%

16%

8%LP 16%20%

8 Pups

4 Pups

4 Pups

20% 20% 16% + Cafeteria

8% 20% 16% + Cafeteria

20% 20% 16% + Cafeteria


C-Caf

LP-Caf

LC-Caf


Catch up growth during lactation

0

10

20

30

40

50

60

70

0 2 4 6 8 10 12 14 16 18 20 22

Days of lactation

Wei

ght (

gr)

CLPLC

birth weight6.9 ± 0.25.7 ± 0.2**5.7 ± 0.1**


Obesity in adult

0

200

400

600

800

Bod

y W

eigh

t (gr

)

c

ba

c

C LP C LPCaf

Experiment 1No Catch-up growth

Experiment 2Catch-up growth

Bod

y W

eigh

t (gr

)0

200

400

600

800

1000

ab b

cd

e

C LP LC C LPCaf

LC

Body weight at 42 weeks


Experiment 1No Catch-up growth

0

4

8

12

16

20

Rel

ativ

e W

AT

wei

ght (

%)

C LP C LPCaf

Experiment 2Catch-up growth

0

4

8

12

16

20

24

C LP LC

Rel

ativ

e W

AT

wei

ght (

%)

*

**

C LP LCCaf

Weight of visceral fat depots

Obesity in adult


Obesity in adult

In rat,

postnatal catch up growth after intrauterine growth

retardation results in higher body mass and obesity, associated with factors favoring cardio-vascular

disease :

Leptin

Plasminogen Activator Inhibitor-1Angiotensinogen

Adiponectin


20% 20%

Gestation Lactation

C

Birth Weaning

Adult males (36 weeks)

16%

8%LP 16%20%

8 Pups

4 Pups

20% 20% 16% + HC

8% 20% 16% + HC


C-Caf

LP-Caf

Bol & al., Amer J Physiol, 2009

In mice ?

20% 20%

Gestation Lactation

C

Birth Weaning

Adult males (36 weeks)

16%

8%LP 16%20%

8 Pups

4 Pups

20% 20% 16% + HC

8% 20% 16% + HC


C-HC

LP-HC


Corn oil: 4-32%

Sucrose: 10%

Water + Emulsifier

HCStandard ChowStandard Chow

1.41 ± 0.03 g

1.25 ± 0.03 g **

11.1 ± 0.8 g

12.9 ± 0.5 g *

Birth Weaning

Body weight (catch up)

C

LP


Body weight (adult)

31.0 ± 0.7 g

31.0 ± 0.8 g

34.5 ± 1.6 g

40.5 ± 1.5 g *

C

LP

-HC9 months

C-HC

LP-HC


Energy intake

Chow HC suppl.

Corn oil: 4-32%

Sucrose: 10%

Water + Emulsifier

-HCStandardStandard ChowChow


C LP LC C-HC LP-HC LC-HC

Dai

ly E

nerg

y in

take

(kJ

)

0

100

200

300

400

500

600

Bieswal et al, Obesity, 2006

Energy intake

Corn oil: 4-32%

Sucrose: 10%

Water + Emulsifier

-HCStandard ChowStandard Chow

Energy intake

The obesity stimulated by early

malnutrition followed by catch up

growth is not obligatorily attributable to

hyperphagia.

Effects of rapid postnatal catch-up growth on the adult phenotype

Jimenez-Chillaron & Patti, Curr Opin Endocrinol Diabetes Obes. 2007

Adiposity : fat pads


Standard ChowStandard Chow HypercaloricHypercaloricWeight CC LPLP CC--HCHC LPLP--HCHCFat pads (mg)Fat pads (mg) 2251 ± 286 2427 ± 259 3247 3247 ±± 538 538 $$$$$$ 5169 5169 ±± 460 460 $$$ * $$$ * ££

% Fat pads% Fat pads 6.8 ± 0.7 7.3 7.3 ±± 0.60.6 8.8 8.8 ±± 1.11.1 12.5 12.5 ±± 0.70.7

(ANOVA 2 : * foetal diet effect; $ hypercaloric die t effect, £ interaction effect)

Adiposity : fat cell size


Pluripotent stem cell

Multipotential mesenchymal

precursor

Determined preadipocyte

Immature multilocular adipocyte

Mature unilocular adipocyte

Pref-1

C/EBP-ββββ

C/EBP-αααα

SREBP-1

C/EBP-δδδδ

PPAR-δδδδ

PPAR-γγγγAdipocyte specific proteins :- lipogenic enzymes (LPL, Malic enzyme, GPDH, …)- secreted proteins (leptin, adipsin, adiponectin, P AI-1)

MITOSESCLONAL

EXPANSIONGROWTHARREST

MATURATION

C

8% 20%

20%20%

LP

Birth Weaning

� Isolation of

perigonadal adipose

tissue

� Digestion,

centrifugation &

filtration

� Seeding of cells (D0)

� Addition of Insulin &

Dexamethasone (D1)

�Differentiation

(D2-D9)

Assessment of

proliferation and

differentiation at

different time points

28 Days

Fat cell precursors

Bol & al., Obesity, 2008

Fat cell precursors

?Adipocyte precursors capacity for proliferation and differentiationin case of growth retardation

NOBieswal & al., J Nutr, 2004

Stromal-vascular fraction of perigenital fat tissue,1 mo-old rat

Fat cell precursors

?Adipocyte precursors capacity for proliferation and differentiationin case of growth retardation followed by catch up growth

YES

Stromal-vascular fraction of perigenital fat tissue,1 mo-old rat


% of differentiation estimated by rating the TG droplets in cells

Enzymatic activities of two markers of differentiation

DAY 4 DAY 9DAY 7

Fat cell precursors


Determination of protein content

during the culture

Determination of DNA content

during the culture

Fat cell precursors

Proliferation


Cell counting at early stages

of culture

Fat cell precursors

Proliferation


Early markers of differentiation

Fat cell precursors

Proliferation vs

Differentiation

rt-RT

-PC

RCyclin D1

SREBP-1c PPAR-γγγγ


Late marker of differentiation

DAY 9

Fat cell precursors

Leptin


Fat cell precursors

A higher proliferative capacity of fat cell

precursors may contribute to the obesity

stimulated by early malnutrition followed

by catch up growth.

Expression of selected genes

in adult adipose tissue

of IUGR + catch up growth mice

by microarray analysis of 89 murine probes

‘Adipochips’


C

LP

C-HC

LP-HC

Genes (#)Genes (#) InteractionInteraction FoetalFoetal HCHC

Cell cycle (1)Cell cycle (1) 11 11

Hormones/growth factors (11)Hormones/growth factors (11) 22 66 99

Transcription factors (3)Transcription factors (3) 11 22

Transporters (4)Transporters (4) 11 22 22

Cell structure (2)Cell structure (2) 11 44

Lipid metabolism (7)Lipid metabolism (7) 11 55 44

Others (2)Others (2) 11 11 22

Statistically significant modification of expression

according to fetal or adult diet (31/89 genes)

‘Adipochips’


‘Adipochips’

maternal hypercaloric

2 3 15

10 3 3

Adipsin AngiotensinogenCaveolin 1Carnitin palmitoyl transferase-2Cystatin CCyclin D3DiOHacetone phosph. ac.transf.Fatty acid binding protein 4Fatty acid synthaseGlut4Glycerol3P-dehydrogenase mitoGlycerol3P-dehydrogenase cytoHaptoglobulinHydroxysteroid dehydrogenaseResistinSecreted acid cystein glycoprot . SREBP-1cTissue Factor


‘Adipochips’

Glucose

Dihydroxyacetone-P

GlucogenesisGlucogenesis

Pyruvate

GlycolysisGlycolysis

GLUT4

SREBP

DHAPTG3PD

GAPT

Triacylglycerol

Glycerol-3-P

Lysophosphatidic acid

PEPCK

Fatty Acyl CoA

BB--OxidationOxidation

TriacylglycerolTriacylglycerol

synthesissynthesis

Malonyl-CoAFAS

CPT2

Fatty Acid

Fatty Acid

Lipoprotein

DOWN


‘Adipochips’

Glucose

Dihydroxyacetone-P

GlucogenesisGlucogenesis

Pyruvate

GlycolysisGlycolysis

GLUT4

SREBP

DHAPTG3PD

GAPT

Triacylglycerol

Glycerol-3-P

Lysophosphatidic acid

PEPCK

Fatty Acyl CoA

BB--OxidationOxidation

TriacylglycerolTriacylglycerol

synthesissynthesis

Malonyl-CoAFAS

CPT2

Fatty Acid

FABP

4

LPL

Cav1

MCAD

Fatty Acid

Lipoprotein

DOWN

UP


Expression profiling

The early imbalanced diet induced obesity

(supplemented with HC diet).Obesity was accompanied by changes

in adipose expression of several enzymes involved in lipid metabolism and changes in hormones/growth

factors

Is early mismatched nutrition (LP/catch-up growth) able to induce obesity-related CVD (hypertension and atherosclerosis)

?

adipocyte

leptinadiponectin

resistin

angiotensinogenPAI-1MCP-1

serum amyloid Aapelintissue factor

C-V system

Meta-analysis :

Difference in systolic blood pressure :- 0.5-2 mm Hgper 1 kg higher birthweight

Programming of hypertension in

Human

Huxley et al., Lancet, 2002

?

Programming of hypertension in Human

Yajnik, Obesity Reviews, 2002

Relation between birthweight, weight at 8 years, and systolic

blood pressure in Indian Children.

Birth weight

Body weight of 8-year-old

12

3123

� Postnatal obesogenic diet increased blood pressure and heart rate in all offspring groups.

� In adult progeny which received unbalanced diet in early life (growth retardation in utero and postnatal catch up growth), blood pressure and heart rate were higher.

� Their circadian rhythm disappeared.� Although LP offspring showed higher plasma

cholesterol than controls, atherosclerosis assessed in aortic roots featured increased plaque area due to obesogenic feeding but not due to early mismatched nutrition.

Obesity & CVD

Documents

Developmental programming of adult obesity and ...Weight C LP C-HC LP -HC Fat pads (mg) 2251 ± 286 2427 ± 259 3247 ± 538 $$$ 5169 5169 ± 460 $$$ * ... of IUGR + catch up growth