Le « programming » au cours développement: entre environnement et génome

Umberto Simeoni

Service de pédiatrie, DMCP

CHUV & FBM/UNIL

Lausanne

5ème Symposium de la Division d’endocrinologie, diabétologie et obésité pédiatriques

13 octobre 2016, CHUV

Lopez et al, Lancet 2006

The growing burden of noncommunicable diseases (cardiovascular disease, obesity, diabetes,…)

Neonatal Mortality

70%: associated with low birth weight

Epigenetic Landscape and Biological Development:

How the environment affects genes regulation

and development

C Waddington

£ 5

.5

-6.5

-7.5

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-9.5

>9

.5Birthweight (pounds)

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rta

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tio Men

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CORONARY HEART DISEASEStandardised mortality ratios in 10141 men & 5585 women

The effect of finite periods of undernutrition at different ages on the composition

and the subsequent development of the rat Widdowson & Mc Cance, Proc. Roy Soc London 1966

The effect of birth weight on the mortality ratio of coronary heart disease in human

Barker et al, Lancet 1989

Developmental programming:

A process by which a stimulus applied during a criticial period of sensitivity

during development exerts not only short term, but lifelong effects

The amplifying effect of the cycle of reproduction:

Worldwide Type 2 Diabetes Mellitus Epidemic

Altered glucose tolerance

in offspring

Transgenerational transmission

Diabetes in pregnancy

Increased prevalence of T2DM

Kidney: Developmental Programming of Hypertension and of Renal Function:

Low maternal protein diet, IUGR & effect of postnatal overfeeding (OF)

Systolic ABP 1 month

0

2

4

6

8

(m

l/m

in

/k

g)

Control IUGR OF IUGR+OF

CCr 12 months (males)

Boubred et al., Am J Physiol Renal Physiol 2007, 2009

Controls IUGR OF IUGR + OF

Controls

IUGR

OF

IUGR + OF

Systolic ABP 4 months

Proteinuria 4 months CCr 4 months

*

Glomerular sclerosis

Hypertension and Renal Damage:

Effects of high protein intakes

IUGR, Maternal Diabetes (Inborn nephron deficit)

↓ Filtration surface area

↑ Single nephron

glomerular filtration rate

(SNGFR)

↑ Arterial

blood pressure

Proteinuria

Glomerulosclerosis

Glomerular hypertension RAS

Glucose tolerance

0

1

2

3

4

5

t0 t30 t120in

su

lin

(n

g/m

l)

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t0 t30 t120

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ia (

g/l)

Glucose tolerance test

Control LP BET

*

* *

*

*p<0.05

Developmental « programming » &

The Developmental Origins of Health and Disease (DOHaD)

Nutrition,

Taste development

Microbiome

Stress

Toxicants

Non

Communicable

Diseases:

Hypertension

Diabesity

Cancer

Broncho-

pulmonary

disease

Infertility

Neuro-psychic

vulnerability,

Senescence…

Epigenetic changes: DNA methylation,

histones deacetylation/ methylation,

phosphorylation, ncRNA

Conception 1000 DAYS 2nd birthday Adulthood

Normal variations in nutrition, stress, environmental toxicants exposure, Periconceptional & perinatal disorders and interventions: Preterm birth, Intra-uterine growth restriction, Maternal diabetes/overweight, C-section, ART

2nd hits

Epigenetics: a long lasting, heritable molecular translation of early genome-environment interactions, in the absence of gene sequence alteration

Non coding RNAs

Royal gelly fed bees

become bigger, live longer and are fertile.

1800 transcripts altered over 2 X

760 up-regulated 1040 down-regulated

Intrauterine growth restricted (IUGR) neonatal rats:

transcriptome analysis

IUGR vs control rats

IUGR kidneys

Epigenetic effectors: IUGR rats

Analysis of specific groups of genes : epigenetic regulators

Buffat et al, PLoS Genet 2011

Apoptosis and

p53 gene methylation

In IUGR rat kidney

Pham et al, Am J Physiol 2003;285:R962

Intergenerational programming

of impaired nephrogenesis

Harrisson & Langley-Evans, Br J Nutr 2009

Peroxysome proliferator activated receptor – a in offspring of rats fed a protein

restricted (PR) diet during gestation, prevented by folate supplementation (F)

Adapted from Lillycrop et al, 2005 & Burdge et al 2008 , Gluckman NEJM 2008

Standard diet

(HP diet)

13,0 g/dL

(NP diet)

8,7 g/dL

D5 D21 D160

G22

J0

IUGR

-> Postnatal HP diet induces

glomerular hyperfiltration, hypertension and long term glomerular sclerosis Boubred et al, Pediatr Res 2016

Kidney: Effect of Postnatal High Protein Diet in IUGR Rat Pups

« Pup-in-the cup model »

*

*

* HP diet

*

High protein intakes in infants

• Metabolic effects:

– Increased circulating insulin & IGF-1 concentrations, decreased

IGF-BP concentrations

– Increased branched chain amino acids concentrations

Ziegler et al 2006; Socha et al 2011

• Renal and cardio-vacular effects:

– Increased renal osmotic load

– Glomerular sclerosis Boubred et al 2007, 2009

Accelerated growth

Long term insulin-resistance, overweight

Long term altered renal function and structure,

hypertension

Weber et al, Am J Clin Nutr 2014

RCT in the first year of age:

• Higher protein formula (2.05

followed by 3.2g/dL, up )

• Lower protein formula (1.25

followed by 1.6g/dL)

• Breast-feeding (>3 months)

Results at 6 years of age: Higher protein formula -> BMI increased by .51 kg/cm2 -> Risk for obesity x 2.43

Long term effects of high protein intakes on BMI: CHOP trial

Fat intakes during infancy

Dietary fat should contribute to 40-60% energy during the first 6 months of life and should decrease gradually to 35% from 6 to 24 months. FAO, Rome 2010

Diet of 8- to 24-month old children in various industrialized countries contains about 16% energy from proteins and 26-35% from fats; in sharp contrast with the composition of breast milk (low-protein (7%) and high-fat (55%) contents). Butte et al, J Am Diet Assoc 2010

Uauy & Dangour, Ann Nutr Metab 2009

Association of nutrition in early life with

body fat and serum leptin at adult age

Rolland-Cachera MF et al, Int J Obesity 2012

A 1% increased energy intake from fat at 2 years is associated, at the

age of 20 years, with:

lower subscapular skinfold thickness (-2.3% SF, -4.41 to – 0.18,

p = 0.03),

lower fat mass (-0.31 kg, - 0.60 to – 0.01, p= 0.04)

lower serum leptin concentration (0.21 ug/l, -0.39 to 0.03, p= 0.02)

Low fat intakes in the first 2 years of life are associated with higher body

fat, particularly at the trunk site and higher leptin concentrations at adult

age.

Most children consumed the equivalent of more than four times their

protein needs, whereas dietary fat intakes were below current

recommendations

Effects of varying proteins concentrations in infant forumlas

on infants weight between 3 and 12 months of age

Patro-Golab et al, J Nutr 2016

Developmental programming

Quelle ampleur d’effet?

≤67.5

-77

>77

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1

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>13.3

Adult weight (kg)

Mean L/A ratio

Weight at age 2y (kg) p< 0.001

Leptin/adiponectin ratio in young, healthy adults

Prevalence of hypertension

Birth

-weight

(kg)

%

Fifths of current weight (kg)

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Barker DJ, Osmond C, Forsen TJ, Kajantie E, Eriksson JG. N Engl.J Med. 2005;353:1802-9

Conséquences à long terme de la prématurité

Indicators of Glucose Regulation in VLBW Young Adults (Hovi et al, NEJM 2007)

Prevalence of cardio-metabolic complications in adults born preterm

Sipola-Leppanen et al, Am J Epidemiol 2015

Altered adiposity in adult phenotype of preterm infants

Preterm vs Term Subjects Mean difference/ratio p

Total adipose tissue 2.21 L (0.3 – 4.1) 0.03

Abdominal adipose tissue 0.51 L (0.1 – 0.9) 0.007

Systolic blood pressure 6.5 mm Hg (2.2 – 10.8) 0.004

Diastolic blood pressure 5.9 mm HG (1.8 -10.1) 0.006

Intrahepatocellular lipid 3.01 (ratio) (1.78 – 5.28) < 0.001

Tibialis intramyocellular lipid 1.31 (ratio) (1.02 – 1.09) 0.04

Modi N et al Pediatr Res 2011

Greater internal adipose tissue and sinificant urinary metabolome

differences in preterm vs term men.

Total body MRI, MRS, urinary metabolomics

Arterial Blood Pressure in

Young Adults Born Preterm

Postnatal growth

-3

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Birth 4M 12M 48M 6Y 21Y

Z s

co

re

Preterm, heightControl, heightPreterm, weightControl, weight

Systolic blood pressure

correlations

r p

Change in Z-score for weight (4-

12 months)

0.426 0.048

Weight gain : 6-21 years

(absolute)

0.419 0.03

Weight gain : 6-21 years

(relative)

0.582 0.001

Change in Z-score for weight (6-

21 years)

0.381 0.05

Change in BMI (6-21 years) 0.489 0.011

Growth Systolic blood pressure

53*

89*

69*

122* 112

46

85

65

0

20

40

60

80

100

120

140

SBP DBP MBP PP

Arte

ria

l p

re

ss

ure

(m

mH

g)

Preterm

Controls

Tauzin et al, JDOHaD 2014

altered in vitro angiogenic capacity

of low-BW infants cord blood

Endothelial Progenitor Cells

Low birth weight and later hypertension Cord blood circulating endothelial progenitor cells (ECFC)

NBW

LBW Low BW Controls

Ligi et al, Blood 2011

Altered in vivo angiogenic

capacity of ECFCs from low birth

weight subjects, decreased vascular

bed density

Developmentally programmed

pathophysiology of hypertension:

Angiogenesis by human cord

blood endothelial progenitor cells (ECFCs) implanted in nude mice

Low Birth Weight Controls

Ligi et al, Blood 2011 Frizeira et al, Blood 2014

Human Cord Blood Endothelial Progenitor Cells Angiogenic Capacity:

Maturation of the pro-angiogenic AMOT gene methylation rate

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70.00

26 28 30 32 34 36 38 40 42

Mét

hy

lati

on

(%

)

Age gestationnel (semaines)

CpG 2

0.00 5.00

10.00 15.00 20.00 25.00 30.00 35.00 40.00

26 28 30 32 34 36 38 40 42 M

éth

yla

tio

n (

%)

Age gestationnel (semaines)

CpG 3

0.00

5.00

10.00

15.00

20.00

25.00

30.00

26 28 30 32 34 36 38 40 42

Mét

hy

lati

on

(%

)

Age gestationnel (semaines)

CpG 4

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5.00

10.00

15.00

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25.00

30.00

26 28 30 32 34 36 38 40 42

Mét

hy

lati

on

(%

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Age gestationnel (semaines)

CpG 5

0.00

5.00

10.00

15.00

20.00

25.00

26 28 30 32 34 36 38 40 42

Mét

hy

lati

on

(%

)

Age gestationnel (semaines)

CpG 6

-1.5092(P=4e-02) -1.3592(P=8.3e-04) -0.9947(P=6.4e-04)

-0.8804(P=3.1e-04) -0.4616(P=1.6e-02)

Gestational age (wks)

Gestational age (wks) Gestational age (wks)

Gestational age (wks) Gestational age (wks)

SIRT1 decrease correlates with accelerated senescence

in Endothelial Colony Forming Cells (ECFCs) from preterm infants

DNA methylation analysis of

the SIRT1 promoter region

Resveratrol restores SIRT1 expression and cell

proliferation in ECFCs

Friseira & Simoncini et al, Blood 2013

Maternal diabetes: cord blood endothelial

progenitors

In vitro hyperglycaemia

Intra-uterine diabetic environment

Ingram et al, Diabetes 2008

Implication of sperm RNAs in transgenerational

inheritance of the effects of early trauma in mice

Gapp et al, Nature Neurosci 2014

Behavioral and metabolic responses in “unpredictable maternal separation

combined with unpredictable maternal stress” (MSUS) males across generations

and in mice derived from RNA-injected oocytes.

Dad is having a much larger role in the whole process,

rather than just delivering his genome and being done with it.

Une prévention précoce ré-orientée vers la période critique de

vulnérabilité: les futur-e-s mères et pères et la première enfance

• Programmes ciblés d’éducation et

information publique, pour la

nutrition et style de vie (activité

physique, sommeil, addictions)

– en période préconceptionnelle

– nutrition et grossesse: • prise de poids gravidique

• supplémentations en micro-nutriments

– dépistage du diabète gestationnel

– allaitement maternel et optimisation

de la nutrition infantile

• Réduction du risque d’exposition

aux toxiques et contaminants

environnementaux (perturbateurs

endocriniens)

• Approches pharmacologiques

(recherche)

• globales: modificateurs de la méthylation

(folates)

• Spécifiques

• Nécessité de biomarqueurs

précoces

« We affirm a life-long approach with

A focus on the nutrition of women

of reproductive age, pregnant women,

nursing mothers and children under five,

with particular attention to the first 1000

days from pregnancy to a child’s second

birthday. »

Human genome

Genetic variants

Human epigenome

Health vs

Non communicable diseases

Periconceptional period

1000 days Life course

Nutrition, lifestyle, stress

Toxicants & environmental

disruptors

Innate Acquired