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Effect of maternal obesity and preconceptional weight loss on foeto-placental growth and offspring health in mice: expression of epigenetic modifiers at the interface
with metabolism
Polina Panchenko, Mélanie Jouin, Sarah Voisin, Sofiane Safi-Stibler, Marie-Christine Lacroix, Luc Jouneau, Karine Badonnel, Nicolas Meunier,
Hélène Jammes, Claudine Junien, et al.
To cite this version:
Polina Panchenko, Mélanie Jouin, Sarah Voisin, Sofiane Safi-Stibler, Marie-Christine Lacroix, et al..
Effect of maternal obesity and preconceptional weight loss on foeto-placental growth and offspring health in mice: expression of epigenetic modifiers at the interface with metabolism. Epigenetic Inher- itance Symposium 2019, University of Zurich [Zurich] (UZH). CHE., Aug 2019, Zurich, Switzerland.
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1-UMR INRA, ENVA «Developmental Biology and Reproduction»
2-UR INRA «NeuroBiology of Olfaction»
Université Paris-Saclay
Centre INRA de Jouy-en-Josas, Domaine de Vilvert 78350 Jouy-en-Josas
EFFECT OF MATERNAL OBESITY AND PRECONCEPTIONAL WEIGHT LOSS ON FOETO-PLACENTAL GROWTH AND OFFSPRING HEALTH IN MICE:
expression of epigenetic modifiers at the interface with metabolism
Introduction: intergenerational transmission of obesity ... what about preconception weight loss?
Results: Obesity induced foetal growth restriction, transcriptional changes and worsen diet-induced obesity in adulthood. Maternal weight loss is beneficial with some possible adverse outcomes.
Conclusions, discussion and future prospects:
P.E. Panchenko¹, M. Jouin¹, S. Voisin¹, S. Safi-Stibler¹, M.C. Lacroix², L. Jouneau¹, K. Badonnel², N. Meunier², H. Jammes¹, C. Junien¹, C. Baly² and A. Gabory¹
Illustrations macrovector - fr.freepik.com
The epigenome is reprogrammed during gametogenesis and development, untill final differentiation of tissue and continue to be modified with ageing. This plasticity
makes the epigenome a good candidate to participate in DOHaD.
Alteration of the epigenome early during ontogenesis, could be a mechanism of
“memorization” of the in utero environment, contributing to particular gene expression patterns and thus to adult phenotype establishment.
We recorded E18.5 mouse development and measured the mRNA expression of epigenetic modifiers and metabolic genes in foetal liver and placenta. Offspring born to ‘control’, ‘obese’ or ‘weight-loss after diet-induced obesity’
mothers were put on a control or high-fat diet, and we tracked their metabolic parameters and olfactory behaviour.
Gametogenesis Development Post-natal period
Epigenetic reprogramming maternal
obesity/weight loss maternal
metabolism/diet maternal
metabolism/diet
Epigenetic marks inheritence of the early environmental exposure
Adulthood and ageing Health
diseaseand
individual’s environmentself
Maternal obesity is associated to a wide range of fertility troubles, obstetrical and gestational metabolic complications [1]. The babies are also at risk for stillbirth, growth phenotype (small or large birthweight), congenital malformations [2-4]. Moreover, according to the ‘developmental origins of health and disease’ concept, maternal obesity predisposes the offspring to adult onset non-communicable diseases [5].
In human cohorts, it may have positive or negative effects on fetal growth [6-11]. Interestingly, none of these studies followed up the metabolic development of these children and adolescents and additional studies are therefore needed to reveal the long-term health profiles of offspring born to obese mothers who lost weight prior to conception [12-13].
In animal, preconceptional weight loss has been assessed in rodents, sheep and non human primates (macaque). In two rat models, a nutritional intervention in obese dams was beneficial to the offspring, even if all parameters were not normalized [14-15]. In sheep, a strict nutritional intervention before mating had long-term benefits on offspring weight gain but deleterious effects on offspring stress response and glucose metabolism [16-17].
A preconceptional weight loss is widely recommended to obese women. However, its long-term outcomes on the offspring have been poorly assessed.
Before Healthy diet After
F0 F1
5w 14w 23w sacrifice
birth weaning PND21 gestation
6m
2m 3m 4m 5m
Weight Loss (WL)n=26
WL-CD WL-HFD
HFD CD
(n=16 o, 20 o ) (n=19 o, 17 o ) Obese
(OB)n=72
OB-CD OB-HFD HFD
(n=10 o, 9 o ) (n=9 o, 10 o ) Control
(CTRL)
n=40
CTRL-CD CTRL-HFD CD
(n=20 o, 17 o ) (n=20 o, 15 o )
C57Bl/6J
4 Offspring long-term outcomes
We tracked metabolic and olfactory behavioural trajectories of offspring born to CTRL, OBor WL mothers. After weaning, the offspring were either put on a CD or a HFD.
The offspring’s own diet explained most of the variability in metabolic and olfactory phenotypes. After only few weeks of HFD, the offspring developed obesity, metabolic alterations and olfactory impairments, independently of maternal context.
However, maternal obesity had a sex-specific conditioning effect:
Male offspring born to obese mother gained even more weight under HFD than their counterparts born to lean mothers.
Preconceptional WL normalized the offspring metabolic phenotypes but had an unexpected programming effect on olfactory performance:
a reduction in olfactory sensitivity, along with a lack of fasting-induced, olfactory-based motivation.
-13%
1 Maternal phenotype
2 Offspring foetal phenotype
Body weight (g)
5 10 15 20
25 30 35
20 15
abab ab
ab
abc
abc
abc
Obesity (+23%) Dyslipidemia
Glucose intolerance Obesity (+23%) Dyslipidemia
Glucose intolerance
Obesity (+23%) Dyslipidemia
Glucose intolerance
Overweight (+5%)
Metabolic normalisation
After 4 months of HFD, OB mothers presented the characteristics of the metabolic syndrome. Diet change induced weight loss, normalisation of lipidic and glucidic metabolims, although WL mother kept a slight 5% overweight.
statistics: p<0.05 for a OB vs CTRL, b WL vs CTRL, c WL vs OB.
Maternal obesity is associated with fetal growth restriction (-13%) and increased proportion of small for gestational age foetuses (odd ratio =3.2).
Maternal preconceptional weight loss lead to a complete normalisation of the foetal growth phénotype.
In our model, it is unclear whether the decrease in olfactory sensitivity in WL males and the resistance of WL-CD to fasting stem from:
- the preconceptional caloric depletion associated with the transition to CD in mothers
- to a maternal stress linked to the transition per se, as stress may lead to intergenerational phenotypes [22-23].
Further studies are needed to discriminate the effect of energy depletion from the effect of stress on the maternal effects reported herein.
Proportion (%)
SGA (<10 percentile) Normal weight
0 20 40 60 80
100 OR = 3.2
CTRL OB WL
E18.5 foetal weight
Fetal weight (g)
a ns c
0.4 0.6 0.8 1.0 1.2 1.4
CTRL OB WL
statistics: p<0.05 for a OB vs CTRL, b WL vs CTRL, c WL vs OB, ns not significant.
Proportion of Small for Gestational Age foetuses
3 Foetal gene expression
The expression of 60 epigenetic machinery genes and 32 metabolic genes was measured in the fetal liver, placental labyrinth, and junctional zone by RT-qPCR using TaqMan Low Density Assay.
Epigenetic machinery (60 genes):
DNA Methylation (13):
DNMTs
Tet methylcytosine dioxygenases
methylated DNA binding proteins Histone Methylation (18):
Lysine demethylases KDMs
Lysine methyltransferases KMTs
Arginine methyltransferases PRMTs Histone Acetylation (29):
Histone deacetylases HDACs
Lysine acetyltransferases HAT
Bromodomaine proteins Brd
Candidate for metabolism/development (32 genes):
Glucidic and lipidic metabolism:
Pparα, Pparγ, Pparδ, Pepck, Pgc-1α, C/Ebp-α, C/Ebp-β, Lpl, Gck, Rev-erbα, Nocturnin, Oxtr, GcGr, Irs-1, Insr
Appetite regulation:
Leptin, Lepr, Pomc, Npy, Bdnf
Glucocorticoïds regulation and response:
11βHsd-1, 11βHsd-2, Gr Serotonine pathway:
Tph1, Slc6a4, Maoa, 5-HT-r2a, 5-HT-r2c Feto-placental development:
Gcm1, Gcgr, Igf2, Igf2r, Slc16a10, Irs-1, Insr
23 genes were affected by maternal weight trajectories in at least one of three tissues. The fetal liver and placental labyrinth were more responsive to maternal obesity than junctional zone.
One third (18/60) of the epigenetic machinery genes were differentially expressed between at least two maternal groups.
Genes involved in the histone acetylation pathway were particularly altered (13/18).
In OB group, while most Hdacs were downregulated, Kats and Brd2 were upregulated. In WL group, the expression of only a subset of these genes was normalized.
Labyrinth
Junctional zone
Fetal liver
Kat3b Gck
Hdac6 Kat2a Kat6b Kat13d
Prmt1
Prmt7
Hdac3 Hdac10 Sirt4 Mecp2
Mbd5
Kmt1d Kat1 Kat13b
11Hsdβ1
Irs1 Tph1
Kat3a Brd2 Hdac2 Lepr
Brd2
Hdac2 Kat3a
0.0 0.5 1.0 1.5
2.0 a
CTRL OB WL
Relative expression
0 1 2 3
4 b
CTRL OB WL
Relative expression
0.0 0.2 0.4 0.6
0.8 a
CTRL OB WL
0.0 0.2 0.4 0.6
0.8 b
CTRL OB WL 0.0
0.5 1.0 1.5
2.0 b
CTRL OB WL
Labyrinth Fetal liver
0.0 0.2 0.4
0.6 b
a
CTRL OB WL
statistics: p<0.05 for a OB vs CTRL, b WL vs CTRL, c WL vs OB.
Body weight (g)
Age (weeks) 10
20 30 40 50
3 5 10 15 20 25
ac
ac
acacacacaca
c acacac acac
*
Body weight (g)
Age (weeks) 0
10 20 30 40
3 5 10 15 20 25
ac ac
*
CTRL CD HFDOB CD HFDWL CD HFD
25 30 35 40 45 55 50
a
* * c *
CTRL CD HFDOB CD HFDWL CD HFD
25 30 35
20 45 40
* * *
CTRL-CD OB-CD WL-CD
CTRL-HFD OB-HFD WL-HFD
Statistics
a - OB vs CTRL b - WL
c - WL vs OBvs CTRL * - HFD vs CD
Electroolfactogram (EOG) recordings demonstrated a decreased sensitivity of the olfactory epithelium of male offspring born to WL mothers, whaterver their post-weaning diet.
statistics: p<0.05 for a OB vs CTRL, b WL vs CTRL, c WL vs OB, $ dose
response. Heptaldehyde concentration (vol/vol)
Amplitude (mV)
0 5 10 15 25 20
0.00001 0.0001 0.001 0.01 0.1
$
b
-2 0 2 4
-2-10123
Dim 1 (24.24%)
Dim 2 (17.75%)
WL
OB CTRL
-2 0 2 4
-2-1012
Dim 1 (37.51%)
Dim 2 (17.55%)
OB CTRL WL
Female Male
CD
HFD
-2 0 2
-2024
Dim 1 (32.16%)
Dim 2 (20.36%)
WLOB CTRL
-3 -2 -1 0 1 2 3
-10123
Dim 1 (28.12%)
Dim 2 (14.39%)
CTRL
OB
WL
Multiple factor analysis (MFA) confirmed that post weaning diet has a major effect on F1 phenotype and HFD-fed males are additionally influenced by maternal group.
MFA analysed a set of 18 variables regrouped in 3 variable categories:
"Biochemistry" grouped the last measure of fasting glycaemia, insulinaemia, cholesterolemia and leptinemia, and glycemic AUC in OGTT.
"Biometry" grouped the organ to body weight ratio for BAT, Pr WAT, Pg-WAT, Sc-WAT, Total-WAT, Liver, Heart and Kidney, and week-24.5 body weight.
"Behaviour" grouped the week-23 relative caloric intake, day-1 hidden-cookie test retrieving time, HFD filled hole preference ratio and EOG at dose 0.001.
Controles Obeses Weight Loss
Mothers
Offspring
Small for Gestational Age Dysregulation of some
epigenetic modifiers gene expression
(mainly histone acetylation)
Increased diet-induced obestity in males
Normalisation of the foetal growth
Incomplete normalisation of gene expression
Normalisation of body weight Unexpected olfactory phenotype (motivation, EOG)
Ac
Deciphering epigenetics mechanisms in DOHaD.
Memory of weight loss or stress?
Most of the research projects studying the effect of maternal obesity focuses on DNA methylation. However our results, as well as other publications, point out that the histone acetylation pathway can be a key component [18-21].
This highlights the importance of investigating the mechanisms of regulation of histone marks in response to environmental insults. The link between histone modifiers, histone acetylation levels, and placental and hepatic function should be established.
Bibliography
1-Lee and Koren, J Obstet Gynaecol. 2010; 2-Rajasingam et al., Am J Obstetrics and Gynecology 2009; 3-Gaudet et al., BioMed Res Int 2014; 4-Acosta et al., 2015; 5-Hanson and Gluckman, Physiol Rev 2014; 6-Villamor and Cnattingius, Lancet 2006; 7-Kjaer and Nilas, Acta Obstet. Gynecol. Scand. 2013; 8-Smith et al., J. Clin. Endocrinol. Metab. 2009; 9-Jain et al., Am. J. Obstet. Gynecol. 2013; 10-McBain et al., Eur. J. Obstet. Gynecol. Reprod. Biol. 2016; 11-Diouf et al., Eur. J. Epidemiol. 2011; 12-Forsum et al., Food Nutr. Res. 2013; 13-Matusiak et al., J. Obes. 2014; 14-Nathanielsz et al., Nutr. Rev. 2013; 15-Dennison et al., Front. Endocrinol. 2017; 16-Zhang et al., Exp. Diabetes Res.
2011; 17-Nicholas et al., PLoS ONE 2013; 18-Aagaard-Tillery etal., J Mol Endocrinol. 2008; 19-Strakovsky etal., J Physiol. 2011; 20-Suter et al., FASEB J. 2012; 21-Suter etal., Am J Obstet Gynecol. 2014; 22-Taouk and Schulkin, J Dev Orig Health Dis 2016; 23-Zhang et al., Endocrinology 2013
References
Expression of epigenetic machinery genes is sensitive to maternal obesity and weight loss in relation to fetal growth in mice.
Panchenko PE, Voisin S, Jouin M, Jouneau L, Prézelin A, Lecoutre S, Breton C, Jammes H, Junien C, Gabory A.
Clin Epigenetics. 2016
Effect of Maternal Obesity and Preconceptional Weight Loss on Male and Female Offspring Metabolism and Olfactory Performance in Mice.
Panchenko PE, Lacroix MC, Jouin M, Voisin S, Badonnel K, Lemaire M, Meunier N, Safi-Stibler S, Persuy MA, Jouneau L, Durieux D, Lecoutre S, Jammes H, Rousseau-Ralliard D, Breton C, Junien C, Baly C, Gabory A.
Nutrients. 2019
Fundings
This study was supported by the Institut National de la Recherche Agronomique (INRA), the Fondation Cœur et Artères and the Institut Benjamin Delessert. PEP, SV and SSS were supported by the Ministère de la Recherche.
