1. Molecular signalsMolecular signals
at the maternal-fetal interfaceat the maternal-fetal interface
Sylvie Hauguel-de Mouzon, PhD
Metrohealth Medical Center
Case Western Reserve University,
Cleveland, USA
IASO, Boston May 013

2. F P
The maternal-fetal interface
adapted from http://jimkean.wordpress.com
exposome

3. Endocrine changes
Decreased Insulin sensitivity
Maternal concentration of macro-nutrients unchanged
Increased inflammation
Increased Endotoxemia
Gestational exposome of obese women

4. Maternal obesity modifies feto-placental
growth and body composition
Birth wt (g)
0
1000
2000
3000
4000 *
0
Placenta wt (g)
200
400
600
800 *
Fat mass @ birth (g)
0
100
200
300
400
*
lean obese
n = 789 pregnancies at term delivery
Hauguel-de Mouzon , Catalano (MHMC database)

5. Consequences of obesity in pregnancy
AdiposityInsulin
resistance
Prenatal programming of obesity-diabetes

6. Presentation outline
III – placental signals
II – immune signals
I – metabolic-endocrine signals

7. Lean OW & Obese p
Insulin (µU/ml) 6.3 ± 3.7 14.3 ± 5.6 <0.001
Leptin (ng/ml) 13.1 ± 9.1 35.1 ± 20.5 0.001
IL-6 (ng/ml) 1.2 ± 0.4 3.2 ± 3.7 0.05
CRP(mg/dl) 0.4 ± 0.3 0.7 ± 0.4 0.03
TNF-alpha (pg/ml)
Adiponectin (µg/ml)
1.9 ± 1.3
8.4 ± 1.2
1.8 ± 0.8
11.6 ± 1.3
ns
0.001
Endocrine Exposome
in pregnancy with obesity

8. Leptin in pregnancyLeptin in pregnancy
leptinMOTHER
95 %95 % 5 %5 %
FETUS
325 ng/min
Lepercq , Hauguel-de Mouzon et al., JCEM, 2001
leptin

9. Placental Leptin
in diabetic and obese women
Lepercq et al. Diabetes 1998
LeptinmRNA
(foldchangevslean)
0
1
2
Lean
(14)
Obese
(17)
GDM
(11)
*
3
*

10. Leptin increases fatty acid uptake
in cultured placental cells
Palmiticacid(cpm/106
cells)
0
500
1000
1500
2000
2500
12 hours
*
p = 0.0001
control leptin
3000

11. 0
4
8
12
CTL 1 4 8 24
FoldChangevsbasal
0
10
20
30
*
*
*
PLA2G6
0
4
8
4
8
Time with leptin (hours)
PLA2G5
0
10
20
30
PLA2G2
Leptin stimulates
inflammatory pathways
in placental cells
sPhospholipases TNF-alpha
CTL 1 4 8 24
Time with leptin (hours)

12. Activates mitogenesis
Activates angiogenesis
Activates inflammatory pathways (macrophages)
Activates FFA transport
Excess leptin modifies placental function
???? facilitates fat accretion in the fetus

13. Placenta and adipose tissue have
similar cytokine repertoire
Trayhurn, P. et al. J. Nutr. 2006;136:1935S-1939S

14. Immunodetection of adpN proteinadipose
55
no wash wash
37
kDa
blank
placenta
blood
actin
AdpNcopy/pgRNA
0
500
1000
1500
WAT WBC PLA
2000
C
WAT
MBC
placenta
Source of adiponectin at the maternal-fetal interface

15. copy/pgRNA
0
50
100
150
200
mRNA expression
*
Foldchange
µg/ml
10
15
0
5
protein
*
0
DNA methylation
1.0
1.5
2.0
0.5
*
obese
lean
Adiponectin in pregnnacy
SHM, DS Unpublished
adipose tissue plasma

16. Adiponectin receptors
yesplatform+mauthorauthor

17. System A amino acid transporter isoforms in
placental membranes
from control and fADN infused mice
Rosario F J et al. J Physiol 2012
control mice
adpn mice

18. Review: Adiponectin – Powell et al. Placenta 2013
yesplatform+mauthorauthor

19. Presentation outline
III – placental signals
II – immune signals
I – metabolic-endocrine signals

20. CD68+
placental macrophages in obesity
maternal
BMI 22.5 BMI 30.8 BMI 46.7
labeledarea/section(µm2
)
0
5000
10000
15000
CD68 CD14
*
*
ObeseLean
Placenta 2007

21. P. Cani & N. Delzenne 2009
Dietary exposome targets immune pathways
inflammation
inflammation
TLR4
LPS

22. TLR4
2006, Jason K. Kim
Toll-like Receptor 4 inflammatory signals
LPS

23. plasma(EU/ml)
Increased LPS in maternal plasma
Basu, Hauguel-de Mouzon Obesity 2010
ObeseLean
0.0
0.50
1.0
0.25
0.75
p=0.004

24. TLR4 expression human term placenta
TLR4
cytokeratine 7
Dapi

25. Yang, Hauguel-de Mouzon (ongoing studies)
*
Placenta TLR4 mRNA
0
2
4
6
Lean Obese
Foldchangevsbasal

26. TLR4
2006, Jason K.
Kim
Toll-like Receptor 4 inflammatory signals
LPS
FFA

27. Can maternal lipid diet elicit placental signals ?
mono-unsaturated FA
oleate C18:1
saturated FA palmitate C16:0

28. FoldIncreasevscontrol
Fatty acids stimulate TLR4 expression
in human trophoblasts
Yang, Hauguel-de Mouzon (ongoing studies)
0
10
20
30
40
50
CTL BSA LPS OleatePAlmitate

29. Fatty acids stimulate
cytokine expression in human trophoblasts
FoldIncreasevsctl
0
10
20
30
40
50
Yang, Hauguel-de Mouzon unpublished
CTL BSA LPS OA PA
IL-6 TNF-alpha
0
2
4
6
8
10
CTL BSA LPS OA PA

30. Presentation outline
III – placental signals
II – immune signals
I – metabolic-endocrine signals

31. Two sources of circulating cfDNA in Pregnancy
1
2
2
2 + 1

32. Release of cf DNA by Placental shedding
cell-free DNA
cfDNA in maternal blood is 95 % placental (5% fetal)

33. The placenta of obese women is larger,
Inflammed and more apoptotic
>>>
higher release of cf DNA
Hypothesis:

34. pre-gravid BMI
BMI
delivery
Maternal
weight
delivery (kg)
Adipocyte
diameter
delivery (um)
Plasma
leptin
(ng/ml)
Lean (14) 21.8+2.8 27.5+3.0 72+7.5 340+ 52 30+16
Obese (16) 39.2+6.7 45.7+7.3 121+23 501+ 89 67+23
p-value 0.0001 0.0001 0.0001 0.001
0.0001
Study cohort for cf DNA measures

35. cf DNA in systemic circulation
of obese vs lean womenPlasmacfDNA(pg/ml)
0
100
200
300
400
500
lean obese
GAPDH
DYS14
∗
maternal origin

36. cfDNA(pg/ml)
pre-gravid BMI (kg/m2
) pregnancy BMI (kg/m2) gestational wt gain (kg)
0
200
400
600
800 r= 0.57, p=0.002 r= 0.41, p=0.03
0 10 20 30 40
r= 0.49, p=0.006
0 10 20 30 40 50 0 10 20 30 40 50
Circulating cfDNA and maternal adiposity
GAPDH GAPDH GAPDH

38. A: lean B: obese
lean obese
0
20
40
arbitraryunits
C
∗
Increased apoptosis in adipose tissue of obese women

39. cfDNA Increased fat mass in
obese women
is an impetus for
propagating
inflammatory signals
to the FP unit

40. Endocrine changes
Insulin sensitivity decreased
Metabolic inflammation
Molecular signals from obese women
Endotoxemia
Gut microbiota modified
Placental signals
Inflammation
Crosstalks lipid- immune pathways

41. Placental signals
Excess fetal adiposity

42. TThhaannk Yk Yoouu !!

44. 1. Interactions adiponectin, leptin and resistin on glucose metabolism and insulin sensitivity. The normal values of every
hormone are included in the arrows. Negative sign refers that the substance inhibits (−) or increases (+) the effect.

45. Circulating cell-free (naked) DNA
Placental DNA

46. Foldincreasevscontrol
Palmitate C16:0
Haghiac, Hauguel-deMouzon et al. 2012
MCP1 IL1α TNFα IL8 IL6
Fatty–acids stimulate
cytokine release in human adipose cells

47. Lean (n=12) Obese (n=22) p
Early pregnancy BMI 21.2 ± 1.9 32.3 ± 7.7 <0.001
GA (weeks) 8.5 ± 2.1 8.9 ± 1.9 0.57
Glucose (mg/dl) 72.8 ± 8.5 74.4 ± 11.2 0.66
Total cholesterol (mg/dl) 139. ± 37. 151. ± 36. 0.37
HDL-cholesterol (mg/dl) 43. ± 11. 43 ± 8. 0.96
LDL-cholesterol (mg/dl) 86. ± 31.4 97.6 ± 30.3 0.32
VLDL-cholesterol (mg/dl) 12. ± 3.4 13.4 ± 6.1 0.53
Triglyceride (mg/dl) 75 ± 21.3 83. ± 37. 0.53
FFA (meq/L) 0.6 ± 0.3 0.7 ± 0.3 0.44
Maternal exposome in pregnancy with obesity

48. Gut microbiome in obese and lean twins
Nature 2009 Gordon Knight Turnbaugh
lean
obese

49. 10736
Transfer function
Endocrine function
steroids
HCG/HPL
leptin
cytokines
Utilization of placental energy substrates
ATP
Oxidation
Storage
Lipids
Phospholipids
Glycogen

50. cfDNA(pg/ml)
p=0.99 p=0.65
20 25 30 35 40 45 50
0
50
100
150
200
250
20 25 30 35 40 45 50
cfDNA(pg/ml)
0
100
200
300
pre-gravid BMI (kg/m2
) pre-gravid BMI (kg/m2
)
DYS14 SRY
Circulating fetal cfDNA and maternal adiposity