Molecular signals Molecular signals at the maternal-fetal interfaceat the maternal-fetal interface

Sylvie Hauguel-de Mouzon, PhDMetrohealth Medical Center

Case Western Reserve University, Cleveland, USA

IASO, Boston May 013

F P

The maternal-fetal interface

adapted from http://jimkean.wordpress.com

exposome

Endocrine changes

Decreased Insulin sensitivity

Maternal concentration of macro-nutrients unchanged

Increased inflammation

Increased Endotoxemia

Gestational exposome of obese women

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)

Consequences of obesity in pregnancy

AdiposityInsulin resistance

Prenatal programming of obesity-diabetes

Presentation outline

III – placental signals

II – immune signals

I – metabolic-endocrine signals

  Lean OW & Obese p

Insulin (U/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 (g/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

Leptin in pregnancyLeptin in pregnancy

leptinMOTHER95 %95 % 5 %5 %

FETUS

325 ng/min

Lepercq , Hauguel-de Mouzon et al., JCEM, 2001

leptin

Placental Leptin in diabetic and obese women

Lepercq et al. Diabetes 1998

Lep

tin

mR

NA

(f

old

ch

ang

e vs

lea

n)

0

1

2

Lean (14)

Obese (17)

GDM (11)

*

3

*

Leptin increases fatty acid uptake in cultured placental cells

Pa

lmit

ic a

cid

(cp

m/1

06 ce

lls)

0

500

1000

1500

2000

2500

12 hours

*

p = 0.0001

control leptin

3000

0

4

8

12

CTL 1 4 8 24Fo

ld C

han

ge

vs

bas

al

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)

Activates mitogenesis

Activates angiogenesis

Activates inflammatory pathways (macrophages)

Activates FFA transport

Excess leptin modifies placental function

???? facilitates fat accretion in the fetus

Placenta and adipose tissue have similar cytokine repertoire

Trayhurn, P. et al. J. Nutr. 2006;136:1935S-1939S

Immunodetection of adpN proteina

dip

os

e

55

no wash wash

37

kDa

bla

nk

placentab

loo

d

actin

Ad

pN

co

py

/p

g R

NA

0

500

1000

1500

WAT WBC PLA

2000

C

WAT

MBC

placenta

Source of adiponectin at the maternal-fetal interface

co

py

/p

g R

NA

0

50

100

150

200

mRNA expression

*

Fo

ld c

ha

ng

e

g/m

l

10

15

0

5

protein

*

0

DNA methylation

1.0

1.5

2.0

0.5

*

obeselean

Adiponectin in pregnnacy

SHM, DS Unpublished

adipose tissue plasma

Adiponectin receptors

yesplatform+mauthorauthor

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

Review: Adiponectin – Powell et al. Placenta 2013

yesplatform+mauthorauthor

Presentation outline

III – placental signals

II – immune signals

I – metabolic-endocrine signals

CD68+ placental macrophages in obesity

maternal BMI 22.5 BMI 30.8 BMI 46.7

lab

eled

are

a/se

ctio

n (m

2 )

0

5000

10000

15000CD68 CD14

*

*

ObeseLean

Placenta 2007

P. Cani & N. Delzenne 2009

Dietary exposome targets immune pathways

inflammation

inflammationT

LR

4

LPS

TL

R4

2006, Jason K. Kim

Toll-like Receptor 4 inflammatory signals

LPS

pla

sma

(EU

/ml)

Increased LPS in maternal plasma

Basu, Hauguel-de Mouzon Obesity 2010

ObeseLean0.0

0.50

1.0

0.25

0.75p=0.004

TLR4 expression human term placenta

TLR4cytokeratine 7Dapi

Yang, Hauguel-de Mouzon (ongoing studies)

*

Placenta TLR4 mRNA

0

2

4

6

Lean Obese

Fo

ld c

han

ge

vs b

asal

TL

R4

2006, Jason K. Kim

Toll-like Receptor 4 inflammatory signals

LPS

FFA

Can maternal lipid diet elicit placental signals ?

mono-unsaturated FAoleate C18:1

saturated FA palmitate C16:0

Fo

ld I

nc

reas

e v

s c

on

tro

l

Fatty acids stimulate TLR4 expression in human trophoblasts

Yang, Hauguel-de Mouzon (ongoing studies)

0

10

20

30

40

50

CTL BSA LPS OleatePAlmitate

Fatty acids stimulate cytokine expression in human trophoblasts

Fo

ld

Incr

ease

vs

ctl

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

Presentation outline

III – placental signals

II – immune signals

I – metabolic-endocrine signals

Two sources of circulating cfDNA in Pregnancy

1

2

2

2 + 1

Release of cf DNA by Placental shedding

cell-free DNA

cfDNA in maternal blood is 95 % placental (5% fetal)

The placenta of obese women is larger, Inflammed and more apoptotic

>>> higher release of cf DNA

Hypothesis:

pre-gravid BMIBMI

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

cf DNA in systemic circulation of obese vs lean women

Pla

sma

cfD

NA

(p

g/m

l)

0

100

200

300

400

500

lean obeseGAPDH

DYS14

maternal origin

cf D

NA

(p

g/m

l)

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

A: lean B: obese

lean obese0

20

40

arb

itra

ry u

nit

s

C

Increased apoptosis in adipose tissue of obese women

cfDNA Increased fat mass in obese womenis an impetus for propagating inflammatory signals to the FP unit

Endocrine changes

Insulin sensitivity decreased

Metabolic inflammation

Molecular signals from obese women

Endotoxemia

Gut microbiota modified

Placental signalsInflammation

Crosstalks lipid- immune pathways

Placental signals

Excess fetal adiposity

TThhaannk Yk Yoou u !!

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.

Circulating cell-free (naked) DNA

Placental DNA

Fo

ld in

crea

se v

s co

ntr

ol

Palmitate C16:0

Haghiac, Hauguel-deMouzon et al. 2012

MCP1 IL1 TNF IL8 IL6

Fatty–acids stimulatecytokine release in human adipose cells

  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 (1)

Gut microbiome in obese and lean twins

Nature 2009 Gordon Knight Turnbaugh

lean

obese

10736

Transfer function

Endocrine functionsteroids

HCG/HPLleptin

cytokines

Utilization of placental energy substrates

ATP

Oxidation

Storage

LipidsPhospholipids

Glycogen

cfD

NA

(p

g/m

l) 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

cfD

NA

(pg

/ml)

0

100

200

300

pre-gravid BMI (kg/m2) pre-gravid BMI (kg/m2)

DYS14 SRY

Circulating fetal cfDNA and maternal adiposity