Fetal survivin Fetal survivin concentrations in normal concentrations in normal

and complicated and complicated pregnancies at termpregnancies at term

Ariadne Malamitsi-PuchnerAriadne Malamitsi-Puchner 11 , Maria Boutsikou, Maria Boutsikou 11 , Sophia , Sophia LiosiLiosi 11 , Dimitr ios Gourgiotis, Dimitr ios Gourgiotis 22 , Venetia-Maria Vraila, Venetia-Maria Vraila 22 , , Dimitr ios HassiakosDimitr ios Hassiakos 11 , Stavroula Baka, Stavroula Baka 11 , Despina D. , Despina D.

BrianaBriana 11

1.1. Neonatal Division, 2nd Department of Obstetrics and Neonatal Division, 2nd Department of Obstetrics and Gynecology, Athens University Medical School, Athens, GreeceGynecology, Athens University Medical School, Athens, Greece2.Research Laboratories, 2nd Department of Pediatrics, Athens 2.Research Laboratories, 2nd Department of Pediatrics, Athens

University Medical School, Athens, GreeceUniversity Medical School, Athens, Greece

ApoptosisApoptosis

• Programmed cell deathProgrammed cell death• regulates cell proliferation regulates cell proliferation • contributes to tissue and organ contributes to tissue and organ

homeostasis homeostasis • important for the development of normal important for the development of normal

human placenta human placenta

Disturbances in feto-placental Disturbances in feto-placental apoptosis seem to be associated apoptosis seem to be associated with abnormal pregnancy outcomewith abnormal pregnancy outcome

• fetal macrosomia, due to maternal fetal macrosomia, due to maternal diabetes mellitus (DM) diabetes mellitus (DM)

• intrauterine growth restriction (IUGR) intrauterine growth restriction (IUGR)

IUGR/ Fetal macrosomiaIUGR/ Fetal macrosomia

• IUGRIUGR is correlated withis correlated with --increased incidence of apoptosis in IUGR-affected fetal increased incidence of apoptosis in IUGR-affected fetal

membranesmembranes -abnormal -abnormal expression of proteins involved in apoptosis expression of proteins involved in apoptosis

and cell turnover and cell turnover

• Fetal macrosomiaFetal macrosomia is correlated withis correlated with -decreased apoptosis in placental cells-decreased apoptosis in placental cells

SurvivinSurvivin

• 16.5 kD protein16.5 kD protein • member of the inhibitors of apoptosis (IAP) member of the inhibitors of apoptosis (IAP)

familyfamily • prominently expressed in fetal tissues and over-prominently expressed in fetal tissues and over-

expressed in cancer cells expressed in cancer cells • important for normal fetal development important for normal fetal development • plays critical roles in placental cell survival and plays critical roles in placental cell survival and

cytotrophoblast cell differentiationcytotrophoblast cell differentiation

HYPOTHESIS OF THE STUDYHYPOTHESIS OF THE STUDY

• Umbilical cord blood Umbilical cord blood concentrations of survivin in concentrations of survivin in IUGR and large-for-gestational-age (LGA-due to IUGR and large-for-gestational-age (LGA-due to gestational DM) cases may differ from gestational DM) cases may differ from respective concentrations in appropriate-for-respective concentrations in appropriate-for-gestational-age (AGA) controls, since the former gestational-age (AGA) controls, since the former are associated with excessive and reduced are associated with excessive and reduced feto-placental apoptosis, respectively. feto-placental apoptosis, respectively.

AIM OF THE STUDYAIM OF THE STUDY

• Investigate cord blood survivin concentrations in IUGR, LGA and AGA pregnancies at birth

• Correlate determined concentrations with gestational age, gender and mode of delivery..

SUBJECTS OF THE STUDYSUBJECTS OF THE STUDY

• 160 healthy, singleton full-term 160 healthy, singleton full-term pregnanciespregnancies

- - 101 AGA 101 AGA (placental weight: (placental weight: 480-621480-621 g)g)

- - 48 asymmetric IUGR 48 asymmetric IUGR ( placental weight ( placental weight 230-420230-420 g)g)

- - 1111 LGA LGA ((placental weight:placental weight: 650-810 650-810 g)g)

• Apgar scores:Apgar scores:>>8 in 8 in 11stst and and 55 thth minuteminute

Gestation Related Optimal Gestation Related Optimal Weight (GROW)Weight (GROW) computer-computer-

generated programmegenerated programme

www.gestation.net

Custom baseDaysBirthweight (g)Gestation: WeeksBooking weight (kg)Maternal height (cm)SexEthnic groupParity at booking56Customised centile =

CAUSES OF INTRAUTERINE CAUSES OF INTRAUTERINE GROWTH RESTRICTIONGROWTH RESTRICTION

• Preeclampsia (Preeclampsia ( n=19 )n=19 ) • Hypertensive disease of pregnancy Hypertensive disease of pregnancy (n=16(n=16 ))• Various diseases : Various diseases : severe type I DMsevere type I DM

(( n=2) , iron deficiency anemia (n=4), n=2) , iron deficiency anemia (n=4), hypothyroidismhypothyroidism (n=4)(n=4)

• Maternal smoking ( n=10 )Maternal smoking ( n=10 )

DEMOGRAPHIC DATA OF THE DEMOGRAPHIC DATA OF THE STUDY POPULATIONSTUDY POPULATION

IUGR LGA AGAIUGR LGA AGA• Gestational age (weeks) Gestational age (weeks) 3838 .4.4±1±1 .4 * .4 * 3838 .. 4±04±0 .9* .9* 3939 .0.0±±1.0 *1.0 *

• BW (g) BW (g) 256256 66±34±34 3 * 3 * 3781±1733781±173 * * 319319 77±29±29 6*6*

• BW centile BW centile 55.. 33±333±3 .8 * .8 * 9494 .. 1 ±31 ±3 .1 * .1 * 3939 .. 3±233±23 .4* .4*

• Gender (male/female) Gender (male/female) 23/25 9/2 60/4123/25 9/2 60/41

• Mode of delivery (VD/ECS)** Mode of delivery (VD/ECS)** 24/24 6/5 77/2424/24 6/5 77/24

• Parity (1Parity (1stst /other) /other) 3131 // 17 17 7/4 70/317/4 70/31

* * values are mean ± SD**VD: vaginal delivery/ ECS: elective cesarian section **VD: vaginal delivery/ ECS: elective cesarian section

MethodsMethods Blood collected from:Blood collected from:• Doubly-clamped umbilical cords (mDoubly-clamped umbilical cords (m ixed ixed

arteriovenous bloodarteriovenous blood ) – reflecting fetal ) – reflecting fetal statestate

• Determination of plasma survivinDetermination of plasma survivin concentrations by enzyme immunoassayconcentrations by enzyme immunoassay ((Human Total Survivin EIAHuman Total Survivin EIA , , Assay Assay Designs, Designs, 5777 Hines Drive, Ann Arbor, 5777 Hines Drive, Ann Arbor, MI 48108, U.S.AMI 48108, U.S.A ))

• Statistical analysis (non-parametric Statistical analysis (non-parametric tests)tests)

Fig. 1.Fig. 1. Cord blood survivin concentrations Cord blood survivin concentrations in AGA, IUGR and LGA pregnanciesin AGA, IUGR and LGA pregnancies ..

LGA IUGRAGA

Su

rviv

in c

once

ntr

atio

ns

(p

g/m

l)

400

300

200

100

0

ResultsResults

• NoNo statistically significant differences in statistically significant differences in cord blood survivin concentrations cord blood survivin concentrations between IUGR, LGA and AGA groups.between IUGR, LGA and AGA groups.

ResultsResults

• In the three groupsIn the three groups• The effect of group (IUGR, LGA, AGA) The effect of group (IUGR, LGA, AGA) (Fig.1)(Fig.1), ,

birthweight, cbirthweight, customized customized centile, gestational age, gender, entile, gestational age, gender, mode of deliverymode of delivery and parity and parity on circulating survivin on circulating survivin

concentrations concentrations was not significantwas not significant • Reference survivin values (median, range): 138.49 Reference survivin values (median, range): 138.49

pg/mL and 71.54 - 349.89 pg/mL, respectivelypg/mL and 71.54 - 349.89 pg/mL, respectively

Conclusions 1Conclusions 1

• Cord blood survivin concentrationsCord blood survivin concentrations in full-term in full-term pregnancies are independent of intrauterine pregnancies are independent of intrauterine growthgrowth

• Cord blood survivin concentrations probably do Cord blood survivin concentrations probably do not reflect the disturbances of feto-placental not reflect the disturbances of feto-placental apoptosis expected in IUGR and fetal apoptosis expected in IUGR and fetal macrosomia, due to gestational DMmacrosomia, due to gestational DM

Conclusions 2Conclusions 2

• PParity, gender and mode of delivery arity, gender and mode of delivery (vaginal or elective cesarean section) do (vaginal or elective cesarean section) do not seem to have any impact on umbilical not seem to have any impact on umbilical cord blood survivin concentrationscord blood survivin concentrations

Conclusions 3Conclusions 3

• Reference values (median) for Reference values (median) for cord blood survivin concentrations cord blood survivin concentrations at term were 138.49 pg/mLat term were 138.49 pg/mL