SRI Supplement to Reproductive Sciences - Volume 25 Number 1 - March 2018 - 243A

Scientific Abstracts

in placental dysfunction. Further attention should be given to identifying
morphologic parameters that have the potential to aide in prenatal
diagnosis.
*Figure(s) will be available online.

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Predicting Ovarian Hyperstimulation Syndrome in High Responders:
Alterations in Soluble Fms-Like Tyrosine Kinase (sFLT-1) and
Vascular Endothelial Growth Factor (VEGF) Bioavailability.
Mohamad Irani†,2 Evelyn Minis,2 Dimitrios Nasioudis,1 Steven S Witkin,2
Steven Spandorfer*.2 1University of Pennsylvania, Philadelphia, PA,
United States; 2Weill Cornell Medicine, New York, NY, United States.
INTRODUCTION: Ovarian hyperstimulation syndrome (OHSS) is a
serious condition complicating up to 10% of IVF cycles. To delineate a role
for altered angiogenesis in OHSS we compared the baseline serum levels
of VEGF and its soluble receptor sFLT-1, which suppresses angiogenesis
by sequestering free VEGF, in women who developed OHSS and controls.
METHODS: Sera were obtained on the second day of IVF cycle (before
starting stimulation) from 129 high responders (at high risk of OHSS),
72 who later developed OHSS (30 moderate/severe and 42 mild OHSS)
and 57 women who did not develop OHSS. This was a case-control study
where patients were matched for age, BMI, and number of mature oocytes.
sFLT-1 and VEGF concentrations were determined by ELISA and levels
between groups compared by Mann-Whitney test.
RESULTS: There were no differences in age (34.8±8.2 vs. 34.2±4.4
years), body mass index [23.1 (17-37) vs. 22.2 (17.6-36.7) kg/m2],
parity [0 (0-2) vs. 0 (0-3)] or number of mature oocytes [18 (0-32) vs.
16 (0-37)] (P>0.05) between the two groups. Women who developed
OHSS had significantly lower levels of sFLT-1 (233.4 vs. 947.02 pg/
ml, P<0.0001) and a higher VEGF/sFLT-1 ratio (0.1 vs. 0.06, P=0.04)
compared to controls.

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CONCLUSION: A low sFLT-1 level and an elevated VEGF/sFLT-1
ratio suggest that high VEGF bioavailability contributes to induction of
angiogenesis and development of OHSS. Measurement of serum sFLT-1
and VEGF levels and determination of the VEGF/sFLT-1 ratio prior to
an IVF cycle may predict susceptibility to develop OHSS.

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Peripheral Blood Mononuclear Cells (PBMC) Induce Endothelial
Dysfunction in Human Umbilical Vein Endothelial Cells (HUVEC).
Aishwarya Rengarajan†, Ian Bird, Manish Patankar, Derek Boeldt*.
University of Wisconsin Madison, Madison, WI, United States.
INTRODUCTION: Preeclampsia (PE) is a maternal hypertensive
disorder associated with endothelial dysfunction. This leads to decreased
production of the vasodilator nitric oxide due to reduced sustained phase
Ca2+ bursting. PE is also associated with aberrant infiltration of immune
cells, including changes in number and/or proportion of immune cells,
which may be linked with endothelial dysfunction. Immune cells such
as PBMCs can release cytokines like TNFa and IL-6, which are elevated
in PE and can cause decreased Ca2+ bursts in HUVEC. Alternatively,
PBMCs may potentially modify endothelial function due to cell adhesion
or migration with possibly pronounced effects in PE. Ca2+ bursting was
evaluated in HUVECs co-cultured with PBMCs to model endothelial
function in an elevated immune condition, such as in PE. We hypothesize
that PBMCs promote endothelial dysfunction similar to preeclamptic
conditions by inhibiting Ca2+ bursting.
METHODS: PBMCs were isolated from blood of non-pregnant females
using a histopaque gradient centrifugation. HUVECs over 90% confluence
were loaded with Fura-2 for Ca2+ imaging. The imaging protocol (30
minutes each) in sequence was: 100uM ATP, Wash, PBMC addition,
100uM ATP. The number of Ca2+ bursts was compared between the two
ATP treatments for each cell. Fresh or frozen (to reveal any effects of
cryopreservation) PBMCs were used in 10:1 or 25:1 ratio of PBMCs per
HUVEC. To investigate PBMC stimulated Ca2+response, suramin (100uM)
or U73122 (1uM) was applied before PBMC addition.
RESULTS: With the 10:1 ratio, Ca2+ bursting reduced to 66.7% ± 3.4%
of control (p<0.05) with fresh PBMCs and to 70.7% ± 3.5% with frozen
PBMCs (p<0.05). With the 25:1 ratio, Ca2+ bursting reduced to 71.1%
± 3.3% with fresh PBMCs (p<0.05) and to 64.4% ± 3.6% with frozen
PBMCs (p<0.05). There is no significant difference between the fresh
and frozen PBMCs on Ca2+ bursting and between 10:1 and 25:1. From
data on PBMCs from two frozen sources, 10:1 PBMCs reduced Ca2+
bursting to 71.5±2.4% of control (p<0.05) and 25:1 reduced bursting to
73.2±2.6% of control (p<0.05). Further, PBMC addition by itself could
stimulate a Ca2+ response in HUVECs. U73122 blocks this Ca2+response,
whereas suramin does not.
CONCLUSION: The decrease in agonist stimulated Ca2+ bursting in
HUVECs upon PBMC addition is similar to other models of PE endothelial
dysfunction. This Ca2+ bursting decrease indicates that PBMCs can
potentially promote endothelial dysfunction by decreasing endothelial
vasodilator production capacity. This could therefore be a useful model for
studying the immune cell effects on endothelial function in preeclampsia.
U73122 and suramin data suggests that non-purinergic receptor-mediated
Phospholipase-C signaling is involved in the PBMC stimulated Ca2+
response. Further mechanistic studies on PBMC mediated endothelial
effects are warranted.

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Pregnancies of Female Fetuses with Uncomplicated Term Outcomes
Have Higher mRNA Expression of FOXP3 in First Trimester Decidual
Tissue. Jelmer R Prins†, Tom E Kieffer†, Marijke M Faas, Jan Jaap H
Erwich, Sicco A Scherjon, Sanne J Gordijn. University MC Groningen,
Groningen, Netherlands.
INTRODUCTION: Several associations have been found between fetal
gender and complicated pregnancy outcomes. Male fetuses are more
often born preterm, and pregnancies with male fetuses are more often
complicated by preeclampsia and gestational diabetes mellitus. The
pathophysiological pathways responsible for these associations are not

Friday Posters

Placental Expression of Angiogenic and Invasive Genes is Reduced
in TGA Births. Jennifer A Courtney†, Kathryn Owens, Helen N Jones*.
Cincinnati Children's Hospital Medical Center, Cincinnati, OH, United
States.
INTRODUCTION: Transposition of the Great Arteries (TGA) is a
multifactorial CHD occurring in approximately 0.25% of live births.
TGA pregnancies are associated with preeclampsia. Two-stage surgical
intervention is required during the first year to correct TGA, with postsurgical survival over 90% in the US. We have previously shown that
placentas in babies with TGA and other complex CHDs have reduced
vasculature compared to controls.
METHODS: In the current study, RNA was isolated from TGA (n=10) and
control (n=7) placentas and sequenced by Illumina HiSeq (UC Genomics
Core). Gene analysis was performed using TopHat, R and MSigDB.
Pathway analysis was performed using the PANTHER Classification
System and the PANTHER Overrepresentation Test.
RESULTS: Expression of several genes involved in angiogenesis and
heart development (ADGRE1, FGF7, Podoplanin, and Ceruloplasmin)
was reduced in TGA placentas compared to controls. Panther analysis
showed a 28-fold change in enrichment for the angiogenic pathway
(p<0.05). Two of the identified angiogenic genes, Podoplanin and FGF7,
are also involved in cell migration. Podoplanin, a transmembrane mucinlike protein which is thought to promote fetal angiogenesis and has
been shown to increase angiogenesis in vitro, was significantly reduced
in TGA placentas vs controls (p<0.05). Expression of FGF7, a growth
factor involved in cell migration and tissue repair, was also lower in TGA
vs control (p=0.07). There were no overrepresented pathways in genes
identified as upregulated in TGA vs. control.
CONCLUSION: Transcriptome analysis shows that disruption of
angiogenic and cell migration pathways may lead to impaired placental
function and important changes in the placental physiology of TGA
pregnancies. Lowered angiogenesis and cell migration are linked to
preeclampsia, which is seen in a subset of TGA pregnancies. These
results highlight the important relationship between placental and heart
development.

Reproductive Sciences Vol. 25, Supplement 1, March 2018

Table of Contents for the Digital Edition of SRI Supplement to Reproductive Sciences - Volume 25 Number 1 - March 2018