SRI Supplement to Reproductive Sciences - Volume 25 Number 1 - March 2018 - 95A

Scientific Abstracts

Reproductive Sciences Vol. 25, Supplement 1, March 2018

GO Name

q-value

Negative Regulation of Epidermal Growth Factor Signaling

1.07E-02

Negative Regulation of Protein Dephosphorization

1.60E-02

Thrombin Receptor Signaling

1.92E-02

Cellular Hyperosmotic Response

2.10E-02

Cell Morphogenesis

2.60E-02

Negative Regulation of Necrotic Cell Death

2.80E-02

Glucocorticoid Signaling Pathway

3.20E-02

Regulation of DNA Dependent Transcription

3.50E-02

Protein Heterooligomerization

3.60E-02

Anatomical Structure Formation Involved in Morphogenesis

3.81E-02

Regulation of Sodium Ion Transmembrane Transporter Activity

4.76E-02

Regulation of Coagulation

4.80E-02

Stem Cell Differentiation

4.20E-2

Regulation of Complement Activity

5.28E-2

CONCLUSION: We identified a candidate set of CMP associated proteins
at 10-12 weeks that demonstrate differential expression in pregnancies
that go on to present with PE. Known protein functions indicate biological
plausibility involving a variety of novel processes.

O-114
Paternal Deficiency of Complement Component C1q Leads to
Pregnancy-Specific Hypertension and Impaired Maternal Vascular
Function: A Unique Pregnancy Model. Robert W Powers*, Judith
Brands, Marcia Gallaher. University of Pittsburgh, Pittsburgh, PA,
United States.
INTRODUCTION: Preeclampsia is a significant complication of
pregnancy and a leading cause of maternal and fetal morbidity and
mortality. The underlying cause(s) of preeclampsia are unclear. Several
animal models of preeclampsia have been investigated, however many
are centered on single factors downstream of impaired placentation which
is thought to initiate the syndrome. The paternal C1q deficiency has
been previously reported to cause pregnancy-specific hypertension and
a preeclampsia-like phenotype in female normotensive (low risk) control
C57B6J mice (or other female mice). The focus of this project was to
further investigate this unique paternal mouse model of preeclampsia.
METHODS: Female C57B6J mice were timed mated wither either male
C57B6J or C1q-/- mice. Endothelial-dependent and independent- vascular
function of mesenteric arteries was assessed using an isometric myograph.
Blood pressure was measured by tail cuff and vascular glycocalyx was
assessed by tracer dilution. Data were analyzed by repeated measures
ANOVA or t-test as appropriate with significance at p<0.05.
RESULTS: Despite being normotensive prior to pregnancy and in early
pregnancy (prior to day 10.5dpc), female C57B6J mice bred to C1q
knockout male develop significant pregnancy-specific hypertension
(148±21/106±24 mmHg) compared to C57B6J females mated to male
C57B6J mice (128±17/83±19 mmHg p<0.01). Female C57B6J mice bred
to C1q knockout males also evidenced significantly blunted endothelialdependent relaxation (57±4%) compared to normotensive pregnant

C57B6J females (88±5%, p<0.001). In addition, female mice with
pregnancy-specific hypertension had significantly smaller pups compared
to pregnant normotensive C57B6J mice (0.78±0.07g vs. 0.83±0.11g,
p=0.02). Lastly, female mice bred to C1q males had evidence of an
abnormal vascular glycocalyx, such that their glycocalyx is significantly
different compared to pregnant normotensive control mice and is more
similar to the glycocalyx measured in non-pregnant mice.
CONCLUSION: The paternal C1q-/- mouse model of pregnancy-specific
hypertension exhibits vascular dysfunction during pregnancy in otherwise
normotensive (low risk) control C57B6J mice. This is an intriguing
pregnancy model that warrants additional investigation.
This project supported by the American Heart Association Go Red for
Women 16SFRN27810001.

O-115
Hypoxia-Mediated LIN28B Downregulation Contributes to
Preeclampsia Pathogenesis by Inhibiting Trophoblast Differentiation
and Inducing Inflammation. John Canfield†, Sefa Arlier, Ezinne Mong,
John Lockhart, Jeffrey VanWye, Ozlem Guzeloglu-Kayisli, Frederick
Schatz, Charles J Lockwood, John Tsibris, Umit A Kayisli, Hana TotaryJain*. Morsani College of Medicine, USF Health, Tampa, FL, United
States.
INTRODUCTION: Preeclampsia (PE) is a major pregnancy
complication leading to maternal mortality. In PE, shallow trophoblast
invasion results in incomplete spiral artery transformation causing
inefficient placental perfusion. The resultant hypoxia reduces trophoblast
fusion (syncytialization). Moreover, systemic endothelial dysfunction and
inflammation elevate maternal blood pressure and trigger hypertension in
PE. As an RNA binding protein, LIN28B inhibits let-7 miRNA maturation
and is a primary modulator of growth and metabolism. The role and
regulation of LIN28B in PE is unknown. We hypothesize that PE related
hypoxia downregulates LIN28B expression, which, in turn, impairs
trophoblast differentiation and inflammation.
METHODS: Microarray analysis compared global gene expression
in primary decidual cell (DC) (n=3) vs. cytotrophoblast (CT; n=3) or
syncytiotrophoblast (ST; n=3) cultures. In situ LIN28B levels were
measured in PE (n=13) vs. normal (n=15) gestational aged-matched
placental tissues by immunohistochemistry, immunoblot and RT-PCR.
In vitro LIN28B levels were determined in hypoxia-exposed human
trophoblastic choriocarcinoma cell lines BeWo (n=3) and JEG3 (n=3).
Regulation of trophoblast differentiation and inflammation mediators were
detected in JEG3 cells following shRNA-mediated LIN28B knockdown.
RESULTS: Compared to DCs, CTs and STs express 30- and 14-fold
higher LIN28B mRNA, respectively (p<0.0001). In situ analysis revealed
that LIN28B mRNA and protein levels are significantly reduced in
PE vs. normal placentas (p<0.05) and that LIN28B immunoreactivity
is predominantly detected in CTs and STs. Hypoxia significantly
decreased levels of LIN28B mRNA and protein as well as syncytin-1
(a syncytialization marker) mRNA in JEG3 and BeWo cell cultures
(p<0.05). Moreover, LIN28B knockdown by shRNA in JEG3 cells
suppressed syncytin-1 mRNA expression, whereas integrin β4 (a noninvasive trophoblast marker) and TNFα (a pro-inflammatory cytokine)
mRNA levels were induced.
CONCLUSION: These results indicate for the first time that LIN28B
mRNA and protein levels decrease in syncytiotrophoblasts and
cytotrophoblasts in PE and that reduced LIN28B expression inhibits
markers of trophoblast invasion and syncytialization as well as increases
inflammation, which may play a crucial role in PE pathogenesis.

O-116
Enhanced Angiogenic Potential of the Human Placental Microvascular
Niche. Yingchun Li, Shuhan Ji, Emily J Su*. University of Colorado
Denver, Aurora, CO, United States.
INTRODUCTION: Mature endothelial cells (ECs) have limited
regenerative capacity, whereas endothelial progenitor cells (EPC) are
capable of homing to sites of angiogenesis. Endothelial colony forming
cells (ECFCs) are a type of EPC that participate in angiogenesis and can
also form de novo blood vessels in vivo. However, various limitations

Friday Orals

were validated by physician reviewers for PE <35 weeks. These were
matched to 50 uncomplicated singleton term deliveries. Controls were
matched on gestational age at sampling (+/- 2 weeks). CMPs from these
specimens were isolated via size exclusion chromatography and analyzed
using global proteome profiling based on HRAM mass spectrometry.
After peptides and proteins were identified and quantified and resulting
AUC ratios were used to determine differential expression between cases
and controls. The identified proteins were subjected to protein complex
expansion to identify meaningful pathways/interactions. Biological
relevance was examined using gene ontogeny (GO) terms.
RESULTS: Cases and controls did not differ by mean age (32 vs. 31;
p=0.50), percent non-white (44 vs 54; p=0.38), percent nullip (24 vs. 28;
p=0.79) but did differ on percent chronic hypertension (12 vs. 0; p=0.01)
and percent prior PE (28 vs. 6; p=0.01). Untargeted analysis identified
>600 unique proteins present in both sample sets at 10-12 weeks. With
a FDR of 0.1, 51 proteins exhibited differential expression in cases vs.
controls. Associated biological functions are noted below:

95A

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