SRI Supplement to Reproductive Sciences - Volume 25 Number 1 - March 2018 - 256A

256A

Reproductive Sciences Vol. 25, Supplement 1, March 2018

Scientific Abstracts

Table 1. Vaginal Delivery Rates and Starting Station in Active Phase

The proteomic analysis was performed by two-dimensional minimaldifference gel electrophoresis and mass spectrometry. Associations
between proteomic changes and biological and molecular functions were
identified with the Database for Annotation, Visualization and Integrated
Discovery (DAVID), v6.7. The expression levels of ten genes, identified
by significant proteomics analysis, were studied by quantitative PCR
(qPCR). One protein was studied by immunohistochemistry of placental
specimens. Altogether 77 SNPs of 10 genes were studied from DNA of
altogether 343 infants born spontaneously at tem (38 wk + 0 d to 41 wk
+ 6 d). Under Bonferroni correction the significance level was p<6.5 x
10-4. Finally, HTR8/SVneo trophoblast cells were transfected with either
pooled CPPED1 siRNA or control siRNA. Transcriptomes of CPPED1
silenced cells and controls were determined by Illumina HiSeq systems
and submitted to bioinformatics analysis.
RESULTS: CPPED1, a phosphatase previously suggested
dephosphorylating AKT1/PKB, was among the ten proteins associated
with spontaneous birth. qRT-PCR from the basal plate revealed that the
mRNA level of CPPED1 was significantly higher in elective caesarean
deliveries than in spontaneous births. Immunohistochemistry showed
CPPED1 in decidual trophoblasts. Analyses of samples from infants
indicated that single-nucleotide polymorphisms rs11643593 and
rs8048866 of CPPED1 were significantly associated with duration of
term pregnancy. Post-transcriptional silencing of CPPED1 in trophoblast
cell line by siRNAs affected gene expression in pathways associated with
inflammation and blood vessel development, and affected AKT1/PKB
signaling pathway.
CONCLUSION: CPPED1 in decidual trophoblasts has a role in the
induction of spontaneous term delivery.

Starting station <0
(unengaged)

Starting station ≥ 0
(engaged)

Protracted
4-6 hours

20/25 (80%)

20/24 (83.3%)

p=0.8

Protracted >6
hours

18/33 (54.6%)

3/16 (18.8%)

p=0.02

Table 2. Vaginal Delivery Rates and Change in Station in Active Phase
With station change Without station change
Protracted 4-6 hours

23/26 (88.5%)

17/23 (73.9%)

p=0.19

Protracted >6 hours

12/29 (41.4%)

9/20 (45%)

p=0.80

S-006
Comprehensive Transcriptomic Profiling of Preterm Villous Tissue
by RNA Sequencing. Heather Brockway†, Louis Muglia*. Cincinnati
Childrens Hospital Medical Center, Cincinnati, OH, United States.
INTRODUCTION: Preterm birth (PTB), birth before 37 weeks gestation,
is a global public health concern and the leading cause of death for children
under 5 years of age. 70% of PTB is idiopathic and spontaneous. While
risk factors include: smoking, stress, infection, and family history, there is
a lack of understanding regarding the underlying molecular mechanisms
involved in PTB. Previous studies have implicated advanced placental
villous maturation in PTB (Morgan,2013;2014). The aim of this project
is to identify transcriptomic signatures from placental villous tissue
associated with PTB at time of delivery and detect molecular signatures
of advanced villous maturation.
METHODS: We identified a selected cohort of idiopathic, spontaneous
PTB (with and without PPROM, excluding any case with chorioamnionitis)
from various biorepositories and obtained frozen cores of placental villous
tissue from preterm births (n=9, 31-36wks, 5 female;4 male) and term
births (n=9, 38-42wks, 5 female;4 male). Total, ribo-depleted RNA was
sequenced utilizing Illumina HiSeq 2000 technology. Expression levels
normalized by RPKM and log transformed. Differential gene expression
and pathway enrichment analyses were conducted using the AltAnalyze
software suite (Emig et al, 2010) and Panther Classification System V12.0
specifically Reactome pathways (Mi et al, 2016).
RESULTS: Our gene level analyses identified 220 upregulated genes and
11 downregulated genes with a 2-fold or greater change in gene expression
between PTB and term birth placental villous samples. Pathway analyses
on the upregulated genes identified 37 pathways with greater than 2-fold
enrichment (Bonferroni adjusted p<0.05). Of these pathways, 5 pathways
represented activation of the complement cascade (13-22-fold enrichment
Bonferroni adjusted p<0.0005). Other pathways include: interferon
signaling, regulation of IGF transport, extracellular matrix organization,
and cell surface interactions (Bonferroni adjusted p<0.0005).
CONCLUSION: These data indicate an upregulation in the placental
villous of a variety of immune responses in the absence of chorioamnionitis,
specifically the activation of the complement pathways. While previous
work examining maternal serum suggested a role for the complement
pathway in PTB, these data indicate that role likely originates in the
placental villous tissue. However, it is unclear if these results are the
result of etiology or the result of pathological outcome.

S-007
CPPED1 Participates in Regulation of Spontaneous Term Birth. Antti
M Haapalainen, Minna K Karjalainen, Mika Rämet, Mikko N Hallman*.
University of Oulu, Oulu, Finland.
INTRODUCTION: Understanding of timing of human parturition
is incomplete. Therefore carried out proteomic and transcriptomic
analyses of full term placentas from uncomplicated pregnancies. We
further performed genome-wide analysis of fetuses representing either
spontaneous births or elective deliveries without labor. The aim was to
identify protein signatures associated with the onset of spontaneous birth.
METHODS: The placenta samples were collected either from
spontaneous births of elective caesarean sections. Both basal and
apical specimens from altogether 31 term placentas were obtained.

S-008
Cervical ECM Modulation in the Face of Inflammation-Mediated
Preterm Birth. Chellen Cheuk-Lun Liu†, Alexandra R Willcockson†,
Shanmugasundaram Nallasamy†, Tulip Nandu, W Lee Kraus*, Mala
Mahendroo*. UT Southwestern Medical Center, Dallas, TX, United States.
INTRODUCTION: Preterm birth (PTB), a delivery that occurs prior
to a full 37 weeks gestation in humans, occurs at a rate of nearly 10% in
the United States and is a leading cause of infant mortality worldwide.
Among the multiple etiologies, infection contributes to a large number
of PTBs. Regardless of timing be it at term or preterm, the cervix must
remodel to become open and flexible to allow for delivery of the fetus.
A better understanding of the transcriptional and structural perturbations
occurring in response to inflammation will allow for the development of
targeted therapies to prevent PTBs.
METHODS: The following investigations were done using mouse
cervices from gestation day 15 untreated, day 18, day 15 + intrauterine
lipopolysaccharide (LPS), and day 15 + sham surgery groups. PolyA
RNA-Seq was performed to compare the transcriptional signature
between term and LPS-mediated preterm ripening. Imaging techniques
were used to visualize extracellular matrix (ECM) structure and immune
cell localization. Second harmonic generation (SHG) identified collagen
morphology, transmission electron microscopy (TEM) identified elastic
fiber ultrastructure while immunofluorescence (IF) and two photon
excitation fluorescence (TPEF) evaluated elastin morphology. Antibodies
for macrophages and neutrophils were used in IF for immune cell
localization.
RESULTS: Gene Ontology analyses of RNA-seq data demonstrate a
unique transcriptional signature in LPS mediated ripening that includes
an upregulation of ECM-targeted protease genes. Imaging techniques
revealed regionalized changes in collagen morphological features and
disruption of elastic fiber ultrastructure, both in the subepithelial stroma of
the tissue upon LPS treatment. IF staining experiments localized immune
cell clusters in the same region of the cervix upon LPS treatment.
CONCLUSION: Collagen and elastic fiber structure in the cervical
subepithelial stroma is distinctly altered with LPS-mediated preterm
birth and correlates with increased concentrations of immune cells in
this unique zone of the cervical stroma. We hypothesize that proteases
secreted by these immune cells are targeting the collagen and elastic fibers
in a region-specific manner, leading to the altered ECM which results in a

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