SRI Supplement to Reproductive Sciences - Volume 25 Number 1 - March 2018 - 175A

Scientific Abstracts

T-197
A New Model of Cervical Damage during Pregnancy and Its Effects on
Timing of Delivery. Ioannis Pavlidis†, Heather MacPherson, Rosemary
Leask, Sarah EM Howie*, Jane E Norman*, Sarah J Stock*. University
of Edinburgh, Edinburgh, United Kingdom.
INTRODUCTION: Ascending infection through the cervix is the most
common cause of preterm birth. There is a need for clinically relevant
animal models to study the interplay between cervical damage, ascending
infection and PTB. Nonoxynol-9 (N-9) is a surfactant that damages
the cervical epithelium in non-pregnant mice. This might enable the
development of a new model of cervical damage during pregnancy.
METHODS: Metabolic activity and Permeability assays were used
to determine N-9 effects on endocervical cell viability and monolayer
permeability. To assess effect on cytokine secretion, N-9 pre-treatment
was followed by LPS challenge. A cytokine array determined secreted
cytokines. IL-6 and IL-8 levels were confirmed by ELISA. N-9-based
solutions were intravaginally administered on pregnant mice for 8h. A
new histological scoring system was developed to describe morphological
damage (AB/PAS staining) and neutrophil infiltrations (anti-Ly6G IHC).
To assess the effect of N-9-mediated damage on timing of delivery,
N-9-based solutions were intravaginally administered on pregnant mice
(gestational D17). To assess whether N-9-mediated damage facilitates
vaginal LPS-induced PTB, mice were either treated with N-9+LPS (D17)
or pre-treated with N-9 for 16h before administered LPS (D17).
RESULTS: N-9 reduces cell viability in a time- and dose-dependent
manner(n=4) and increases monolayer permeability(n=4, P<0.05).
No effect was found on LPS-stimulated cytokine secretion(n=4).
H&E staining showed signs of morphological damage and neutrophil
infiltrations in the cervix and vagina of N-9-treated mice(n=3-4mice/
group). All N-9 doses result in morphological damage in both cervix
and vagina(P<0.05). 10% N-9 resulted in neutrophil infiltrations in both
cervix and vagina(P<0.05). N-9 treatment did not result in PTB (n=5-8). In
addition, it did not facilitate vaginal LPS-induced PTB when administered
with (n=5-8) or before LPS (n=10-14).
CONCLUSION: We describe for the first time a mouse model of
cervical damage during pregnancy. We characterised the model using a
new histological scoring system that accurately describes our findings
and assessed its effects on PTB. This mouse model could have broad
applications in studying the role of the cervix as a barrier to ascending
infection.

T-198
Influx of Leukocytes into the Progesterone-Primed Endometrium is
the Primary Trigger for Endometrial IGFBP-1 Production. Karen C
Wheeler†,2 Margaret Bruce,2 Sabita Dhal,2 Ov Slayden,1 Nihar R Nayak*.2
1
Oregon Health Sciences University, Portland, OR, United States; 2Wayne
State University, Detroit, MI, United States.
INTRODUCTION: Progesterone (P) is known to stimulate
decidualization of endometrial cells and insulin-like growth factor binding
protein-1 (IGFBP-1) production. However, we found a strong upregulation
of IGFBP-1 in the premenstrual/menstrual stage endometrium, when
P is approaching its nadir. We hypothesized that IGFBP-1 production
is not regulated by P alone, but by the combination of hormonal and
inflammatory signals produced by infiltrating leukocytes.
METHODS: Ovariectomized macaques were treated sequentially with
subcutaneous implants of estradiol (E) for 14 days and E plus P for 14
days. P was then withdrawn from all animals to induce menstruation.
Endometrial samples were collected during the second cycles from the
various menstrual stages. In another set of animals, P implants were
added back 24 hours after the initial P withdrawal. Early pregnancy
decidual tissues were obtained from macaques on post-implantation
days 16-22. Human endometrial samples were obtained from normally
cycling women at the time of hysterectomy, and decidual samples were
obtained from elective termination of pregnancy. In situ hybridization

175A

(ISH) and immunohistochemistry (IHC) were performed in full-thickness
endometrial sections to evaluate IGFBP-1 mRNA and protein localization,
and IHC for CD45 was performed for identification of endometrial
leukocytes.
RESULTS: In macaques, IGFBP-1 signals were not detectable by ISH
and IHC in the proliferative or secretory phase endometrium. However,
stromal cell production of IGFBP-1 was dramatically increased in both
decidual tissue (when P Is high) and premenstrual/menstrual phase
endometrium (when P is low), both at the mRNA and protein levels.
Identical results were obtained in the human endometrium/decidua. In
both human and macaques, there was a significant correlation between
IGFBP-1 mRNA expression (silver grain count) and the number of CD45positive endometrial/decidual leukocytes. Interestingly, P replacement
within 24 hours of the initial P withdrawal in macaques completely
suppressed menses and reversed the infiltration of leukocytes into the
endometrium. However, P withdrawal and add back failed to suppress
IGFBP-1 expression.
CONCLUSION: Our results suggest that the influx of leukocytes into
the endometrium/decidua is the key trigger for IGFBP-1 production and
induction of endometrial decidualization in P-primed endometrium. We
also conclude that IGFBP-1 induction in stromal cells is an irreversible
process.

T-199
A Comprehensive RNA Sequencing Analysis Reveals Novel
MicroRNA Sequence in Endometrial Receptivity. Kadri Rekker†,1,5
Signe Altmäe,1 Marina Suhorutshenko,1,5 Maire Peters,1,5 Juan F MartinezBlanch,4 Francisco M Codoñer,3 Felipe Vilella,2,3 Carlos Simón,2,3,6
Andres Salumets,1,5 Agne Velthut-Meikas.1 1Competence Centre on
Health Technologies, Tartu, Estonia; 2Igenomix SL, Valencia, Spain;
3
Instituto Universitario IVI/INCLIVA, Valencia, Spain; 4Lifesequencing
SL, Valencia, Spain; 5University of Tartu, Tartu, Estonia; 6Valencia
University, Valencia, Spain.
INTRODUCTION: Endometrium undergoes extensive changes to
prepare for embryo implantation. There are many players contributing
to the regulation of endometrial receptivity and microRNAs (miRNAs)
are thought to have a significant role in this process. However, there is
no consensus about the miRNAs involved in endometrial receptivity and
clearly further investigation is required. Therefore, we aimed to analyse
the complete endometrial and blood miRNome from pre-receptive and
receptive phase samples to find novel miRNA sequences that reflect the
changes during the endometrial transition to the receptive state.
METHODS: A total of 78 endometrial and 78 blood samples collected
from healthy women at pre-receptive (LH+2) and receptive (LH+8) cycle
phase were subjected to Illumina small RNA sequencing. Identification
of novel miRNAs from sequencing reads was performed by miRDeep2
algorithm. Custom TaqMan Small RNA assay was used for the validation
of novel miRNA (chr2_4401) levels from paired LH+2 and LH+8
endometrial samples.
RESULTS: 11 novel miRNAs were determined from endometrial
tissues and 10 novel miRNAs from blood. Most importantly, chr2_4401
(provisional name) was detected from 66.7% of LH+8 (26/39) and only
2.6% of LH+2 (1/39) endometrial samples. chr2_4401 was not found from
blood. Validation by qPCR showed that the level of chr2_4401 miRNA
is almost 40-fold higher in LH+8 compared to LH+2 endometrium (p <
0.0001). No human miRNA with high similarity sequence for the predicted
novel miRNA was found from miRBase database, however, chr2_4401
shares the seed region (2-7 nt) with miR-200 family miRNAs. Target
prediction for chr2_4401 revealed several genes that have been linked to
endometrial receptivity, including MAP kinases.
CONCLUSION: A comprehensive RNA sequencing analysis uncovered
an interesting novel miRNA chr2_4401 that is highly upregulated in the
receptive endometrium. Further studies are ongoing to find out the specific
role of this miRNA in endometrial tissue.

Thursday Posters

blastocysts. We hope to develop a table to show patients the anticipated
number of "biopsiable" embryos based on morphology of embryos on
Day 3.

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

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SRI Supplement to Reproductive Sciences - Volume 25 Number 1 - March 2018 - Cover3
SRI Supplement to Reproductive Sciences - Volume 25 Number 1 - March 2018 - Cover4
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_december2020
https://www.nxtbook.com/nxtbooks/sage/psychologicalscience_demo
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_october2020
https://www.nxtbook.com/nxtbooks/sage/fai_202009
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_august2020
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_june2020
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_april2020
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_february2020
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_december2019
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_october2019
https://www.nxtbook.com/nxtbooks/sage/fai_201909
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_july2019
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_june2019
https://www.nxtbook.com/nxtbooks/sage/canadianpharmacistsjournal_05062019
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_april2019
https://www.nxtbook.com/nxtbooks/sage/sri_supplement_201903
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_february2019
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_december2018
https://www.nxtbook.com/nxtbooks/sage/tec_20180810
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_october2018
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_julyaugust2018
https://www.nxtbook.com/nxtbooks/sage/fai_201807
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_june2018
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_april2018
https://www.nxtbook.com/nxtbooks/sage/sri_supplement_201803
https://www.nxtbook.com/nxtbooks/sage/slas_discovery_201712
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_february2018
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_december2017
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_november2017
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_october2017
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_september2017
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_julyaugust2017
https://www.nxtbook.com/nxtbooks/sage/fai_supplement_201709
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_june2017
https://www.nxtbook.com/nxtbooks/sage/hospitalpharmacy_may2017
https://www.nxtbook.com/nxtbooks/sage/fai_201706
https://www.nxtbook.com/nxtbooks/sage/fai_201607
https://www.nxtbookmedia.com