SRI Supplement to Reproductive Sciences - Volume 25 Number 1 - March 2018 - 183A

Scientific Abstracts

F-001

F-002
Cortisol Causes Drastic Upregulation of MMP-7 in Human Amnion
Fibroblasts. Luyao Wang†,3 Wangsheng Wang†,1 Hao Ying,2 Kang
Sun*.1 1Ren Ji Hospital, Shanghai Jiao Tong University, Shanghai,
China; 2Shanghai First Maternity and Infant Hospital, Tongji University,
Shanghai, China; 3Shanghai First Maternity and Infant Hospital, Tongji
University, Shanghai, China.
INTRODUCTION: Preterm premature rupture of fetal membranes
precedes 30-40 per cent of preterm births and carries significant risks
for the newborns. Activation of matrix metalloproteinases (MMPs) is
the major cause of extracellular matrix (ECM) degradation in membrane
rupture. Cortisol regeneration in the amnion is linked with a number
of parturition-pertinent events including stimulation of prostaglandin
synthesis and ECM remodeling. However, whether cortisol regeneration
has a role in the regulation of MMPs in the amnion is not known.
METHODS: Primary human amnion fibroblasts were isolated and
cultured from the amnion without labor to study the regulation of MMP
by cortisol. Human amnion tissues were collected from deliveries at term
in the presence and absence of labor to study the effect of labor on MMP.

183A

RESULTS: By using cultured amnion fibroblasts, we found that MMP7 was one of the MMPs that were induced by cortisol. Cortisol (0.01,
0.1 and 1 μM) increased MMP-7 mRNA and protein abundance in a
concentration-dependent manner. Furthermore, cortisol is much more
potent than interlukin-1β in the induction of MMP-7 expression in
amnion fibroblasts. Cortisol at 1 μM, a concentration seen in the amnion
in parturition, increased MMP-7 mRNA by 2063 + 176 fold, while
interlukin-1β (1ng/ml) increased MMP-7 mRNA by only 3.31 + 0.33 fold.
Moreover, MMP-7 abundance increased significantly along with cortisol
regeneration in the amnion tissue during labor. Bioinformatic analysis
revealed an AP-1 binding site in MMP-7 gene promoter. Knocking-down
the expression of AP-1 transcription factors c-Fos or c-Jun significantly
attenuated the induction of MMP-7 expression in amnion fibroblasts.
Cortisol activated AP-1 by increasing c-Jun phosphorylation and c-Fos
expression respectively in amnion fibroblasts. MMP-7, also known as
matrilysin, is a secreted zinc- and calcium-dependent endopeptidase
that degrades a broad range of extracellular matrix substrates including
laminin, fibronectin, proteoglycans, elastin as well as type IV collagen,
the scaffold for the assembly of other ECM components. We found that
neutralization of MMP-7 with its antibody blocked the degradation of
type IV collagen by cortisol in amnion fibroblasts.
CONCLUSION: Cortisol regeneration is a crucial cause of MMP-7
upregulation in the amnion in parturition. The drastic upregulation of
MMP-7 by cortisol is mediated at least in part by c-Fos and c-Jun.

F-003
Progesterone Effects on Cervix Ripening and Preterm Birth in a
Murine Model for Progesterone Withdrawal. Steven M Yellon*, Elaine
J Oldford†, Anne C Heuerman†. Loma Linda University, Loma Linda,
CA, United States.
INTRODUCTION: Ovarian progesterone sustains pregnancy in mice,
while the local loss of progesterone (P4) efficacy is hypothesized to
promote cervix remodeling well before labor and birth. Although P4
treatments delay preterm birth (PTB) after ovariectomy (Ovx) in rodents,
births occur after a few days, at term, or not at all. Whether P4 delays
characteristics of cervix ripening and overrides the proposed local P4
withdrawal to block ripening and birth was the focus of the present study.
METHODS: On day 16 post-breeding (pb), mice were Ovx and given
silastic capsules filled with vehicle or P4 (Ovx+P4, 1g/ml sesame oil, s.c.).
Postmortem, blood for serum P4 and cervix were taken from groups of
mice for study of collagen structure (picrosirius stain birefringence), as
well as stroma densities of cell nuclei (hypertrophy) and macrophages
(inflammation) as before (methods in PMID27233754).
RESULTS: P4 treatment sustained serum concentrations >7-fold higher
than in Ovx mice, and comparable to ovary-intact Sham controls. Preterm
delivery occurred within 24h of Ovx (day 17 pb, n=5), but in the Ovx+P4
group, births were delayed to term (day 19 pb) or did not occur by D20 pb
(n=6 each). In the cervix of Sham controls, reduced cell nuclei density and
collagen degradation occurred with the approach of term as in previous
studies. However, neither characteristic was altered by Ovx compared to
the reduced collagen degradation in Ovx+P4 mice. Thus, withdrawal of
systemic P4 or P4 treatment did not affect these remodeling characteristics.
As for F4/80 macrophages, their density was similarly elevated in the
cervix of Sham and Ovx+P4 mice on days 16.5 pb and PP (term) compared
to that in Ovx mice whether before or after PTB. Fetuses from Ovx+P4
mice were stillborn or did not survive in utero the lengthened pregnancy.
CONCLUSION: Findings support the hypothesis that P4 blocks PTB
induced by loss of systemic P4. Thereafter P4 does not affect the structural
transition from a soft to ripening cervix. Birth in Ovx+P4 mice at term
indicates a functional loss of P4 efficacy for parturition to proceed.
However, in other Ovx mice that remained pregnant, P4 treatment
appeared able to sustain a quiescent uterus to term, and possibly beyond.
While the importance of P4 regulation of macrophage residency or
activities in the prepartum cervix remains to be determined, P4 block of
PTB may prove of benefit to some women at risk for advanced cervix
ripening and premature birth that result from macrophage-mediated
inflammation. (Supported in part by NIH HD054931)

Friday Posters

The Unfolded Protien Response-Mediated Uterine Secretome: A Role
for Tocolysis. J Ingles†,2 A Simpson†,1 C Kyathanahalli*,2 S Hassan*,2
J Pancharatnam*,2 J Condon*.2 1University of Detroit Mercy, Detroit,
MI, United States; 2Wayne State University, Detroit, MI, United States.
INTRODUCTION: In this study, we aim to characterize the components
of the uterine secretome activated during pregnancy and the role it plays in
maintaining quiescence. Recently, our laboratory has identified activation
of the unfolded protein response (UPR) in the pregnant myometrium as
being critical for the maintenance of uterine quiescence. Our evidence
demonstrates cells undergoing activation of the UPR transmit a distinct
set of proteins, which in the extracellular compartment capacitate the
transmission and propagation of the UPR derived tocolytic signal in a
paracrine and endocrine manner.
METHODS: Telomerase immortalized human myometrial cells
(hTERTHM) initially underwent stable isotope amino acid labeling in
culture (SILAC). Specifically, hTERTHM cells were passaged seven times
in heavy or light SILAC media. Cells then underwent UPR activation
by exposure to Tunicamycin, 5.0μg/ml, 1hr or vehicle. SILAC labeled
proteins transmitted from the UPR activated myocyte into the media
were analyzed via LC/MS/MS to define the UPR generated secretome.
In a separate experiment the conditioned media was incubated with a
secondary set of naïve hTERTHM cells and hPBMC's which were then
examined for the potential of the UPR mediated secretome to propagate
the UPR mediated tocolytic signal.
RESULTS: LC/MS/MS identified over 60 validated secreted proteins,
which were bone-fide components of the UPR activated uterine myocyte
secretome. UPR activation up-regulates secreted proteins that are largely
associated with adaptation to pregnancy, anti-inflammatory action and
tocolysis. The most upregulated proteins is GRP78/BiP, which has
the extracellular ability to be an active component of the prosurvival
mechanism that allows for the propagation of tocolytic non-apoptotic
caspase-3 activity. Additionally, atrial natriuretic factor, which has been
demonstrated to induce smooth muscle tocolysis in a paracrine manner,
was enriched. Accordingly, UPR action also down-regulates secreted
proteins that are largely associated with proapoptotic signaling and proinflammatory response. Thrombospondin 1, down regulated over 65-fold
acts as a potent activator of apoptotic caspase-3 and has previously been
associated with the onset of parturition. Furthermore, our analysis revealed
that the UPR mediated secretome actively propagates activation of the
UPR in naïve untreated uterine myocytes and human PBMC's.
CONCLUSION: These data confirm that generation and transmission
of the uterine secretome and propagation of the UPR derived tocolytic
signal is largely an UPR regulated mechanism. We propose that potential
endocrine/paracrine transmission and propagation of the myometrial
UPR allows for local uterine myocyte tissue type fidelity and systemic
conditioning of the vasculature and immune response during pregnancy.

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