October 17-21 2015 Baltimore Maryland

O-4 Monday, October 19, 2015 12:00 PM
THE SYNCHRONIZATION OF THE LEIOMYOMA
EXTRACELLULAR MATRIX SIGNALING PATHWAYS OF
SURGICAL SPECIMENS IN RESPONSE TO ULIPRISTAL
ACETATE. M. Malik, a J. Cox, b J. Britten, a A. Patel, c L. K. Nieman, d
W. H. Catherino. a a Obstetrics and Gynecology, Uniformed Services University
of the Health Sciences, Bethesda, MD; b Program in Reproductive and
Adult Endocrinology, Eunice Kennedy Shriver National Institute of Child
Health and Human Development, Bethesda, MD; c Henry Jackson Foundation,
Bethesda, MD; d Reproductive Biology and Medicine Branch, Eunice
Kennedy Shriver National Institute of Child Health and Human Development,
Bethesda, MD.
OBJECTIVE: To elucidate the pathways that may be involved in the action
of Ulipristal acetate, a selective progesterone receptor modulator, on leiomyoma
extracellular matrix (ECM) production and breakdown.
DESIGN: Uterine leiomyomas are characterized by increased stiffness as a
result of excessive and disordered ECM which contributes to the total bulk of
the tumor. Ulipristal acetate reduced leiomyoma size in several randomized
placebo-controlled studies. We have previously demonstrated that Ulipristal
acetate reduces the total ECM that forms the bulk of the tumor in over 60% of
patients studied. Clinical tissue was sent for RNASeq analysis to identify
pathways that may be involved in leiomyoma ECM breakdown. In-vitro analysis
was carried out to analyze the effect of the medroxyprogesterone acetate
(MPA) and Ulipristal acetate.
MATERIALS AND METHODS: RNA isolated from clinically collected
fibroid and myometrial tissue from patients treated with Ulipristal acetate
in a placebo-controlled, randomized trial, underwent RNASeq analysis
(Beckman Coulter Genomics Inc). To further evaluate the impact of Ulipristal
acetate, 3-dimensional leiomyoma cultures were treated with MPA, Ulipristal
acetate, and combinations. The expressions of ECM genes as well
as Wnt/b-catenin pathway genes were analyzed. Proteins were analyzed using
Western Blot.
RESULTS: Both TGFb3 and TGFb1 transcripts were reduced in tumors of
women treated with Ulipristal acetate. The reduction was also observed at
protein levels. Multiple components of the TGFb signaling pathway demonstrated
an overall reduction. Reduced transcript and protein expressions were
observed in matrix metalloproteinases (MMPs), such as MMP-9 (2.38-fold)
and MMP-13 (1.97-fold). Decreased WNT5a protein indicated an involvement
of the WNT/b-catenin pathway that was Supported by changes in
expression of Frizzled gene transcripts such as FZD6. In addition, there
was increased expression of aquaporins and other osmotic stress regulators
such as NFAT5, which are known to be regulated by WNT pathway.
Leiomyoma cells in 3D culture, in response to MPA, demonstrated a
3.23+/-0.15-fold increase in b-catenin expression. Ulipristal acetate alone
and in combination with MPA decreased b-catenin expression below that
of untreated leiomyoma cells.
CONCLUSIONS: TGFb signaling pathway in association with both
canonical and non-canonical WNT pathway may participate in Ulipristal
acetate clinical activity by reducing the total amount of ECM proteins produced,
and by decreasing osmotic stress. Ulipristal acetate may also reduce
the progesterone-mediated fibrosis.
Supported by: This research was Supported by Intramural grant from Uniformed
Services University of the Health Sciences, QP85GF13 and
RO85298815. The research was also Supported, in part, by the intramural
research Program in Reproductive and Adult Endocrinology, NIH; NIH
R21 and EMD Serono.
O-5 Monday, October 19, 2015 12:15 PM
ELECTIVE SINGLE EMBRYO TRANSFERS (ESET) AS COMPARED
TO COMPREHENSIVE CHROMOSOME SCREENING
(CCS). A. R. Anderson, D. Taylor, K. C. Chiles, U. Balthazar,
A. S. Browne. Reproductive Medicine Associates of Texas, San Antonio, TX.
OBJECTIVE: To investigate the association with eSET with optimal embryo
development and CCS testing for embryo selection.
DESIGN: A prospective cohort study.
MATERIALS AND METHODS: CCS aneuploidy testing was performed
on 297 embryo transfers. Alternately, 172 eSET blastocyst transfers were
completed based on at least two high quality day 5 embryos available for
transfer where one is transferred and at least one selected for cryopreservation
on day 5. In the CCS subgroup, 5 to 7 cells were biopsied from day 5 and day 6
blastocysts in a HEPES buffered Medium. Biopsied cells were placed in
separate PCR tubes and analyzed for 24 chromosomes at an outside reference
laboratory. All embryos in the CCS subgroup were subjected to vitrification
for a delayed embryo transfer in a subsequent frozen embryo cycle.
RESULTS: There was no significant difference in ongoing pregnancy,
average age, embryos transferred, or miscarriage rates when eSET criterion
or CCS was utilized for embryo selection. In the eSET group there were 172
transfers with 126 (73%) positive pregnancies, 25 (20%) losses, and 101
(59%) ongoing pregnancies. From 297 CCS embryo transfers there were
209 (70%) positive pregnancies, 47 (22%) losses, and 162 (54%) ongoing
pregnancies respectively with no significant difference between these two
subgroups. However, there was a significant (P