nd Annual Minisymposium on Reproductive Biology

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					            32nd Annual
Minisymposium on Reproductive Biology
                          April 17, 2012
     Prentice Women’s Hospital Levine Bernick “L South” Auditorium
        250 E. Superior Street Northwestern University Chicago, IL




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                             Sponsored by:!

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                    About the Center for Reproductive Science
The Center for Reproductive Science (CRS) includes faculty whose research is pushing the boundaries of
reproductive science and medicine. The Center includes faculty from across the University who share common
interests in research and training in the reproductive sciences.

CRS has three missions: 1. Enhance and coordinate research in the reproductive sciences at Northwestern,
2. Promote the application of this research toward human welfare and, 3. Train the next generation of
reproductive scientists, clinicians and educators.

CRS includes 45 faculty from 13 departments in the Weinberg College of Arts and Sciences, the McCormick
School of Engineering, Children’s Memorial Research Center and the Feinberg School of Medicine. Research
areas of current faculty interest include: circadian rhythms; clinical fertility and infertility; gonadal
development and function; growth and development; neural control of sexual function; neuroendocrinology;
ovarian function; prostate biology; reproduction and society; and testis function.

CRS administers a multidisciplinary NIH P01 Program Project, Hormonal Signals that Regulate Ovarian
Differentiation (Kelly Mayo, P.I.), while also facilitating a number of other reproductive science related grants
and training programs. For further information about the center, please visit the CRS website at
http://www.research.northwestern.edu/crs The CRS Newsletter entitled “Reproduction Matters” is a
biannual publication also available on the CRS website.


                                  Center-Sponsored Awards
CRS “Go to Meeting” Award. This is a travel award for undergraduates working in the laboratories of CRS
Members to accompany other lab members to professional meetings such as the annual meeting of the
Endocrine Society, Society for the Study of Reproduction, Society for Neuroscience, American Society of
Reproductive Medicine, etc.

Marcia L. Storch Scholarship for Undergraduate Women Dr. Storch practiced gynecology in New York
City for a number of years and died at her home in Maine. She was always interested in the education of young
women, and stipulated that donations in her name be made to the Center for Reproductive Science for the
purpose of introducing undergraduate women to research. This scholarship is awarded to undergraduate
women working in some aspect of ovarian research and provides a sum for supplies in the designated
laboratory.

Go to www.northwestern.edu/research/crs/ for details on how to apply for the CRS “Go to Meeting” and the
“Marcia L. Storch Scholarship for Undergraduate Women” awards. The deadline to apply for both awards is
June 1, 2012.

Constance Campbell Memorial Research Award. The awards given at the Annual Minisymposium on
Reproductive Biology are named after Constance Campbell. Connie, a Ph.D. in psychology from University
                  of Illinois-Chicago, was hired as an assistant professor in the department of Biological
                  Sciences at Northwestern in 1974. Her research was on the relationship of sexual and
                  other behaviors to environmental cues. She was promoted to associate professor in the
                  department of Neurobiology and Physiology and administered the undergraduate honors
                  program before her untimely death in 1981. Connie cared deeply about student research.
                  After her death, her family, friends and colleagues raised money which is now used to
                  fund the Constance Campbell Memorial Research Awards for the Minisymposium on
                  Reproductive Biology.	
  
                                                 Table of Contents

About the Center/Center Sponsored Awards ................................................... Inside Cover
Neena B. Schwartz Lectureship in Reproductive Science ................................................ 3
Lectureship Recipient, Susan J. Fisher PhD ..................................................................... 5
Northwestern Alumna Speaker, Pamela Kreeger PhD ..................................................... 7
Program for Minisymposium ............................................................................................ 9
ABSTRACTS:
  Oral Session .............................................................................................................. 19
     Poster Session ........................................................................................................... 22
List of Registrants ........................................................................................................... 37
List of Presenters............................................................................................................. 40


                                          Minisymposium Overview
                                               Prentice Women’s Hospital
                                          Levine Bernick “L South” Auditorium


          8:30 – 10 AM                    Searle Center for Teaching Excellence Focus Group

         10:00 AM                         Registration / Light Refreshments
         10:30 AM                         Welcome /Announcements
         10:40 AM                         Northwestern Alumna Speaker – Dr. Pamela Kreeger
         11:10 AM                         Oral Session
         12:20 PM                         Lunch
         1:05 PM                          Poster Session
         2:05 PM                          Introduction to Keynote Address
         2:15 PM                          Keynote Address – Dr. Susan J. Fisher
         3:15 PM                          Awards Presentations
         3:25 PM                          Closing Comments
         3:30 PM                          Adjourn

         4:00 PM                          Center for Reproductive Science Faculty Meeting & Dinner



  Cover Photograph courtesy of the Woodruff Lab and So-Youn Kim, PhD, Northwestern University
                     “Mouse postnatal day 5 ovary stained for MSY2 (green),
                         p63 (red), DNA (DAPI, blue) to highlight oocytes”
	
  
     Neena B. Schwartz Lectureship in Reproductive Science


This year is the 32nd anniversary of the Minisymposium on
Reproductive Biology. The idea for the Minisymposium, a
day-long event focused on providing trainees with an early
opportunity to present their research, came primarily from
Dr. Neena B. Schwartz, William Deering Professor Emerita
of Biological Sciences and founder of the Center for
Reproductive Science.

Neena is a native of Baltimore who has spent most of her
research and teaching career in the greater Chicago area.
After obtaining her undergraduate degree from Goucher College, Neena completed her
MS and PhD. degrees in physiology at Northwestern University. Her first faculty position
was at the University of Illinois College of Medicine, where she rose to the rank of
Professor before moving to Northwestern University in 1974 as Chair of the Department
of Biological Sciences. Neena founded the Program in Reproductive Research, now the
Center for Reproductive Science, in 1980, and developed the program into a premier
training site for students and young investigators interested in reproductive
endocrinology.

Neena has had a distinguished and productive research career and has made many
seminal contributions toward understanding the hypothalamic-pituitary-gonadal axis and
its control. Her early studies on the rat estrous cycle established many of the basic tenets
of cyclical changes in gonadotropin secretion upon which our current views of the HPG
axis are based. Much of Neena’s research has focused on the issue of differential
regulation of pituitary FSH and LH secretion. It was this interest that led her and the late
Cornelia Channing to describe a nonsteroidal feedback factor from the ovary involved in
controlling the secondary FSH surge, and Neena’s laboratory went on to make many
important contributions to the study of ovarian inhibin.

To honor Dr. Schwartz, the Center for Reproductive Science named the keynote address
the Neena B. Schwartz Lectureship in Reproductive Science. This year, Susan J. Fisher
PhD, Professor in the Department of Obstetrics, Gynecology and Reproductive Sciences
and Director of the Human Embryonic Stem Cell Program at University of California,
San Francisco, will give the keynote address and receive the 2012 lectureship award.




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             32nd Minisymposium on Reproductive Biology
         Neena B. Schwartz Lectureship in Reproductive Science




                                     Susan J. Fisher, PhD

        Dr. Susan Fisher is a Professor in the Department of Obstetrics, Gynecology and
Reproductive Sciences and Director of the Human Embryonic Stem Cell Program at the University
of California San Francisco (UCSF). Dr. Fisher has over 25 years of experience studying the human
placenta in the context of normal pregnancy and pregnancy complications. She and her research
group developed in vitro models that have allowed them to study the role of proteinases, cell-matrix
interactions, angiogenic/vasculogenic regulators, and immune and signaling molecules in normal
placentation. This information enabled parallel analyses of abnormal placentation in preeclampsia
and preterm birth, and advanced the concept that both disorders are associated with faulty trophoblast
differentiation. Dr. Fisher also has 33 years of experience in biological mass spectrometry (MS).
Her background spans lipids, protein, and carbohydrate analyses. In recent years, her group has used
MS-based approaches to compile the proteomes of several human organs, tissues, and body fluids.
They have also developed iTRAQ approaches that enable the relative comparison of proteomes. In
addition, her group has played a major role in the NCI-funded Clinical Proteomic Technology
Assessment for Cancer Program’s efforts to advance SID-MRM-MS approaches for protein
quantification.
        Dr. Fisher received her Bachelor of Arts from the University of Michigan in 1973 and her
Doctorate of Philosophy degree in Anatomy from the University of Kentucky in 1977. She then
proceeded to a post-doctoral fellowship at the University of Kentucky in the field of mass
spectrometry. She then joined UCSF in 1982 and has been a faculty member at UCSF for the past 30
years with countless leadership positions and honors. In addition to her academic appointments at
her home institution, Dr. Fisher is an active member of the reproductive biology field nationally. She
organized the first Keystone Symposium on Reproductive Biology as well as the National Academies
Workshop on Stem Cell Models for Environmental Health in Washington D.C. Having published
nearly 200 original research articles, Dr. Fisher has been invited to speak at numerous national and
international meetings on her research.



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            32nd Minisymposium on Reproductive Biology
                    Northwestern Alumna Guest




                                 Pamela K. Kreeger, PhD
                                 Assistant Professor
                                 Department of Biomedical Engineering
                                 University of Wisconsin-Madison

                                 Northwestern Graduate Student-Shea Lab


Pamela K. Kreeger received her PhD from Northwestern University in 2005. As a
member of Dr. Lonnie D. Shea’s laboratory, she studied in vitro ovarian development
using alginate matrices to look at how extracellular matrix components and FSH
stimulation can impact follicle development in culture. During her time as a graduate
student at Northwestern, Dr. Kreeger was the recipient of numerous fellowships and
awards including the Metz Scholarship in Reproductive Biology. After graduating from
Northwestern, Dr. Kreeger went on to a postdoctoral position in cancer biology with Dr.
Douglas A. Lauffenburger investigating RAS signaling networks in apoptosis in in vitro
and in vivo systems. Currently, Dr. Kreeger is an Assistant Professor in the Department
of Biomedical Engineering at the University of Wisconsin in Madison. Integrating her
graduate and postdoctoral work, Dr. Kreeger’s research focus in her lab is in
understanding signaling pathways in ovarian cancers as well as the cell-cell interactions
in endometriosis using experimental and computational analyses.




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                              PROGRAM FOR THE
                  32nd ANNUAL
     MINISYMPOSIUM ON REPRODUCTIVE BIOLOGY
                                     April 17, 2012
                               Prentice Women’s Hospital
                          Levine Bernick “L South” Auditorium
                                Northwestern University
                                       Chicago, IL



8:30 – 10:00 AM      Searle Center for Teaching Excellence Focus Group with Center for
                     Reproductive Science Trainees, breakfast provided - By invitation only.



10:00 - 10:30 AM     Registration                       Atrium
                     Continental Breakfast              L South Auditorium
                     Run-through talks                  L South Auditorium
                     Setup Posters                      Atrium

10:30 AM             Opening Remarks – Irene Lee, Minisymposium Student Committee
                     Chairperson
                     CRS Announcements and Activities - Kelly E. Mayo, PhD, Director,
                     Center for Reproductive Science; Professor in the Department of
                     Molecular Biosciences; Associate Dean for Research, Weinberg College
                     of Arts & Sciences

10:40 AM             Introduction of Alumna Speaker – Pam               Monahan,     PhD,
                     Minisymposium Student Committee member
                     Northwestern Alumna Address – “Biological Engineering Approaches
                     for Women’s Health”, Pamela Kreeger, PhD, Assistant Professor of
                     Biomedical Engineering, University of Wisconsin-Madison




11:05 - 12:20 PM     ORAL SESSION

           Session Moderators: Kristen Meldi, Graduate Student in Mayo Lab, Molecular
           Biosciences Department, Weinberg College of Arts and Sciences and Masanori Ono,
           PhD, Postdoc in Bulun Lab, Obstetrics & Gynecology Department, Feinberg School
           of Medicine



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           Abstract #    * Presenting author underlined

11:10 AM      T1        The role of Geminin in germinal stem cells. Kelly Barry1, K.M.
                        Schultz1, T.J. McGarry1; 1Feinberg Cardiovascular Research Institute,
                        Northwestern University, Chicago, IL.

11:30 AM      T2        Smad-mediated TGFβ signaling regulates epithelial cell fate in the
                        developing vagina through expression of Δ Np63. Monica Laronda1,
                        Kenji Unno1, Lindsey M. Butler1, Vanida A. Serna1, Alea A. Mills2,
                        Richard Behringer3, Chuxia Deng4,Satrajit Sinha5, Takeshi Kurita1;
                        1
                          Division of Reproductive Biology Research, Department of Obstetrics
                        and Gynecology, Feinberg School of Medicine, Northwestern University,
                        Chicago, IL; 2Cold Spring Harbor Laboratory, Cold Spring Harbor, NY;
                        3
                          Department of Genetics, University of Texas, M. D. Anderson Cancer
                        Center, Houston, TX; 4Genetics of Development and Disease Branch,
                        10/9N105, National Institute of Diabetes, Digestive and Kidney Diseases,
                        National Institutes of Health, Bethesda, MD; 5Department of
                        Biochemistry, State University of New York at Buffalo, Center for
                        Excellence in Bioinformatics and Life Sciences, Buffalo, NY.

11:50 AM      T3        AKT inhibitor, MK-2206, reduces leiomyoma cell viability and
                        induces autophagy of uterine leiomyoma cells in-vitro and reduces
                        tumor volume in-vivo. Elizabeth Sefton1, Wenan Qiang1, Vanida Serna1,
                        Takeshi Kurita1, Julie Kim1; 1Department of Obstetrics and Gynecology,
                        Northwestern University.	
  	
  



12:20 - 1:05 PM         Lunch Break



1:05 - 2:10 PM          POSTER SESSION

   P1         Follicle environment-associated alteration of oocyte-specific gene expression
              in oocyte and its polar body: potential markers of oocyte competence. Ze-Xu
              Jiao1, Min Xu1 and Teresa K Woodruff 1; 1Department of Obstetrics and
              Gynecology, Feinberg School of Medicine, Northwestern University, Chicago,
              IL.

   P2         Enhanced survival of mouse ovarian follicles cultured in low oxygen tension.
              Yogeshwar Makanji1, Jennifer Pahnke1 and Teresa K Woodruff1; 1Department of
              Obstetrics and Gynecology, Feinberg School of Medicine, Northwestern
              University, Chicago, IL.




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P3    In vitro ovulation and luteinization of mouse, rhesus, and human follicles in a
      three-dimensional culture system. Robin Skory1 and Teresa K. Woodruff1;
      1
        Department of Ob/Gyn, Feinberg School of Medicine, Northwestern University,
      Chicago, IL.

P4    Disruption of the Notch signaling pathway in the ovary results in aberrant
      follicle formation. Dallas Vanorny1, Signe Kilen1, and Kelly Mayo1; 1Department
      of Molecular Biosciences and Center for Reproductive Science, Northwestern
      University, Evanston, IL.

P5    Zinc maintains prophase I arrest in mouse oocytes through regulation of the
      MOS-MAPK pathway. Betty Kong1, M. L. Bernhardt1, A. M. Kim1, T. V.
      O’Halloran2,3, T. K. Woodruff1,3 1Department of Obstetrics and Gynecology,
      Northwestern University, Feinberg School of Medicine, Chicago, IL 60611;
      2
        Department of Chemistry, Northwestern University, Evanston, IL 60208;
      3
        Department of Molecular Biosciences, Northwestern University, Evanston, IL
      60208.

P6    Repeated phase shifts disrupt pregnancy in the mouse. Keith Summa1,2,
      Martha Hotz Vitaterna1, F.W. Turek1,2, 1Center for Sleep and Circadian Biology,
      Department of Neurobiology, Northwestern University, Evanston, IL,
      2
        Northwestern University Feinberg School of Medicine, Chicago, IL.

P7    Development of ovarian cultures to rapidly test chemotherapeutic
      fertotoxicity in vitro and in vivo. So-Youn Kim1, Jessina Tomas1, Vanida Ann
      Serna1, Marilia Cordeiro1,2, Takeshi Kurita1, Teresa K. Woodruff1; 1Department
      of Obstetrics and Gynecology, Feinberg School of Medicine, Northwestern
      University; 2Center of Neuroscience and Cell Biology, Coimbra, Portugal.

P8    In vitro follicle growth (IVFG) as a novel ovarian function assay in
      reproductive toxicology. Yuanming Xu1, Min Xu1, Teresa K. Woodruff1,
      1
        Northwestern University, Feinberg School of Medicine, Department of
      Obstetrics and Gynecology, Chicago, IL 6061.

P9    Large scale isolation of primordial follicles for transplantation within
      biomaterial grafts in a mouse infertility model. Ekaterina Kniazeva¹, A.N.
      Hardy², L.D. Shea¹, ¹Department of Chemical and Biological Engineering,
      ²Department of Surgery, Northwestern University, Chicago, IL.

P10   Exploring the effects of the Smad4 signaling pathway on Pax2 activated cells
      during early mouse urogenital system development. Jerry Rhee1, Kenji Unno2,
      Vanida Ann Serna2, Lindsey Marie Butler2, Philip M. Iannaccone1 and Takeshi
      Kurita2; 1CMRC/Northwestern Department of Pediatrics, Developmental Biology
      Program; 2Center for Genetic Medicine, Northwestern Feinberg School of
      Medicine, Chicago, IL.




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   P11       Down-regulation of miR-29b is essential for the pathogenesis of uterine
             leiomyoma. Wenan Qiang1, Zhaojian Liu2, Vanida Ann Serna1, Yu Liu1, Stacy
             Druschitz1, Jian-jun Wei2, and Takeshi Kurita1, 1Division of Reproductive
             Biology Research, Department of Obstetrics and Gynecology; 2Department of
             Pathology; Northwestern University Feinberg School of Medicine, Chicago,
             Illinois 60611.

   P12       The role of miR-146a in male germline stem cell self-renewal, Jessica Huszar1
             and Christopher J. Payne1, 1Department of Human Molecular Genetics, Children’s
             Memorial Research Center, Chicago, IL.

   P13       Relating economic conditions to vasectomy and vasectomy reversal rates: a
             bi-institutional study. Sherwin Zargaroff1 , Vidit Sharma1, Kunj R Sheth1, Brian
             V. Le1, James H. Dupree1, Marc Goldstein2, Robert E Brannigan1; 1 Northwestern
             University Feinberg School of Medicine, Department of Urology, 2 Weill Cornell
             Medical College of Cornell University, Department of Urology and Institute for
             Reproductive Medicine and The Population Council, Center for Biomedical
             Research, New York, NY.

   P14       Onco-fertility outcomes may differ by cancer diagnosis, Mary Ellen Pavone1, J
             Hirshfeld-Cytron2, A Lawson1, K Smith1, R Kazer1, S Klock1; 1Department of
             Obstetrics and Gynecology, Northwestern University, Chicago, Il; 2Department of
             Obstetrics and Gynecology, University of Illinois, Chicago, IL.

   P15       National incidence of testicular examination in men ages 15-45: predictive
             factors and barriers. KR Sheth1, Vidit Sharma1, S Zargaroff1, B Le1, JM
             Dupree1, RE Brannigan1, 1Department of Urology, Feinberg School of Medicine,
             Chicago, IL.


2:05 - 2:15 PM      Introduction of Keynote Speaker – Sherwin             Zargaroff,    MD,
                    Minisymposium Student Committee member
                    Presentation of the Neena B. Schwartz Lectureship in Reproductive
                    Science – Kelly E. Mayo, PhD, Director, Center for Reproductive
                    Science; Professor in the Department of Molecular Biosciences; Associate
                    Dean for Research, Weinberg College of Arts & Sciences

2:15 - 3:15 PM      Keynote Address – “Preeclampsia: The Trophoblast Twist”, Susan J.
                    Fisher, PhD, Professor in the Department of Obstetrics, Gynecology, and
                    Reproductive Sciences and Director of the Human Embryonic Stem Cell
                    Program at the University of California, San Francisco

3:15 - 3:25 PM      Presentation of Constance Campbell Research Awards – Kelly E. Mayo,
                    PhD, Director, Center for Reproductive Science; Professor in the
                    Department of Molecular Biosciences; Associate Dean for Research,
                    Weinberg College of Arts & Sciences



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3:25 – 3:30 PM     Closing Comments – Irene Lee, Minisymposium Student Committee
                   Chairperson

3:30 PM            Adjourn


            **** Please take down posters immediately after the program ****




4:00 PM            All Center for Reproductive Science Faculty Meeting and Dinner
                   Lowden Hall, Levy Meyer LM117




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                                                                                    Abstract # T1

The role of Geminin in germinal stem cells. Kelly Barry1, K.M. Schultz1, T.J. McGarry1;
1
  Feinberg Cardiovascular Research Institute, Northwestern University, Chicago, IL.

A proper balance between stem cell self-renewal and differentiation is essential to maintain
tissue homeostasis. The mechanisms that maintain this balance are poorly understood despite
their medical and scientific importance. Spermatogenesis offers a unique opportunity to study
stem cell division and differentiation because cells at all stages of development can be
distinguished microscopically. The unstable regulatory protein Geminin is thought to be one of
the factors that determine whether stem cells continue to divide or terminally differentiate.
Geminin regulates the extent of DNA replication and is thought to maintain cells in an
undifferentiated state by inhibiting various transcription factors and chromatin remodeling
proteins. To examine how Geminin might regulate spermatogenesis, we developed a mouse
model in which Geminin is deleted from germ cells. Geminin disruption leads to the complete
loss of germ cells and sterility in male mice. Geminin(-/-) spermatogonia disappear during the
initial wave of mitotic proliferation that occurs in the first week of life. Geminin is not required
for meiosis or for differentiation of spermatids into mature sperm. Geminin(-/-) spermatogonia
exhibit more double-stranded DNA breaks than control cells, consistent with a defect in DNA
replication. They maintain expression of pluripotency genes and do not prematurely express
genes characteristic of more differentiated spermatogonia. In females, Geminin is not required
for oocyte maintenance after the embryonic period of mitotic proliferation of has ceased. We
conclude that Geminin is absolutely required for mitotic proliferation of spermatogonia but does
not regulate their differentiation. Our results suggest that Geminin maintains replication fidelity
during the mitotic phase of spermatogenesis, ensuring the precise duplication of genetic
information for transmission to the next generation. Supported by NIH grant HL092513A.




                                                                                                 19
Abstract # T2

Smad-mediated TGFβ signaling regulates epithelial cell fate in the developing vagina
through expression of ΔNp63. Monica Laronda1, Kenji Unno1, Lindsey M. Butler1, Vanida A.
Serna1, Alea A. Mills2, Richard Behringer3, Chuxia Deng4,Satrajit Sinha5, Takeshi Kurita1;
1
  Division of Reproductive Biology Research, Department of Obstetrics and Gynecology,
Feinberg School of Medicine, Northwestern University, Chicago, IL; 2Cold Spring Harbor
Laboratory, Cold Spring Harbor, NY; 3Department of Genetics, University of Texas, M. D.
Anderson Cancer Center, Houston, TX; 4Genetics of Development and Disease Branch,
10/9N105, National Institute of Diabetes, Digestive and Kidney Diseases, National Institutes of
Health, Bethesda, MD; 5Department of Biochemistry, State University of New York at Buffalo,
Center for Excellence in Bioinformatics and Life Sciences, Buffalo, NY.

Müllerian duct epithelium (MDE) develops into uterine and vaginal epithelia. Paracrine factors
from uterine mesenchyme induce MDE to remain columnar and form glands, while factors from
the vaginal mesenchyme induce MDE to differentiate as stratified squamous epithelium.
Previously, we demonstrated that induction of transformation related protein 63 (p63) is essential
for MDE to become vaginal epithelium. Furthermore, women exposed to diethylstilbestrol
(DES) in utero have a high prevalence of vaginal adenosis and adenocarcoma, which is the
abnormal development of glandular tissue in the vagina. In this study, we specifically inactivated
the ΔN isoform of p63 in MDE. Vaginal epithelium without the ΔN isoform remained columnar
and did not differentiate into squamous cells, indicating that ΔN is required for vaginal epithelial
differentiation. Microarray analyses were performed to identify mesenchymal factors that
regulate expression of ΔNp63. These analyses identified TGFβ ligands, namely Activin A
(Inhba) and bone morphogenetic protein 4 (Bmp4), as factors that are highly expressed in the
vaginal mesenchyme when compared to uterine mesenchyme. In addition, SMAD4 bound
directly upstream of the ΔNp63 transcription start site in vaginal but not uterine epithelium from
neonatal mice. Furthermore, the vaginal epithelium of Smad4 conditional knockout mice was
negative for ΔNp63 and remained columnar. Our array and immunohistochemistry analyses
demonstrated that RUNX1, a known partner of Smad-mediated transcription, was also highly
expressed in vaginal but not uterine epithelium of neonatal mice. The vaginal epithelium of
Runx1 conditional knockout mice contained dramatically fewer P63-positive cells, and therefore
fewer squamous cells. Thus, we concluded that the epithelial cell fate decision in the vagina is
controlled by activation of ΔNp63 by a SMAD/RUNX complex, which is induced by
mesenchymal Activin A and BMP4. This research is supported by the NIH/NCI R01 CA154358.




20
                                                                                   Abstract # T3

AKT inhibitor, MK-2206, reduces leiomyoma cell viability and induces autophagy of
uterine leiomyoma cells in-vitro and reduces tumor volume in-vivo. Elizabeth Sefton1, Wenan
Qiang1, Vanida Serna1, Takeshi Kurita1, Julie Kim1; 1Department of Obstetrics and Gynecology,
Northwestern University.

Uterine leiomyomas, benign tumors of the myometrium, are the number one indication for
hysterectomies in the United States. Rates of surgery are high due to a lack of effective therapy.
Uterine leiomyomas show increased phosphorylated AKT (p-AKT) compared to normal
myometrium indicating that AKT signaling is increased in the tumor and may contribute to
tumor viability. For this reason, we hypothesized that AKT inhibition would reduce leiomyoma
cell viability. The AKT inhibitor MK-2206 was used to test our hypothesis. MK-2206 (MK)
effectively reduced AKT and AKT substrate, PRAS40, phosphorylation over 24 hours. In
addition, mTOR substrate p70S6K phosphorylation was reduced with MK. Although 10uM MK
over 48 hours reduced leiomyoma cell viability, cell proliferation was unaffected. Despite that
MK reduced viability, little apoptosis was induced as indicated by low cleaved caspase 3
induction and lack of annexin V staining coupled with DAPI positivity. Additionally, MK
induced autophagy in leiomyoma cells as determined by LC3 II induction and punctate LC3
staining. Together these data indicate that apoptosis is not the major contributor to MK reduced
cell viability and that autophagy may have a role in leiomyoma cell death. To determine if MK
is effective in-vivo, primary human leiomyoma cells embedded in collagen were grown under the
kidney capsule of immunocompromised mice and mice were treated weekly with 360 mg/kg MK
for 3 weeks. MK reduced leiomyoma tumor volume in 2 of 3 patients tested compared to vehicle
treated mice. P-AKT was also reduced in lung tissue of MK treated mice compared to the control
group indicating that MK was effective at the dose and treatment schedule. We have
demonstrated that AKT inhibition using MK reduces leiomyoma cell viability, induces
autophagy, and reduces leiomyoma xenograft volume in-vivo. We are actively determining the
function of MK induced autophagy in relation to leiomyoma cell survival and whether mTOR
regulates autophagy in response to MK. Supported by NIH P01HD057877.




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Abstract # P1

Follicle environment-associated alteration of oocyte-specific gene expression in oocyte and
its polar body: potential markers of oocyte competence. Ze-Xu Jiao1, Min Xu1 and Teresa K
Woodruff 1; 1Department of Obstetrics and Gynecology, Feinberg School of Medicine,
Northwestern University, Chicago, IL.

Background: Non-invasive tests of mature egg quality and embryo potential are extremely
limited but urgently needed. Oocyte-specific genes play important roles in oogenesis,
fertilization and at the time of implantation. Our previous data showed that detection and
abundance of oocyte-derived mRNAs in polar body may provide insight into oocyte competence.
We investigate the dynamic expression profiles of oocyte-specific gene in different follicle
environment. At the same, we quantified the mRNA in individual PBs and their sibling oocytes.
The effect of follicle environment on mRNA abundance between PB and oocytes were also
analyzed and compared.
Methods: Secondary mouse follicles were grown in vitro using different concentrations (0.25%
vs. 1.5%) of alignate in a three-dimensional culture system. Oocytes were isolated at the end of
culture and MII eggs matured. The first polar body was removed from all oocytes followed by
reverse transcription and sibling oocytes were similarly prepared. Real-time PCR was performed
to detect and quantify relative gene expression levels. Six oocyte-specific genes (GDF9,
BMP15, Nlrp5, ZP3, H1foo and TCL1) were detected in single oocyte and its sibling polar body.
Results: As expected, we can detect and quantify all of 6 candidate oocyte-specific mRNA
transcripts in individual PBs. All of these 6 candidate genes were differentially expressed
between 0.25% and 1.5% ALG oocyte (P < 0.05). In all cases, the abundance of gene transcripts
for each gene was lower in PBs cultured in 1.5% ALG compared with PBs from 0.25% ALG (P
< 0.05).
Conclusion(s): There is a significant difference in the transcriptional level of oocyte-specific
genes when oocytes are harvested from follicles cultured in different concentrations of alginate.
These data suggest a direct link between the local follicle environment and terminal oocyte
competence. The ability to detect and compare differences in gene expression from individual
PB maybe a potential marker of oocyte competence.
Funding: This project was supported by Grant Number P01 HD021921 from NICHD/NIH.




22
                                                                                   Abstract # P2

Enhanced survival of mouse ovarian follicles cultured in low oxygen tension. Yogeshwar
Makanji1, Jennifer Pahnke1 and Teresa K Woodruff1; 1Department of Obstetrics and
Gynecology, Feinberg School of Medicine, Northwestern University, Chicago, IL.

Folliculogenesis is a complex process controlled by various factors. Unraveling this process
requires a better comprehension of the physiology of follicle growth initiation, interplay of
growth factors, basic metabolic needs of early growing follicles and the physical environment of
the follicle. A proposed activation signal for early follicles is oxygen. A three-dimensional
alginate system for follicle culture has been developed and represents a novel method of
replicating the in vivo environment of the growing follicle. In this study, late primary follicles
were cultured in low oxygen tension to determine the effect of O2 on growth and survival of
mouse ovarian follicles. Follicles (85- 130 µm) were isolated from female CD1 mice and
encapsulated in 0.25% alginate. The follicles enclosed within an alginate bead were cultured in
200 µl of growth media at 2.5, 5 or 20% O2, 5% CO2, 37°C for 8 days. At the end of culture the
size of the follicle was measured using Image J software. The culture media was assayed for
mouse VEGF and estradiol in ELISAs. Follicles that had a starting size of 100 µm had a 80%
survival rate when cultured in 2.5 or 5% O2 compared to 20% survival when cultured in 20% O2.
In addition, growth was improved at lower O2 tension, follicles cultured in 2.5% O2 with a
starting size of 108 ± 10 µm grew to 180 ± 33 µm (67% growth) in 8 days, a similar growth rate
was observed for follicles grown at 5% O2. In comparison, follicles cultured in 20% O2 with a
starting size of 106 ± 6 µm grew to 135 ± 13 µm (28% growth). Interestingly, the survival of
follicles was directly correlated to VEGF levels, follicles cultured in 2.5% O2 had 128 pg/ml, 5%
O2 had 59 pg/ml and 20% O2 had undetectable levels. In addition, estradiol levels were
correlated to follicle size/stage. In conclusion, low oxygen tension (2.5 and 5% O2) improves
survival of late primary mouse follicles and these follicles secrete more VEGF compared to ones
cultured in high O2 tension (20% O2). Supported by NIH funding to the U54 Center for
Reproductive Research, Northwestern University (HD04187) and a NHMRC Overseas
Biomedical Training Fellowship (GNT1016460) to Y.M.




                                                                                               23
Abstract # P3

In vitro ovulation and luteinization of mouse, rhesus, and human follicles in a three-
dimensional culture system. Robin Skory1 and Teresa K. Woodruff1; 1Department of Ob/Gyn,
Feinberg School of Medicine, Northwestern University, Chicago, IL.

Ovulation is a dynamic event thought to require several ovarian microenvironments. As the
capstone of the reproductive cycle, ovulation leads to release of a mature egg and somatic cell
luteinization. The present study sought to identify periovulatory mechanisms specific to the
follicle using the alginate system, which maintains follicular architecture in 3D. To assess
whether follicles ruptured in vitro, murine follicles were cultured in alginate and hCG was added
when diameters reached 285µm. In addition, corresponding oocytes were scored to assess
meiotic competence. Sixteen hours post-hCG, 89% of follicles ruptured (n=38). Of the follicles
that ruptured, 91% released meiotically competent (MII) oocytes. To assess luteinization of the
somatic cell compartment, hCG was added and culture was continued in alginate. Post-hCG,
somatic cells demonstrated characteristics of structural and functional luteinization. Namely,
estradiol synthesis decreased 10-fold and progesterone synthesis increased 80-fold. In addition,
rodent follicles luteinized in vitro showed decreasing levels of inhibin A and B, mimicking in
vivo patterns. To examine luteinization of higher mammalian species, follicles from rhesus and
human were similarly cultured in alginate. In primate follicles, cells luteinized and increased
secretion of progesterone post-hCG administration. Importantly, inhibin A expression increased,
which accurately phenocopies the species difference between rodent and primate. These results
show for the first time that follicular rupture is a follicle-autonomous process, occurring
independent of the reticuloendothelial system or ovarian surface epithelium. In contrast to 2D
cultures, follicles cultured in the 3D alginate system accurately mimicked in vivo hormone
patterns. In the future this system may be used to examine periovulatory hormonal regulation and
to screen novel compounds for contraception. Supported by NIH/NICHD U54HD041857 and
NIH/NIA F30AG040916.




24
                                                                                  Abstract # P4

Disruption of the Notch signaling pathway in the ovary results in aberrant follicle
formation. Dallas Vanorny1, Signe Kilen1, and Kelly Mayo1; 1Department of Molecular
Biosciences and Center for Reproductive Science, Northwestern University, Evanston, IL.

During ovarian development, primordial follicles form through a process known as germ cell
nest breakdown when germ cells are encapsulated by pre-granulosa cells. In the mouse ovary,
complementary expression of the ligand Jagged1 (J1) in germ cells and the receptor Notch2 (N2)
in granulosa cells suggests a possible role for Notch signaling in mediating the cellular
interactions necessary for follicle formation. To investigate this further, we used the Cre/loxP
system to disrupt J1 specifically within germ cells (floxed mice provided by J. Lewis; VasaCre
mice provided by D. Castrillion) and N2 specifically within granulosa cells (floxed mice
provided by U. Strobl; Amhr2Cre mice provided by R. Behringer). J1 knockout (VasaCre/+,J1fl/- or
J1KO) and N2 knockout (Amhr2Cre/+,N2fl/- or N2KO) mice had a clear decrease in ovarian J1 or
N2 RNA and protein levels as detected by real-time PCR and immunohistochemistry, as
compared to controls, indicating successful disruption of each gene. Importantly, the level of N2
varied considerably among N2KO mice (reduction of 40-100% vs. WT) by Western blot.
Histological examination of J1KO and N2KO ovaries revealed the presence of numerous multi-
oocytic follicles (MOFs), with some MOFs containing as many as 13 oocytes. Both J1KO and
N2KO ovaries also contain follicles with enlarged or degenerating oocytes and exhibit an
increase in cell death within granulosa cells of follicles as determined by TUNEL staining. J1KO
mice are subfertile in comparison to heterozygous controls. Interestingly, despite similarities
with J1KO mice, initial studies indicate that female N2KO mice are fertile, with a mean litter
size similar to WT. However, 2 of the 6 N2KO females examined exhibited premature
reproductive senescence. These studies indicate an important role for J1 and N2 in the mouse
ovary. Current studies are aimed at understanding how loss of Notch signaling impacts granulosa
cell function and follicle development. Supported by NIH P01 HD021921 and NIH T32
GM12453.




                                                                                              25
Abstract # P5

Zinc maintains prophase I arrest in mouse oocytes through regulation of the MOS-MAPK
pathway. Betty Kong1, M. L. Bernhardt1, A. M. Kim1, T. V. O’Halloran2,3, T. K. Woodruff1,3
1
  Department of Obstetrics and Gynecology, Northwestern University, Feinberg School of
Medicine, Chicago, IL 60611; 2Department of Chemistry, Northwestern University, Evanston, IL
60208; 3Department of Molecular Biosciences, Northwestern University, Evanston, IL 60208.

Meiosis in mammalian females is marked by two arrest points, at prophase I and metaphase II,
which must be tightly regulated in order to produce a haploid gamete at the time of fertilization.
The transition metal zinc has emerged as a necessary and dynamic regulator of the establishment,
maintenance and exit from metaphase II arrest, but the roles of zinc during prophase I arrest are
largely unknown. The purpose of this project is to investigate and characterize the function of
zinc regulation during the first meiotic arrest. Cyclic AMP accumulation plays a pivotal role in
maintaining the first meiotic arrest at prophase I in mammalian oocytes. Oocytes cultured in the
presence of phosphodiesterase 3 inhibitors, such as milrinone, remain arrested at the prophase I
stage due to pharmacologically maintained levels of high cAMP. However, disruption of zinc
homeostasis via the addition of the heavy metal chelator N,N,N’,N’-tetrakis(2-pyridylmethyl)
ethylenediamine (TPEN) at concentrations of 10 µM or higher overrides the cAMP-maintained
arrest and causes meiotic resumption. Approximately 50% of oocytes cultured in the presence of
milrinone and TPEN undergo meiotic resumption by 9 hours. The MOS-MAPK pathway
mediates zinc-dependent prophase I arrest, as levels of MOS steadily increase prior to the onset
of meiotic resumption in TPEN treated oocytes. In contrast, MOS levels remain low in oocytes
that undergo spontaneous resumption until well after germinal vesicle breakdown has occurred.
Further, inhibition of the MOS-MAPK pathway by pharmacological inhibition or siRNA
knockdown of MOS significantly reduces TPEN-induced meiotic resumption, suggesting that the
effects of TPEN are specifically mediated through the MOS-MAPK pathway. Finally, in
contrast to prolonged zinc insufficiency, which ultimately results in telophase I arrest, early and
transient exposure of oocytes to TPEN is sufficient to induce meiotic maturation and bypass the
telophase I block, allowing the formation of developmentally competent metaphase II eggs.
These results, together with those of previous studies, establish zinc as a crucial regulator of both
meiotic arrest points, spanning the entirety of oocyte maturation. Supported by NIH Grant P01
HD021921 and the W. M. Keck Foundation Medical Research Award.




26
                                                                                 Abstract # P6

Repeated phase shifts disrupt pregnancy in the mouse. Keith Summa1,2, Martha Hotz
Vitaterna1, F.W. Turek1,2, 1Center for Sleep and Circadian Biology, Department of
Neurobiology, Northwestern University, Evanston, IL, 2Northwestern University Feinberg
School of Medicine, Chicago, IL.

The circadian clock has been linked to reproduction at many levels in mammals.
Epidemiological studies of female shift workers have reported increased rates of reproductive
problems and adverse pregnancy outcomes, although whether the cause is circadian disruption or
an associated lifestyle factor is unknown. Here we test whether environmental disruption of
circadian rhythms, using repeated shifts of the light:dark (LD) cycle, adversely affects
reproductive success in mice. Young adult female C57BL/6J (B6) mice were paired with B6
males until mating was verified by visual identification of vaginal plug formation. Females were
then randomized into one of three groups: control, phase-delay or phase-advance. Controls
remained on a stable 12-hr light:12-hr dark cycle, whereas phase-delays and phase-advances
were subjected to 6-hr delays or advances in the LD cycle every five-to-six days. The number of
copulations resulting in term pregnancies was determined. Compared to controls, reproductive
success was reduced in mice subjected to chronic circadian disruption using phase shifts of the
LD cycle, which disrupt endogenous circadian timekeeping. Phase advances have a greater
negative impact on pregnancy outcomes and require longer for circadian re-entrainment,
suggesting that the magnitude of circadian misalignment may be related to the severity of the
adverse impact on pregnancy. These results explicitly link disruptions of the light cycle to
adverse pregnancy outcomes in mammals, which may have important implications for the
reproductive health of female shift workers, women with circadian rhythm sleep disorders and/or
women with disturbed circadian rhythms for other reasons. Supported by March of Dimes
Foundation Prematurity Research Initiative Award #21-FY08-506 to FWT.




                                                                                             27
Abstract # P7

Development of ovarian cultures to rapidly test chemotherapeutic fertotoxicity in vitro and
in vivo. So-Youn Kim1, Jessina Tomas1, Vanida Ann Serna1, Marilia Cordeiro1,2, Takeshi
Kurita1, Teresa K. Woodruff1; 1Department of Obstetrics and Gynecology, Feinberg School of
Medicine, Northwestern University; 2Center of Neuroscience and Cell Biology, Coimbra,
Portugal.

Earlier detection of cancer, coupled with new and improved treatment options, has increased the
number of survivors of cancer, especially those who are diagnosed as children. As the number of
young cancer survivors increases, so does the concern for their quality of life post-treatment.
Unfortunately, off-target effects of radiation and chemotherapies can have unexpected and
lasting consequences. For young women, this is particularly worrisome, as the immature oocytes
that comprise the ovarian reserve are particularly vulnerable to the effects of radiation and drugs.
Our group has developed in vitro ovarian culture and isolated ovarian follicle culture systems in
order to advance our understanding of follicle development and the effects of cancer treatments
on the ovary, with the goal of finding new ways to restore fertility to young cancer survivors who
have been rendered sterile by their cancer treatment. A recent study suggested that the tyrosine
kinase inhibitor Gleevec® (imatinib mesylate) protects oocytes from cisplatin-induced cell death
when immature animals were treated. Options for preserving fertility in young women prior to
cancer treatment are investigational and involve removal of an ovary; even in the face of certain
sterility, this can be a difficult decision for young patients and their parents. Experiments aimed
at more rapidly testing chemotherapies as well as identifying neo-adjuvant fertoprotective agents
can be more rapidly advanced by the deployment of the in vitro follicle culture system and
kidney grafting model described here. To test our in vitro culture system and the kidney grafting
model, we first treated ovaries with Gleevec® in vitro. Here we show that Gleevec® rescues
primordial follicles by cisplatin in cultured mouse ovaries by blocking an apoptotic signaling
pathway including Bax, cPARP and Caspase 3. Further, we show that the kidney grafting model
recapitulates the in vitro results and extend our analysis to understand the effect of Gleevec.
These studies show that we have two model systems in which drug testing can occur rapidly, and
aid our identification of fertoprotective agents in the future. Supported by
NIH/NICHD Oncofertility Consoritum U54HD041857.




28
                                                                                    Abstract # P8

In vitro follicle growth (IVFG) as a novel ovarian function assay in reproductive toxicology.
Yuanming Xu1, Min Xu1, Teresa K. Woodruff1, 1Northwestern University, Feinberg School of
Medicine, Department of Obstetrics and Gynecology, Chicago, IL 6061.

Hundreds of thousand of chemical entities are produced each year by chemical and
pharmaceutical companies. Before acceptance for use, these compounds need to be
systematically evaluated for their toxicological effects, which include a screen of their
fertotoxicity. The testing of toxic effects on the reproductive function is a complex process. The
traditional in vivo approach is a time-consuming and costly process, and often only measure
overt disease phenotypes (i.e. infertility, teratogenicity, etc). There is an urgent and unmet need
to develop a rapid, in vitro-based assays that use biomarkers or upstream events that predict
adverse reproductive health effects and that help identify toxicologic pathways. We propose that
in vitro follicle growth (IVFG) can be translated into a novel in vitro ovarian function assay to
rapidly and efficiently screen the fertotoxicity of pharmaceutical environmental compounds.
Indeed, the in vitro bioassay confirms dose-response fertotoxicity of two common chemotherapy
medicines, cyclophosphamide and cisplatin. Along with visual assessment of follicular growth,
differentiation and oocyte growth and maturation by conventional light microscopy, steroid
secretion measurements allowed to study the effect of oil dispersants used in unprecedented
amounts in the Gulf of Mexico oil spill on female reproductive function. In addition, we started
to screen the fertoxicity of the NIH Clinical Collection (NCC) representing plated arrays of small
molecules that have a history of use in human clinical trials. The array that we are testing
consists almost entirely of drugs that have been in phase I-III clinical trials and have not been
represented in other arrayed collections. This in vitro ovarian assay allows us to collect
information on multiple fertility-related endpoints in the timely and cost-efficient manner. This
work was supported by the Oncofertility Consortium (NIH RL1HD058295 and NIH
PL1CA133835).




                                                                                                29
Abstract # P9

Large scale isolation of primordial follicles for transplantation within biomaterial grafts in
a mouse infertility model. Ekaterina Kniazeva¹, A.N. Hardy², L.D. Shea¹, ¹Department of
Chemical and Biological Engineering, ²Department of Surgery, Northwestern University,
Chicago, IL.

A significant challenge to applying ovarian tissue transplantation as a means of fertility
preservation to the cancer population lies in the potential to re-seed cancer cells to the patient. To
reduce the risk of disease transmission, we are investigating an approach based on isolating and
transplanting only the functional unit of the ovary, the follicular population. We propose to
employ biomaterials to encapsulate the follicles as a means to support and promote their
engraftment within the host tissue. The material can be designed to modulate the physical and
biochemical environment, and to provide a more consistent and homogeneous delivery of
follicles relative to the transplantation of ovarian tissue sections. We have refined the procedure
for large scale recovery of primordial follicles from 6 to 8-day old mice and have investigated
transplant engraftment and function within adult mouse bursa using a range of biomaterials.
Initial studies have identified a dependable procedure for isolating large numbers of immature
follicles from the donor ovarian tissue by means of combining chemical and mechanical
digestion. In vitro culture and histological analysis provided a preliminary confirmation of
follicle survival within fibrin-alginate grafts both in vitro and in vivo for up to 3 days. Further
research is targeted at evaluation of functionality of the resulting transplant within a variety of
biomaterial compositions in order to achieve desirable structural support, degradability, ease of
gel formation, biocompatibility and suitable pore size within the graft to promote follicle growth
and maturation. Supported by PL1EB008542: Biomedical Core Oncofertility Consortium
Roadmap grant from the NIH:NIBIB National Institute of Biomedical Imaging and
Bioengineering.




30
                                                                                    Abstract # P10

Exploring the effects of the Smad4 signaling pathway on Pax2 activated cells during early
mouse urogenital system development. Jerry Rhee1, Kenji Unno2, Vanida Ann Serna2, Lindsey
Marie Butler2, Philip M. Iannaccone1 and Takeshi Kurita2; 1CMRC/Northwestern Department of
Pediatrics, Developmental Biology Program; 2Center for Genetic Medicine, Northwestern
Feinberg School of Medicine, Chicago, IL.

There is very little known about the sequence of cellular interactions that generate the properly
patterned early mouse urogenital system. To investigate the requirements for the Smad4
signaling pathway in Pax2 activated cells that contribute to the construction of the mouse
reproductive system, we are examining Pax2:Cre;mTmG;Smad4fl/fl crosses from 8.5dpc to
13.5dpc. We are performing immunohistochemistry and live imaging of Pax2:Cre;mTmG and
Pax2Cre;mTmG;Smad4Δ/Δ embryos. Our preliminary examinations demonstrate that the earliest
time at which Pax2:Cre drives conversion of mTmG is at early somite stages (~8dpc) in the
posterior endoderm of the primitive streak immediately prior to hindgut invagination. The
labeled population is comprised of a mixture of cell types, including endothelial cells. Upon
induction, the activated population quickly rearranges to generate a knot-like structure ~100mm
in diameter. At 9.5dpc, the marked population constitutes the bilaterally symmetrical
intermediate mesoderm and some cells within the hindgut endoderm. At 12.5dpc, labeled cells
are especially dense at the anterior region of the gonad/mesonephros, which migrate caudally to
populate different gonadal compartments. We are currently examining the first mutant litter at
9.5dpc but preliminary observations suggest that any expressed phenotypic defect is subtle. Our
characterizations represent the highest resolution optical descriptions of the morphological
transformation affecting the early mouse urogenital system. We believe such serial descriptions
across the cell level are crucial for placing the signaling interactions in proper context to improve
rational design for developing clinical therapies. This research is supported by George M.
Eisenberg Foundation for Charities and NIH/NCI R01 CA154358.




                                                                                                  31
Abstract # P11

Down-regulation of miR-29b is essential for the pathogenesis of uterine leiomyoma. Wenan
Qiang1, Zhaojian Liu2, Vanida Ann Serna1, Yu Liu1, Stacy Druschitz1, Jian-jun Wei2, and
Takeshi Kurita1, 1Division of Reproductive Biology Research, Department of Obstetrics and
Gynecology; 2Department of Pathology; Northwestern University Feinberg School of Medicine,
Chicago, Illinois 60611.

Uterine leiomyomas (ULs) are the most common neoplasm in women of reproductive age with
an incidence of over 70%. The most notable characteristic of UL is excess extracellular matrix
(ECM), composed mainly of collagen Types I and III. Recent gene expression profiling studies
have revealed that many microRNAs are aberrantly expressed in ULs. Specifically, miR-29b is
consistently found to be down-regulated in ULs compared to normal myometrium. In vitro
studies have also shown that it can down-regulate multiple collagens. Down-regulation of miR-
29 has been correlated with an over-expression of collagen in the liver, lung, kidney and cardiac
fibrosis. One of the leading targets of miR-29b among ~1000 others was COL3A1, as shown in a
target prediction algorism (miSVR) based on the Support Vector Regression model. This
suggests that the down-regulation of miR-29b may play a role in ECM overproduction in ULs.
We established human UL cell lines that restored miR-29b expression levels to normal by
lentiviral infection to examine cell proliferation and the expression of potential miR-29b targets
including COL1A1 and 3A1. Next, the effect of miR-29b expression levels on tumor growth in
vivo with UL cell lines or primary cultures was examined by utilizing a xenograft model. The
results indicated that when miR-29b was restored to the myometrial level, the UL cells and
primary human UL cells did not form a solid tumor. Instead, they grew flat on the surface of the
host kidney. Histology indicated that an up-regulation of miR-29b inhibited collagen
accumulation. We further generated myometrial cell lines in which miR-29b was permanently
knocked down to evaluate the tumor formation of the cells. All of these results showed that
down-regulation of miR-29b is essential for UL formation, but it is not sufficient to transform
myometrium into ULs. Thus, miR-29b could be used as a potential therapeutic target for fibrotic
diseases. This research was supported by NICHD Grant R01 HD064402.




32
                                                                                Abstract # P12

The role of miR-146a in male germline stem cell self-renewal, Jessica Huszar1 and
Christopher J. Payne1, 1Department of Human Molecular Genetics, Children’s Memorial
Research Center, Chicago, IL.

Male germline stem cells (GSCs) are a subset of undifferentiated spermatogonia. They must
continually self-renew to maintain an adult stem cell pool in the mammalian testis, as well as
differentiate to initiate spermatogenesis throughout adulthood. This balance between self-
renewal and differentiation is tightly regulated. However, the mechanisms controlling this fate
decision remain poorly characterized. Here, using primary mouse testis cells enriched for either
undifferentiated or differentiated spermatogonia, we have identified a microRNA, miR-146a, that
is upregulated approximately 180-fold in undifferentiated spermatogonia. We hypothesize that
miR-146a promotes the self-renewal of GSCs by repressing genes required for differentiation. To
support this, transient overexpression of miR-146a in cultured primary GSCs decreases transcript
levels of differentiation markers. Conversely, knockdown of miR-146a increases transcript levels
of differentiation markers and decreases markers specific to undifferentiated spermatogonia.
Currently, we are focused on how these expression changes relate to GSC fate. Further studies
aim to elucidate the precise role and mechanism of miR-146a in GSC self-renewal and
differentiation.




                                                                                             33
Abstract # P13

Relating economic conditions to vasectomy and vasectomy reversal rates: a bi-institutional
study. Sherwin Zargaroff1 , Vidit Sharma1, Kunj R Sheth1, Brian V. Le1, James H. Dupree1,
Marc Goldstein2, Robert E Brannigan1; 1 Northwestern University Feinberg School of Medicine,
Department of Urology, 2 Weill Cornell Medical College of Cornell University, Department of
Urology and Institute for Reproductive Medicine and The Population Council, Center for
Biomedical Research, New York, NY.

Introduction: There has been widespread speculation in the media that vasectomy rates serve as
an indicator for economic instability. However, these claims have been largely based on single
provider anecdotes and remain unverified. Here we compare vasectomy and vasectomy reversal
rates in two institutions to national economic indicators over a 10-year period spanning two
economic recessions.
Methods: A retrospective chart review was conducted to identify the number of vasectomies per
month and vasectomy reversals per year from the interval of January 2001 to July 2011. The
incidence of these procedures were pooled from two institutions and then correlated with
national economic data as listed by the Bureau of Labor Statistics and other educational
resources.
Results: There were a total of 2612 vasectomies and 359 vasectomy reversals in both institutions
during the study period. The monthly vasectomy frequency was correlated with the following
monthly economic indicators: US leading index (r=+0.267, p=0.001), US Inflation Rate (r=-
0.192, p=0.015), S&P 500 Monthly Close (r=-0.143, p=0.054), and US Unemployment Rate (r=
+0.520, p<0.001). This indicates that vasectomy frequency is inversely related to the business
cycle, where a higher vasectomy incidence is associated with a recessionary economy. Similarly
the annual vasectomy reversal frequency was correlated with the following annual economic
indicators: US GDP growth rate (r=+0.344, p=0.105), US Gross Savings Rate (r=+0.810,
p=0.003), US Inflation Rate (r=+0.492, p=0.074), and US Unemployment Rate (r=-0.787,
p=0.004). Thus, more vasectomy reversals are performed in an expansionary economy. Annual
vasectomy and vasectomy reversal rates were inversely correlated (r=-0.432), but this
relationship did not reach statistical significance.
Conclusions: There is a clear relationship between the number of vasectomy and vasectomy
reversals performed at our institutions and the state of the economy, highlighting the importance
of financial pressures on family planning decisions. Moreover, the data indicates that vasectomy
incidence can potentially serve as a novel economic indicator. Future large-scale studies are
warranted to further explore this relationship.




34
                                                                                   Abstract # P14

Onco-fertility outcomes may differ by cancer diagnosis, Mary Ellen Pavone1, J Hirshfeld-
Cytron2, A Lawson1, K Smith1, R Kazer1, S Klock1; 1Department of Obstetrics and Gynecology,
Northwestern University, Chicago, Il; 2Department of Obstetrics and Gynecology, University of
Illinois, Chicago, IL.

We have previously shown that there are minimal differences in IVF outcomes between cancer
patients and age matched infertile controls. Here, we explore differences that exist based on
cancer diagnosis. 315 female patients expressed an initial interest in fertility preservation. Of
those, 285 had an initial appointment with a Reproductive Endocrinologist. 124 patients
expressed desire to proceed with treatment: 118 pursued embryo or oocyte cryopreservation
only, 5 pursed ovarian tissue cryopreservation only, and 1 patient had both oocyte and ovarian
tissue cryopreservation. 14 patients initially expressed interest but did not start treatment. 4
patients had their IVF cycle cancelled because of poor response. Of the 101 patients who
underwent a successful oocyte harvest, 53 had breast cancer, 26 had a hematologic malignancy,
8 had a gynecologic malignancy and 14 had another cancer diagnoses. All patients were
stimulated using an antagonist protocol. Statistics were done using parametric or non-parametric
analysis based on data distribution. Comparing the 4 groups, we found a significant difference in
age, with patients diagnosed with a gynecologic malignancy being the oldest. Prior exposure to
chemotherapy was also significantly different, with hematologic malignancies most likely to
have been exposed. Hematologic malignancies were most likely to have the longest
stimulations, with antagonist starting on the latest day; they had an overall similar number of
oocytes retrieved. There was a trend in patients with breast cancer or gynecologic malignancies
to have a greater number of oocytes retrieved and gametes frozen. Discussing fertility
preservation is an important aspect of cancer care in reproductive aged women. Despite
demonstrating an initial interest, only about 1/3 ultimately pursue onco-fertility. Further research
should be done to better understand what factors affect patient outcome and decision making.
Supported by NIH UL1DE019587, K12HD050121.




                                                                                                 35
Abstract # P15

National incidence of testicular examination in men ages 15-45: predictive factors and
barriers. KR Sheth1, Vidit Sharma1, S Zargaroff1, B Le1, JM Dupree1, RE Brannigan1,
1
  Department of Urology, Feinberg School of Medicine, Chicago, IL.

Introduction: The American Cancer Society states that "most doctors agree that examining a
man's testicles should be part of a general physical exam," and this approach might facilitate
early detection of testicular pathology. Here we use a nationwide survey to assess frequency of
both general physical exam, and more specifically, testicular exam.	
  
Methods: From 2006-2010, the CDC conducted the National Survey for Family Growth
sampling 10,403 men aged 15-45. The probabilistic and clustered design of the survey yields
national extrapolations that we have previously validated using an internal control. We used
descriptive statistics and multivariate regression models to isolate characteristics of those men
receiving testicular exams.	
  
Results: From 2006-2010 an estimated 36.9% of all men aged 15-45 in the US had a testicular
exam within the last 12 months. Multivariate regression shows testicular exams were
significantly (p<0.05) less common in immigrants (OR:0.72), older men (OR: 0.98), men lacking
a consistent health care provider (OR: 0.74), and Caucasians compared to African Americans
(OR: 0.78). The most significant predictor of receiving a testicular exam was having a physical
exam (OR: 43.0). However, of the 29,314,526 men who had a routine physical, 27.8% did not
have a testicular exam. A second regression model of men who received a routine physical exam
showed that older age (OR: 0.97), ethnicity (Caucasian vs African American, OR: 0.77), and
receiving a physical exam in a community health clinic vs a private office (OR: 0.66) were
significant (p<0.05) barriers to receiving a testicular exam. 	
  
Conclusions: In this study, the annual incidence of testicular exam in men 15-45 years of age is
only 36.9%. Surprisingly, 27.8% of men who had a routine physical did not have a testicle exam.
To facilitate the detection of gonadal pathology providers should consider conducting testicular
exams as part of a complete physical exam. 	
  
Source of Funding: none




36
                    32nd Annual Minisymposium on Reproductive Biology
                                               List of Attendees




                                      Monica Boen                               Robert Brannigan
Bernard Beckerman
                                      Feinberg School of Medicine               Urology
bernard.beckerman@gmail.com
                                      monicaboen@gmail.com                      rebrannigan@gmail.com


Rob Buerki                            Lindsey Butler                            Marshall Caraveo
Feinberg School of Medicine           Obstetrics and Gynecology                 Obstetrics and Gynecology
rbuerki@gmail.com                     lindsey-butler@northwestern.edu           marshall.caraveo@northwestern.edu


Baljash Cheema                        Michael Demczuk                           Jazzmyne Dickens
Feinberg School of Medicine           Biological Sciences, DePaul University    Obstetrics and Gynecology
baljash-cheema@fsm.northwestern.edu   mmdemczuk@gmail.com                       jazzmyne.dickens@northwestern.edu

                                      Susan Fisher
Stacy Druschitz                       Obstetrics, Gynecology and Reproductive   Georgia Gaconnet
Obstetrics and Gynecology             Sciences                                  Molecular Biosciences
s-druschitz@northwestern.edu          University of California, San Francisco   g-gaconnet@northwestern.edu
                                      sfisher@cgl.ucsf.edu

Shannon Gallagher                     Spiro Getsios                             Erv Goldberg
Children's Memorial Research Center   Dermatology                               Molecular Biosciences
s-gallagher@northwestern.edu          s-getsios@northwestern.edu                erv@northwestern.edu


Jessica Hornick                       Teresa Horton                             Dolores Huberts
Obstetrics and Gynecology             Anthropology                              Obstetrics and Gynecology
j-hornick@northwestern.edu            thorton@northwestern.edu                  dolores-huberts@fsm.northwestern.edu


Philip Iannaccone                     Toshiyuki Kakinuma                        Signe Kilen
Pediatrics                            Obstetrics and Gynecology                 Molecular Biosciences
pmi@northwestern.edu                  t-kakinuma@northwestern.edu               s-kilen@northwestern.edu


Julie Kim                             Jingjing Kipp                             Amber Kofman
Obstetrics and Gynecology             Biological Sciences, DePaul University    Pediatrics
j-kim4@northwestern.edu               jkipp@depaul.edu                          akofman@childrensmemorial.org


Pamela Kreeger
                                      Tsutomu Kune                              Takeshi Kurita
Biomedical Engineering
                                      Medicine                                  Obstetrics and Gynecology
University of Wisconsin-Madison
                                      t-kume@northwestern.edu                   t-kurita@northwestern.edu
kreeger@wisc.edu




                                                                                                            37
                    32nd Annual Minisymposium on Reproductive Biology
                                               List of Attendees




Pei-Chih Lee                           Yu Liu                                Jiajia Ma
Molecular Biosciences                  Obstetrics and Gynecology             Obstetrics and Gynecology research
p-lee@u.northwestern.edu               yliu@northwestern.edu                 jiajia-ma@northwestern.edu


Erica Marsh                            Kristen Meldi                         Navdha Mittal
Obstetrics and Gynecology              Molecular Biosciences                 Obstetrics and Gynecology
emarsh@nmff.org                        kristenmeldi2010@u.northwestern.edu   n-mittal@northwestern.edu


Diana Monsivais                        Liang Mu                              Antonia Navarro
Obstetrics and Gynecology              Pathology                             Obstetrics and Gynecology
d-monsivais@northwestern.edi           l-mu@northwestern.edu                 antonianavarro2013@u.northwestern.edu


Guadalupe Navarro                      Masanori Ono                          Christopher Payne
Endocrinology                          Obstetrics and Gynecology             Pediatrics
gnavarro@northwestern.edu              m-ono@northwestern.edu                c-payne@northwestern.edu


Rexxi Prasasya                         Emily Que                             Neena Schwartz
IGP/DGP                                Chemistry                             Neurobiology
rexxiprasasya2016@u.northwestern.edu   e-que@northwestern.edu                n-schwartz@northwestern.edu


Vanida Ann Serna                       Emily Su                              David Tagler
Obstetrics and Gynecology              Obstetrics and Gynecology             Chemical Engineering
v-serna@northwestern.edu               e-su@northwestern.edu                 david@davidtagler.com


Huanghui Tang                          Katrina-Marie Tulas                   Kenji Unno
Molecular Biosciences                  Geriatric/Oncology                    Obstetrics and Gynecology
h-tang2@northwestern.edu               katrina.tulas@northwestern.edu        k-unno@northwestern.edu


Constanza Vasquez                      Sadie Wignall                         Teerawat Wiwatpanit
Molecular Bioscience                   Molecular Biosciences                 IGP
cv.doorman@u.northwestern.edu          s-wignall@northwestern.edu            twiwatpanit@u.northwestern.edu



Teresa Woodruff                        Ju Wu                                 Min Xu
Obstetrics and Gynecology              Obstetrics and Gynecology             Obstetrics and Gynecology
tkw@northwestern.edu                   ju-wu@northwestern.edu                m-xu3@northwestern.edu




            38
                    32nd Annual Minisymposium on Reproductive Biology
                                        List of Attendees



                                Zhenghong Yu
Ping Yin                                                                Shenming Zeng
                                Molecular Pharmacology and Biological
Obstetrics and Gynecology                                               Obstetrics and Gynecology
                                Chemistry
p-yin@northwestern.edu                                                  shenming.zeng@northwestern.edu
                                zhenghong.yu@northwestern.edu


Hong Zhao
Obstetrics and Gynecology
h-zhao@northwestern.edu




                                                                                                39
                    32nd Annual Minisymposium on Reproductive Biology
                                                List of Presenters




Kelly Barry                              Jessica Huszar                      Zexu Jiao
McGarry lab                              Payne lab                           Woodruff lab
kbarry@northwestern.edu                  huszarjm@u.northwestern.edu         z-jiao@northwestern.edu



So-Youn Kim                              Ekaterina Kniazeva                  Betty Kong
Woodruff lab                             Shea lab                            Woodruff lab
so-youn-kim@northwestern.edu             ekniazeva@northwestern.edu          betkon@northwestern.edu



Monica Laronda                           Brian Le                            Yogeshwar Makanji
Kurita lab                               Robert Brannigan, Faculty Advisor   Woodruff lab
m-laronda@northwestern.edu               b-le@northwestern.edu               ymakanji@northwestern.edu



Mary Ellen Pavone                        Wenan Qiang                         Jerry Rhee
Bulun lab                                Kurita lab                          Iannaccone lab
mpavone@nmff.org                         w-qiang@northwestern.edu            jerry-rhee@northwestern.edu



Beth Sefton                              Vidit Sharma                        Robin Skory
Kim lab                                  Robert Brannigan, Faculty Advisor   Woodruff lab
elizabethsefton2008@u.northwestern.edu   vidit-sharma@fsm.northwestern.edu   r-skory@fsm.northwestern.edu



Keith Summa                              Dallas Vanorny                      Yuanming Xu
Turek lab                                Mayo lab                            Woodruff lab
ksumma@northwestern.edu                  dvanorny@u.northwestern.edu         t8j8q6@u.northwestern.edu



Sherwin Zargaroff
Robert Brannigan, Faculty Advisor
sherwin-zargaroff@fsm.northwestern.edu




            40
                                   Organizing Committee for the
                              nd
                          32 Annual Minisymposium on Reproductive Biology

Irene Lee                          Marilia Cordeiro                 Pamela Monahan, PhD                   Sherwin Zargaroff, MD
Planning Committee Chair           Graduate Student                 Postdoctoral Fellow                   Andrology Fellow
Graduate Student                   Teresa Woodruff Lab              Kelly Mayo Lab                        Robert Brannigan, Faculty Advisor
J. Julie Kim Lab                   Obstetrics & Gynecology          Molecular Biosciences                 Urology
Obstetrics & Gynecology            Feinberg School of Medicine      Weinberg College of Arts & Sciences   Feinberg School of Medicine
Feinberg School of Medicine        m-cordeiro@northwestern.edu      pamela-monahan@northwestern.edu       zargaroff@gmail.com
IreneLee2014@u.northwestern.edu




                                   The Minisymposium on Reproductive Biology is
                                             hosted and sponsored by:
                                            Center for Reproductive Science
                                                   Northwestern University
                                               2205 Tech Drive (Hogan 2-171)
                                               Evanston, IL 60208-3520, U.S.A.
                                           Phone: 847-491-5767 Fax: 847-491-5211
                                              www.northwestern.edu/research/crs


                                        Kelly Mayo, PhD                   Teresa K. Woodruff, PhD
                                            Director                         Associate Director
                                         847-491-8854                           312-503-2504

                              Fern Murdoch, PhD                                           Ingrid Miller
                          Asst Director for Operations                                Administrative Assistant
                                 847-467-2280                                             847-467-5707




                                             Special thanks to our co-sponsor:




                                           Offices of the President and Vice President for Research
  4th Illinois Symposium
 on Reproductive Sciences
                 Friday October 12, 2012
                 Northwestern Chicago Campus
 Neena B. Schwartz Lectureship Speaker
 Kenneth S. Korach, Ph.D.
 Chief, Laboratory of Reproductive and
 Developmental Toxicology, NIEHS/NIH

      Save The Dates!
Two Meetings back to back.
Same Place. Shared Sessions.
 5th Great Lakes Nuclear
   Receptor Conference
                 Fri-Sat, October 12- 13, 2012
                 Northwestern Chicago Campus
 Saturday Keynote Speaker
 Peter Tontonoz, M.D., Ph.D.
 Professor of Pathology and Laboratory Medicine
 David Geffen School of Medicine at UCLA

				
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