Suppression of the aberrantly activated TNFA-NFKB pathway in the testicular niche of patients with nonobstructive azoospermia (NOA) restores spermatogonial stem cell self-renewal and expansion

Supported by: NIH/NICHD R25 HD075737, U10 HD27049, U10 HD38992, U10 HD055925, U10 HD39005, U10 HD33172, U10 HD38998, U10 HD055936, U10 HD055942, and U10 HD055944, U54HD29834, U10HD03005-0851, U10HD055925-02W1. O-93 Tuesday, October 20, 2015 11:45 AM OVARIAN TUMOR RISK IN WOMEN AFTER ASSISTED REPRODUCTIVE THERAPY (ART); 2.2 MILLION PERSON YEARS OF OBSERVATION IN GREAT BRITAIN. A. G. Sutcliffe,a C. L. Williams,a M. E. Jones,b A. J. Swerdlow,b M. C. Davies,c I. Jacobs,d B. J. Botting.a aInstitute of Child Health, University College London, London, United Kingdom; bInstitute of Cancer Research, Sutton, Surrey, United Kingdom; cReproductive Medicine Unit, University College Hospital, London, United Kingdom; dUniversity of New South Wales, Sydney, Australia. OBJECTIVE: To determine the risk of ovarian cancer, including malignant and borderline ovarian tumors, in women who have been exposed to ART. DESIGN: Records from the Human Fertilisation & Embryology Authority (HFEA) of all women who had ART in Britain between 1991-2010, were linked to the National Health Service Central Registers (NHSCR) for England, Wales and Scotland to obtain follow up for cancer outcomes, deaths and emigrations. Reporting to the HFEA is mandatory. MATERIALS AND METHODS: Cancer incidence in the cohort was stratified by age and calendar period and compared with expectations derived from annual age-specific national rates over the same period. Data were also stratified for potential mediating/moderating factors such as repeated exposures, age at first exposure, parity and subfertility diagnoses. Trends across categories were evaluated using Poisson regression. RESULTS: With 8.8 years average follow-up, 386 ovarian cancers occurred in 255,786 women. An increased risk of developing an ovarian cancer was observed in the cohort (standardized incidence ratio (SIR) 1.37; 95% CI 1.24-1.51). No increased risk was found with increasing number of cycles of ART (Ptrend¼0.80). Increasing risk was found with decreasing parity (Ptrend¼0.002), with women who had no live births by the end of treatment being at greatest risk (SIR 1.54; 95%CI 1.34-1.76). Risk was increased for women with a ‘female factor’ cause of infertility (SIR 1.62; 95%CI 1.421.84), especially endometriosis (SIR 2.35; 95%CI 1.80-3.07), but ‘male factor’ only infertility was not associated with risk (SIR 1.05; 95%CI 0.86-1.28). Younger age at starting ART carried greater cancer risk (Ptrend<0.0001). The risk of developing an ovarian cancer was strongest 0-3 years after first ART cycle (SIR 1.54; 95%CI 1.27-1.88), but no trend with duration since first treatment was observed (Ptrend¼0.30). CONCLUSIONS: Increased ovarian cancer risk was observed in this large cohort of women who had ART in Great Britain compared with national rates. The results suggest that this increase is at least partially mediated by patient factors such as low parity and endometriosis. Certain results argue against an association with ART itself (no increased risk in male factor infertility or with increasing number of cycles), but others (increased risk with decreasing age at first exposure and in the first few years after treatment) leave open the possibility that ART might affect risk. Further investigation to support these initial results will include analysis by tumor behavior and histopathological sub-type. Supported by: grants from Cancer Research UK (C36038/A11704 & A16874) and the National Institute for Health Research (NIHR-405526). We thank the Human Fertilisation and Embryology Authority, the Health and Social Care Information Centre and National Records of Scotland, for their help and support. O-94 Tuesday, October 20, 2015 12:00 PM SUPPRESSION OF THE ABERRANTLY ACTIVATED TNFA-NFKB PATHWAY IN THE TESTICULAR NICHE OF PATIENTS WITH NONOBSTRUCTIVE AZOOSPERMIA (NOA) RESTORES SPERMATOGONIAL STEM CELL SELF-RENEWAL AND EXPANSION. P. L. Yango, J. F. Smith, E. Altman, P. Logan, J. Rosen, N. D. Tran. University of California, San Francisco, San Francisco, CA. OBJECTIVE: Normal spermatogenesis is maintained by the intricate balance between self-renewal and differentiation within the spermatogonial stem cell (SSC) population primarily regulated by the testicular somatic niche. The precise mechanisms behind the pathophysiology of unexplained

FERTILITY & STERILITYÒ

NOA and potential therapeutic treatments remain to be investigated. The aims of this study are to: 1) Develop an effective in vitro model of human NOA; and 2) to utilize this model to develop a novel treatment for these patients. DESIGN: In vitro co-culture of primitive spermatogonia (SPG) and testicular somatic cells (TSCs) from subjects with normal (n¼12) spermatogenesis and with unexplained NOA (n¼12). MATERIALS AND METHODS: SPG and TSCs were isolated by fluorescence-activated cell sorting (FACS) from testicular biopsies using SSEA4 and THY1 as markers, respectively. Normal and disease SPGs were cocultured on either normal or disease specific TSCs in the presence of TNFa, TNFa inhibitor (TNFi), or conditioned media (CM). RESULTS: When normal SPG were co-cultured with either normal or NOATSCs, NOATSCs caused a 31% decrease in the rate of SSC colony formation in comparison to normal TSCs (p<0.05). In contrast, a similar rate of SSC formation was seen when either normal or NOA SPG were culture on normal TSCs. Aberrant activation of the TNFa-NFkB pathway in the NOA TSCs was detected by RNAseq and confirmed with translocation of p50 and p65 from the cytoplasm to nucleus. When normal SPG were cultured in the presence of TNFa, SSC colony formation significantly decreased by 31% and 61% in the presence of NOA and normal TSCs, respectively. In contrast, TNFi normalized the negative impact of NOA TSCs on SSC selfrenewal and expansion. Interestingly, when SPG were cultured on normal TSCs using CM collected from NOA TSCs, there was also significant decrease (35%) in SSC colony formation. However, CM collected from normal TSCs failed to rescue SSCs growing on NOA TSCs. CONCLUSIONS: The intricate interactions between somatic testicular niche and SSCs are critical to the proper balance of SSC proliferation and differentiation. Disruption of the TNFa-NFkB pathway within the normal testicular niche disrupts this balance and pushes SSCs into a prosurvival and quiescent state. Correcting the defect in this pathway allows NOA SSCs to exit their nonproliferative state and regain normal self-renewal properties. These results suggest new potential diagnostic tests and therapeutic applications for men with idiopathic NOA.

O-95 Tuesday, October 20, 2015 12:15 PM POLIDOCANOL FOAM FOR NONSURGICAL PERMANENT FEMALE CONTRACEPTION: INITIAL TRIAL IN C. Hanna,b S. Yao,b C. Bauer,c BABOONS. J. Jensen,a,b b a b O. D. Slayden. Department of OB/GYN, OHSU, Portland, OR; Oregon National Primate Research Center, Beaverton, OR; cLovelace Respiratory Research Institute, Albuquerque, NM. OBJECTIVE: Our goal is to develop a single-treatment nonsurgical method of female permanent contraception. The objective of this study was to determine if transcervical administration of polidocanol foam (PF) would prevent pregnancy in female baboons. DESIGN: Controlled nonhuman primate cohort study. MATERIALS AND METHODS: Healthy regularly cycling female baboons underwent a hysterosalpingogram for evaluation of tubal patency followed by transcervical infusion of either 20 mL of 5% PF followed by 1 mL of saline containing 100 mg doxycycline (5%/doxy; n¼5), 3% PF plus doxycycline (3%/doxy; n¼4), 3% PF with 0.01% benzalkonium chloride (3%/ BZK; n¼4), or no additional treatment (control; n¼9). All of the females received an intramuscular injection of depomedroxyprogesterone acetate (DMPA, 2 mg/kg) after the treatment. After recovery, females were socially-housed with males of proven fertility, and observed for resumption of menstrual cyclicity and evidence of mating. The primary outcomes was pregnancy within 6 months of resumption of menses. Control females that became pregnant underwent medical abortion; 5 were subsequently treated with 5% PF/Doxy/DMPA and exposed to pregnancy. PF-treated females that did not become pregnant were followed for an additional 6 months of exposure. RESULTS: All females resumed normal menstrual cycles and mating activity. After 6 months, 7/9 (78%) control females became pregnant. In contrast, fewer pregnancies occurred in treated females: 5%/doxy 1/5 (20%); 3%/doxy 1/4 (25%); 3%/BZK 2/4 (50%) (all p >.05). All pregnancies in PF-treated females were intrauterine and progressed normally to term. After 6 additional months of exposure, no other pregnancies have occurred among treated females, including the 5 control animals subsequently treated with 5% PF/doxy. Including these females, the cumulative 6 month failure in 5% PF/doxy was 10% (1/10, p < .005, Fisher’s Exact). CONCLUSIONS: A single transcervical treatment with PF prevented pregnancy in most baboons. The addition of BZK may interfere with PF

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