Male dietary trans fat intake is inversely associated to fertilization rates

matrix scaffolds are reflective of the native tissue. These scaffolds have the potential to serve as a novel 3-dimensional platform for leiomyoma and myometrium research in the future. Supported by: National Institutes of Health Grant R21HD077479-01; National Institutes of Health Grant K12HD050121; Northwestern University Women’s Reproductive Health (WRHR) Scholar Award; Harold Amos Medical Faculty Development Award; Robert Wood Johnson Foundation O-270 Wednesday, October 21, 2015 12:30 PM LEUKEMIA INHIBITORY FACTOR (LIF) GENE POLYMORPHISM T>G (RS929271) PREDICTS IMPLANTATION AND PREGNANCY AFTER IVF/ICSI INDEPENDENT OF TP53 GENE POLYMORPHISM. J. A. Oliveira,a,b L. D. Vagnini,b A. Renzi,b G. R. Oliveira-Pelegrin,b C. G. Petersen,a,b A. L. Mauri,a,b F. C. Massaro,a,b F. Dieamant,a,b M. Cavagna,a,b,c R. Baruffi,a,b J. G. Franco, Jr.,a,b aCenter for Human Reproduction Prof. Franco Jr, Ribeirao Preto, Brazil; bPaulista Center for Diagnosis Research and Training, Ribeirao Preto, Brazil; cWomen’s Health Reference Center Perola Byington Hospital, Sao Paulo, Brazil. OBJECTIVE: Leukemia inhibitory factor (LIF) plays a critical role in embryo development and implantation. The literature provides evidence that a LIF T/G gene polymorphism (rs929271) is associated with female fertility. However, studies on LIF polymorphisms are still scarce. On the other hand The TP53 gene plays a critical role in regulating blastocyst implantation function mediate by genes involved in the TP53 pathway, including the LIF gene. Studies have demonstrated that the genotype of the TP53 codon 72 polymorphism (rs1042522; encoding arginine [Arg] or proline [Pro]) is associated with the expression of LIF. This study analyzed whether the LIF T/G gene can predict pregnancy outcomes in ART and, if so, whether this is independent of the TP53 codon 72 polymorphism. DESIGN: Prospective MATERIALS AND METHODS: A total of 411 women who had undergone cycles of IVF/ICSI were recruited. DNA was extracted from peripheral blood samples that were taken from each participant, and LIF T/G TP53 codon 72 Arg/Pro single nucleotide polymorphisms (SNP) were genotyped using real-time PCR. All procedures were performed under the same clinical/laboratory conditions. The cumulative results, including fresh and frozen cycles, were analyzed. RESULTS: Hardy-Weinberg genotype distributions of the entire sample indicated concordance between the observed and expected fre-

quencies. Characteristics such as age, infertility, etiology, number of transfers and number of transferred embryos were not significantly different (P>0.05) between the groups. The distribution of SNP genotypes at codon 72 of the TP53 gene was similar among the three genotypes of LIF SNP T/G (P>0.05).The G/G LIF genotype was associated with increased implantation and ongoing pregnancy rates after IVF/ ICSI. However, no correlation was observed between TP53 codon 72 polymorphism genotypes and clinical outcomes after IVF/ICSI.The table below summarizes the data. CONCLUSIONS: LIF SNP T/G (rs929271) appears to be a susceptibility biomarker that is capable of predicting implantation efficiency and pregnancy outcomes after IVF/ICSI. This influence appears to be independent of TP53 codon 72 polymorphism (rs1042522) genotypes. Supported by: Merck Serono grant (GFI-2014-16). The funders had no role in study design, data collection and analysis, or preparation of the abstract

NUTRITION O-271 Wednesday, October 21, 2015 11:15 AM MALE DIETARY TRANS FAT INTAKE IS INVERSELY ASSOCIATED TO FERTILIZATION RATES. M. Arvizu,a C. Tanrikut,b R. Hauser,c M. Keller,c J. E. Chavarro.c aNutrition, Harvard T.H. Chan School of Public Health, Boston, MA; bMassachusetts General Hospital, Boston, MA; cHarvard T.H. Chan School of Public Health, Boston, MA. OBJECTIVE: Saturated and trans fat intake has been related to lower sperm counts, but it is not known whether this leads to lower reproductive success among couples undergoing infertility treatment. To address this issue, we evaluated the association between dietary fat intake and fertilization rate among couples undergoing assisted reproductive technologies (ART). DESIGN: Prospective cohort study MATERIALS AND METHODS: We followed 141 men from couples presenting to an academinc fertility center whose partners underwent ART (n¼246 cycles). Diet was estimated before treatment with a validated food frequency questionnaire and outcome data were extracted from medical records. Generalized linear mixed models with random intercepts to account for multiple cycles per woman were used to evaluate the association of fat intake with fertilization rate while adjusting for male and female age, total

Fresh Cycles

GENOTYPE WOMEN’S GENOTYPES LIF T/G (RS929271) GROUPS

Implantation rate T/T n:168 T/G n:202 G/G n:41 P

GENOTYPE Implantation rate

Ongoing pregnancy rate/transfer WOMEN’S GENOTYPES Arg/Arg TP53 CODON 72 ARG/ n:198 PRO (RS1042522) GROUPS Arg/Pro n:182 Pro/Pro n:31 P

16.8%a (87/519) 17.8%b (107/601) 30.1%a,b (31/103) 0.002a 0.006b Fresh Cycles Ongoing pregnancy rate/patient 18.8% (112/595) 18.5% (100/540) 14.8% (13/88) ns

Ongoing pregnancy rate/transfer

Cumulative(Fresh+frozen/thawed) Cycles Ongoing pregnancy rate/patient

Implantation rate

Ongoing Ongoing pregnancy pregnancy rate/transfer rate/patient

19.6%a (44/225) 22.4%b (64/286) 38.8%a,b (19/49) 0.007a 0.01b

26.2%a (44/168) 31.7% (64/202) 46.3%a (19/41) 0.02a

15.9%a 18.5%a (100/630) (53/287) 16.2%b 20.2%b (121/747) (73/362) 27.0%a,b 36.7%a,b (34/126) (22/60) 0.004a 0.003a 0.005b 0.007b Cumulative (Fresh+frozen/thawed) Cycles Implantation Ongoing pregnancy Ongoing pregnancy rate rate/transfer rate/patient

31.5%a (53/168) 36.1% (73/202) 53.7%a (22/41) 0.01a

22.5% (63/280) 23.8% (56/235) 17.8% (8/45) ns

31.8% (63/198) 30.8% (56/182) 25.8% (8/31) ns

37.4% (74/198) 35.7% (65/182) 29.0% (9/31) ns

17.9% (130/728) 16.8% (111/661) 12.3% (14/114) ns

21.1% (74/350) 21.7% (65/300) 15.3% (9/59) ns

Values within columns containing the same letter were significantly different. ns: not significant.

FERTILITY & STERILITYÒ

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calories, male body mass index, race, male and female smoking status, infertility diagnosis, protocol type and dietary patterns. RESULTS: Men’s median age was 36.9 years and median total fat intake was 32% of calories/day. Fertilization rates were lowest in couples with the highest male partner intake of trans fats (median 1.20 % calories/day, IQR 1.21,1.46%). The multivariate-adjusted fertilization rates (95% Confidence Interval) in couples in increasing tertiles of men’s trans fat intake were 77% (70-84%), 78% (72-84%) and 64% (55-72%) (p, linear trend ¼ 0.05). This association was stronger in cycles with conventional insemination where the corresponding adjusted fertilization rates were 83% (69-91%), 70% (54-83%), and 47% (30-64%) (p, linear trend ¼ 0.005), and less pronounced in ICSI cycles (p, linear -trend¼0.11). CONCLUSIONS: Higher male-partner intake of trans fat was associated with lower fertilization rates in couples undergoing ART, particularly in conventional insemination cycles. Supported by: NIH grants R01ES009718, R01ES022955, P30ES000002, P30DK46200.

Ln mice (Table 1). There were no differences in the levels of IL-6, IL-10, and interferon-gamma between the groups. CONCLUSIONS: 10-week exposure to HFD causes significant reduction in primordial follicles, compromised fertility, higher pro-inflammatory cytokine levels, and increased ovarian macrophage infiltration, independent of obesity. In addition, obesity worsens the effects of HFD alone. The negative effects of HFD on primoridal follicles may be mediated by increased ovarian inflammation. To the best of our knowledge, this is the first time that HFD was found to be detrimental to fertility and ovarian function independent of obesity in an interventional study. Further studies are needed to elucidate the mechanisms behind these findings. References: 1. Norman JE. The adverse effects of obesity on reproduction. Reproduction. 2010;140:343-345. Supported by: 5T32HD040135-13A National Training Program in Reproductive Medicine (MSW) O-273 Wednesday, October 21, 2015 11:45 AM

O-272 Wednesday, October 21, 2015 11:30 AM HIGH-FAT DIET CAUSES COMPROMISED FERTILITY AND INCREASED PRO-INFLAMMATORY CYTOKINES INDEPENDENT OF OBESITY. M. E. Skaznik-Wikiel, A. J. Polotsky, J. L. McManaman. OB/GYN, University of Colorado Denver, Aurora, CO. OBJECTIVE: Female obesity is associated with ovarian dysfunction and subfertility (1). However, there are no studies to date to assess whether high-fat diet (HFD) alone (without obesity) causes reproductive dysfunction. The goal of this study was to determine if HFD impacts ovarian function, fertility, and markers of inflammation independent of obesity. DESIGN: Prospective animal study. MATERIALS AND METHODS: 5-week old mice were fed either low fat diet containing 10% fat (control group-LF-Ln) or HFD containing 60% fat. After 10 week feeding trial HFD-fed mice were divided into three groups based on body weight (BW): group 1: BW >25 g - high fat obese (HFOb), group 2: BW <22 g - high fat lean (HF-Ln), and group 3: BW of 2225 g. Approximately 1/3 of animals was included in each group. Group 3 was excluded from the study. Six animals per group were sacrificed. Ovaries were collected to assess ovarian follicle pool (primordial and growing) and to determine the degree of local inflammation (macrophage infiltration) by immunohistochemistry. Serum cytokines were measured by Milliplex Multiplex assays (Millipore EMD). The remaining 6 animals per group were kept on the same diet, and followed for breeding trials for 5 months. Statistical analysis was performed using one-way ANOVA with Bonferroni correction to account for multiple comparisons. P-value < 0.05 was considered statistically significant. RESULTS: 10-week exposure to HFD resulted in depleted primordial follicles irrespective of obese phenotype (Table 1). Numbers of growing follicles did not differ significantly between the three groups (p¼0.15). Macrophage counts revealed significant differences between LF-Ln and both HFD groups, indicating increased tissue inflammation in the ovary with HF feeding independent of obesity (Table 1). Certain serum pro-inflammatory cytokines were increased in HF-Ln and HF-Ob in comparison to LF-

HIGH FAT DIET AND AGING ARE ASSOCIATED WITH MACROPHAGE INFILTRATION IN MICE OVARIES. K. Thornton, O. Asemota, S. Jindal, M. Charron, E. Buyuk. Obstetrics and Gynecology, Albert Einstein College of Medicine/ Montefiore Medical Center, Bronx, NY. OBJECTIVE: The exact mechanism of how obesity causes reproductive dysfunction is not well understood. Since obesity is a state of chronic lowgrade inflammation characterized by macrophage infiltration into the adipose tissue and increased circulating inflammatory markers, we hypothesize that obesity and high fat diet cause macrophage infiltration in the ovaries that is associated with reproductive dysfunction. DESIGN: Prospective controlled study MATERIALS AND METHODS: Mice were subjected to dietary manipulation starting at 6w of age to develop 4 female rodent models: Group 1: C57BL/6J normal chow fed mice (NC) (N¼12), Group 2: C57BL/6J high fat diet fed mice (HF) (N¼12), Group 3: MCP-1 knockout (KO) mice (N¼6) fed HF diet, and Group 4: Agouti obese mice (N¼6) fed normal chow (A-NC). At 20w of age, 6 mice in each group were sacrificed. The remaining 6 mice in Group 1 continued their NC diet while 6 mice in Group 2 were switched to NC diet for another 12 weeks. At 32w of age, the remaining 6 mice in Groups 1 and 2 were sacrificed. At the time of sacrifice, one ovary was snap frozen in liquid nitrogen and used for RNA extraction and RT-PCR for detecting the following genes: F4/80, CD11c and CD206. Ribosomal protein 36b4 was used as loading control. Data given as units of expression  SEM and p<0.05 was considered significant. RESULTS: Mouse macrophage marker F4/80 expression is significantly increased in HF (4.95  0.74, p¼0.04), MCP-1 KO (9.6  1.34, p¼ 0.01), and Agouti (7.66  5.27, p¼ 0.03) compared to NC mice (2.47  0.48). M1 (pro-inflammatory) macrophages, measured by marker CD11c, increased significantly in HF (3.61  0.67, p¼ 0.02) with a trend towards an increase in expression in MCP-1 KO (2.8  0.41, p¼ 0.06) and Agouti (3.2  1.3, p¼0.08) compared to NC mice (1.36  0.18). M2 (anti-inflammatory) macrophages, measured by CD206, showed no significant difference amongst the four groups. F4/80 and CD11c expression significantly

Table 1. Primordial follicle number, macrophage counts, cytokine levels, and breeding trial outcome.

Overall LF-Ln vs HF-Ln LF-Ln vs HF-Ob HF-Ln vs HF-Ob LF-Ln (SEM) HF-Ln (SEM) HF-Ob (SEM) p-value p-value p-value p-value Primordial follicle numbers Macrophage counts Serum cytokines Leptin (pg/mL) IL-8 (pg/mL) GM-CSF (pg/mL) Breeding trial results Litters per mouse Number of pups per mouse Number of pups per breeding attempt

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ASRM Abstracts

1395  229 12.0  3.3

623  81 45.0  3.5

537117 54.0  10.0

0.002 <0.001

0.007 0.039

0.003 0.013

NS NS

846.2589.1 51.6  7.2 8.9  2.9

1616.61333.1 60.4  10.5 43.9  11.8

6096.34458.4 95.9  11.1 37.4  13.3

0.009 0.01 0.06

NS NS 0.06

0.013 0.015 NS

0.035 NS NS

4.7  0.2

3.7  0.4

1.8  0.8

<0.001

NS

0.001

0.03

31.3  1.8

19.8  4.1

9.8  3.4

<0.001

0.046

0.001

NS

6.2  0.4

4.1  0.7

1.9 0 .8

0.002

0.013

0.002

NS

Vol. 104, No. 3, Supplement, September 2015