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Tumour-targeted adenoviral vectors show enhanced efficacy against uterine fibroids in a murine model
O-260 Wednesday, October 22, 2014 12:45 PM TUMOUR-TARGETED ADENOVIRAL VECTORS SHOW ENHANCED EFFICACY AGAINST UTERINE FIBROIDS IN A MURINE MODEL. S. A. Mohamed,a,c S. Shalaby,b,c A. Laknaur,c S. Nair,c A. Al-Hendy.c aMasoura Medical School, Mansoura, Dakahleyia, Egypt; bTanta Medical School, Tanta, Gharbeyia, Egypt; cGeorgia Regents University, Medical College of Georgia, Augusta, GA. OBJECTIVE: Fibroids are steroid hormone-dependent uterine tumors with no effective medicinal therapy at the moment; and thus hysterectomy is the mainstay of treatment. Our laboratory has been developing selective targeted gene therapy strategies as a potentially viable alternative localized therapy for uterine fibroids.(1) The question in our study is whether targeted adenovirus vector, Ad-SSTR-RGD-TK (Adenovirus human somatostatin receptor subtype 2- arginine, glycine and aspartate thymidine kinase), as a suicidal gene therapy targeted to fibroids lesions in nude mice is indeed more effective than Ad-TK. DESIGN: in vivo study. MATERIALS AND METHODS: Sixteen nude mice implanted with estrogen pellets were subcutaneously injected with rat ELT3 fibroid cells (10 million/mouse) in the right flank . After the tumors were palpable, a single direct intra tumor injection of Ad-SSTR-RGD, untargeted Ad-TK, or AdLacZ was given followed by daily intra-peritoneal injection of GCV (5 days). Animals were evaluated regularly and tumors measured weekly. Samples from tumors and other body organs were collected at day 20 and day 40 of treatment to assess the efficacy of the modified vector. Evaluation of tumor reduction on day 20 and day 30, as well as apoptosis and proliferation markers was performed in our laboratory. RESULTS: Western blot analysis showed that, in comparison with the control Ad-LacZ vector, Ad-TK caused a 44% reduction in the expression of cell proliferation protein PCNA (Proliferating Cell Nuclear Antigen), and AdSSTR-RGD-TK/GCV more effectively mediated a 60% reduction in the expression of PCNA. Regarding the activity of the apoptotic protein product of cleavage of PARP1 (Poly ADP Ribosyl Polymerase1) we found that AdSSTR-RGD-TK/GCV induced apoptosis 6 folds compared to Ad-TK (P<0.05). Immunohistochemistry using Trichrome staining showed dramatic decrease in cellularity in fibroid lesions treated with Ad-SSTR-RGD group compared to Ad-TK group. Additionally, IHC using anti PCNA antibodies showed that Ad-TK & Ad-SSTR-RGD-TK/GCV effectively reduced expression of PCNA by 70% and 90% respectively, compared to Ad-LacZ control. CONCLUSION: Fibroid-specific targeting strategies for adenoviral vectors enhance their ability to reduce cell proliferation and induce apoptosis in a fibroid mouse model. Adenoviral-based gene therapy may be a viable localized non-surgical option for effective treatment of uterine fibroid. Supported by: NIH/NICHD R01 HD046228. ENVIRONMENT AND REPRODUCTION O-261 Wednesday, October 22, 2014 11:15 AM URINARY CONCENTRATIONS OF BENZOPHENONE-TYPE UV FILTERS AND COUPLE FECUNDITY. G M. Buck Louis,a R. Sundaram,a K. Sapra,a J. Maisog,b K. Kannan.c aDivision of Intramural Population Health, NICHD, Rockville, MD; bDivision of Intramural Population Health, Glotech, Inc., Rockville, MD; cWadsworth Center, NYS Department of Health, Albany, NY. OBJECTIVE: Recently, concern has arisen about the potential toxicity of benzophenone-type ultra violet (UV) filters, chemicals added to sunscreen and personal care products, that are reported to have estrogenic and antiandrogenic in vivo and in vitro activity. DESIGN: Prospective cohort with longitudinal followup through one year of trying. MATERIALS AND METHODS: 501 couples stopping contraception to become pregnant were recruited from 16 counties in Michigan and Texas. Each partner of the couple provided urines following the baseline interview and each completed daily journals. Women utilized home fertility monitors to time intercourse relative to ovulation, and digital home pregnancy tests starting on the day of expected menstruation. Five UV filters were quantified (ng/mL) using triple-quadrupole mass spectrometry with ongoing quality control procedures: BP-1 (2,4-dihydroxybenzophenone), BP-2 (2,2’,4,4’-tetrahydroxybenzophenone), BP-3 (2-hydroxy-4-methoxybenzophenone),
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BP-8 (2,2’-dihydroxy-4-methoxybenzophenone), and 4-OH-BP (4-hydroxybenzophenone). Adjusted fecundability odds ratio (FORs) and 95% confidence intervals (CIs) were estimated for each UV filter dichotomized at the 75th percentile and adjusting for age, creatinine, body mass index, serum cotinine, and site while accounting for time off contraception. Separate models were run for each UV filter and partner, but final models included both partners’ concentrations given their low correlations (r¼%0.17). RESULTS: For male partners, BP-2 was significantly associated with reduced fecundity (FOR¼0.69; 95% CI¼0.50, 0.95). In models adjusting for both partners’ concentrations, BP-2 remained significantly associated with reduced fecundity for males (FOR¼0.70; 95% CI¼0.50, 0.97). When modeled as continuous concentrations however, the directionality for BP-2 remained but no longer retained significance. No UV-filters were significantly associated with fecundity when assessing females’ urinary concentrations. CONCLUSION: To our knowledge, these are the first human evidence suggesting that male partners’ exposures to select UV filters may diminish fecundity resulting in a longer TTP. If the findings are corroborated, reducing exposure to these estrogenic compounds may help improve fecundity. Supported by: NICHD Intramural funding (contracts #N01-HD-3-3355; N01-HD-3-3356; NOH-HD-3-3358; HHSN27500001). O-262 Wednesday, October 22, 2014 11:30 AM PERFLUORINATED CHEMICALS AND TIME TO PREGNANCY: FURTHER EVIDENCE FROM THE MIREC STUDY, A CANADIAN PREGNANCY AND BIRTH COHORT. M. P. Velez,a T. E. Arbuckle,b W. D. Fraser.a aUniversity of Montreal, Montreal, QC, Canada; bPopulation Studies Division, Healthy Environments and Consumer Safety Branch, Health Canada, Ottawa, ON, Canada. OBJECTIVE: To evaluate the impact of perfluorooctane sulfonate (PFOS), perfluorooctanoate (PFOA), and Perfluorohexane sulfonate (PFHxS) on Time to Pregnancy (TTP). DESIGN: Pregnancy-based retrospective TTP study. MATERIALS AND METHODS: The Maternal-Infant Research on Environmental Chemicals (MIREC) Study recruited 2000 women before 14 weeks of pregnancy in 10 cities across Canada between 2008 and 2011.1 Questionnaire data and biospecimens were analyzed from 1614 participants. Pregnancies following birth control failure or fertility treatment were excluded. TTP was defined as the self-reported number of months of unprotected intercourse needed to become pregnant. Plasma concentrations of PFOA, PFOS, and PFHxS measured in the first trimester were considered as a proxy of preconception exposure. Fecundability odds ratios (FORs) were estimated using Cox proportional hazard models for discrete time. FORs < 1 indicate decreased fecundability and a longer TTP. Each chemical concentration (ng/ml) was log transformed and divided by its standard deviation. Potential confounders were maternal age, education, obesity, and smoking. In our causal model, parity did not satisfy the criteria of confounding, therefore it was not included in the adjusted models. RESULTS: The probabilities of pregnancy at months 1, 6 and 12 were 0.45 (95% CI 0.43-0.48), 0.85 (95% CI 0.84-0.87), and 0.91 (95% CI 0.90-0.92), respectively. The mean maternal age was 32.6 (SD 4.9) years. Half of the women had at least one prior pregnancy with live birth, and about 15% were obese. Lower education, obesity and no previous pregnancies were associated with longer TTP. Maternal or paternal active smoking, country of birth, and household income were not associated with TTP. The geometric mean (ng/ml) of PFOA, PFOS, and PFHxS was 1.64 (95% CI 1.59-1.69), 4.57 (95% CI 4.44-4.70), and 0.99 (95% CI 0.95-1.03), respectively. Crude FORs were 0.92 (95% CI 0.83-1.02) for PFOA, 1.01 (95% CI 0.73-1.41) for PFOS, and 0.98 (95% CI 0.88-1.10) for PFHxS. After adjustment by maternal age, education, and obesity, PFOA was associated with a 12% reduction in fecundability per one standard deviation increase in log-transformed concentrations (FOR¼ 0.88; 95% CI 0.79-0.98; p¼0.02), while no significant association was observed for PFOS and PFHxS. CONCLUSION: Our results add to the evidence that exposure to PFOA, even at lower levels than previously reported, may reduce fecundability. The mechanisms involved in this endocrine disrupting effect need to be assessed. Supported by: The MIREC Study is Supported by the Chemicals Management Plan of Health Canada, the Canadian Institutes for Health Research (CIHR, grant # MOP – 81285) and the Ontario Ministry of the Environment. M.P. Velez is Supported by a CIHR Fellowship Award. W.D Fraser is Supported by a CIHR Canada Research Chair.
Vol. 102, No. 3, Supplement, September 2014
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BP-8 (2,2’-dihydroxy-4-methoxybenzophenone), and 4-OH-BP (4-hydroxybenzophenone). Adjusted fecundability odds ratio (FORs) and 95% confidence intervals (CIs) were estimated for each UV filter dichotomized at the 75th percentile and adjusting for age, creatinine, body mass index, serum cotinine, and site while accounting for time off contraception. Separate models were run for each UV filter and partner, but final models included both partners’ concentrations given their low correlations (r¼%0.17). RESULTS: For male partners, BP-2 was significantly associated with reduced fecundity (FOR¼0.69; 95% CI¼0.50, 0.95). In models adjusting for both partners’ concentrations, BP-2 remained significantly associated with reduced fecundity for males (FOR¼0.70; 95% CI¼0.50, 0.97). When modeled as continuous concentrations however, the directionality for BP-2 remained but no longer retained significance. No UV-filters were significantly associated with fecundity when assessing females’ urinary concentrations. CONCLUSION: To our knowledge, these are the first human evidence suggesting that male partners’ exposures to select UV filters may diminish fecundity resulting in a longer TTP. If the findings are corroborated, reducing exposure to these estrogenic compounds may help improve fecundity. Supported by: NICHD Intramural funding (contracts #N01-HD-3-3355; N01-HD-3-3356; NOH-HD-3-3358; HHSN27500001). O-262 Wednesday, October 22, 2014 11:30 AM PERFLUORINATED CHEMICALS AND TIME TO PREGNANCY: FURTHER EVIDENCE FROM THE MIREC STUDY, A CANADIAN PREGNANCY AND BIRTH COHORT. M. P. Velez,a T. E. Arbuckle,b W. D. Fraser.a aUniversity of Montreal, Montreal, QC, Canada; bPopulation Studies Division, Healthy Environments and Consumer Safety Branch, Health Canada, Ottawa, ON, Canada. OBJECTIVE: To evaluate the impact of perfluorooctane sulfonate (PFOS), perfluorooctanoate (PFOA), and Perfluorohexane sulfonate (PFHxS) on Time to Pregnancy (TTP). DESIGN: Pregnancy-based retrospective TTP study. MATERIALS AND METHODS: The Maternal-Infant Research on Environmental Chemicals (MIREC) Study recruited 2000 women before 14 weeks of pregnancy in 10 cities across Canada between 2008 and 2011.1 Questionnaire data and biospecimens were analyzed from 1614 participants. Pregnancies following birth control failure or fertility treatment were excluded. TTP was defined as the self-reported number of months of unprotected intercourse needed to become pregnant. Plasma concentrations of PFOA, PFOS, and PFHxS measured in the first trimester were considered as a proxy of preconception exposure. Fecundability odds ratios (FORs) were estimated using Cox proportional hazard models for discrete time. FORs < 1 indicate decreased fecundability and a longer TTP. Each chemical concentration (ng/ml) was log transformed and divided by its standard deviation. Potential confounders were maternal age, education, obesity, and smoking. In our causal model, parity did not satisfy the criteria of confounding, therefore it was not included in the adjusted models. RESULTS: The probabilities of pregnancy at months 1, 6 and 12 were 0.45 (95% CI 0.43-0.48), 0.85 (95% CI 0.84-0.87), and 0.91 (95% CI 0.90-0.92), respectively. The mean maternal age was 32.6 (SD 4.9) years. Half of the women had at least one prior pregnancy with live birth, and about 15% were obese. Lower education, obesity and no previous pregnancies were associated with longer TTP. Maternal or paternal active smoking, country of birth, and household income were not associated with TTP. The geometric mean (ng/ml) of PFOA, PFOS, and PFHxS was 1.64 (95% CI 1.59-1.69), 4.57 (95% CI 4.44-4.70), and 0.99 (95% CI 0.95-1.03), respectively. Crude FORs were 0.92 (95% CI 0.83-1.02) for PFOA, 1.01 (95% CI 0.73-1.41) for PFOS, and 0.98 (95% CI 0.88-1.10) for PFHxS. After adjustment by maternal age, education, and obesity, PFOA was associated with a 12% reduction in fecundability per one standard deviation increase in log-transformed concentrations (FOR¼ 0.88; 95% CI 0.79-0.98; p¼0.02), while no significant association was observed for PFOS and PFHxS. CONCLUSION: Our results add to the evidence that exposure to PFOA, even at lower levels than previously reported, may reduce fecundability. The mechanisms involved in this endocrine disrupting effect need to be assessed. Supported by: The MIREC Study is Supported by the Chemicals Management Plan of Health Canada, the Canadian Institutes for Health Research (CIHR, grant # MOP – 81285) and the Ontario Ministry of the Environment. M.P. Velez is Supported by a CIHR Fellowship Award. W.D Fraser is Supported by a CIHR Canada Research Chair.
Vol. 102, No. 3, Supplement, September 2014