P1.33. VERSICAN IS A NOVEL REGULATOR FOR TROPHOBLAST EPITHELIAL- MESENCHYMAL TRANSITION AND INVASION Keyla S.N. Pires 1 , Camilla M. Gonçalves 1 , Rayane M. Botelho 1 , Ana Lucia M. Silva 1 , Liliane P.G. Tenorio 1 , Jaqueline C. Santos 1 , Eloiza L.L. Tanabe 1 , Karen S.C. Borbely 1 , Suely V. Silva 2 , Vanessa M. Freitas 2 , Alexandre U. Borbely 1 . 1 Cell Biology Laboratory, Institute of Health and Biological Sciences, Federal University of Alagoas, Macei o, Brazil; 2 Department of Cell and Developmental Biology, Institute of Biomedical Sciences, University of Sao Paulo, S~ ao Paulo, Brazil Objectives: Versican is a danger associated molecular pattern involved in numerous functions, such as migration and invasion. In the placenta, versican is involved in trophoblast differentiation and survival, with increased expression in molar pregnancies. Versican also has expression in first trimester extravillous trophoblast and in placenta accreta. As such, we aimed to investigate versican roles in trophoblast epithelial mesenchymal transition (EMT), migration and invasion, and signaling pathways involved. Methods: Versican siRNA was performed by plasmid transfection in HTR- 8/SVneo cells. Term extravillous trophoblast were used for exogenous versican addition tests. After gene silencing, cell death was analyzed by Annexin V / Propidium Iodide staining and proliferation by Ki67 staining. Cytoskeleton organization was analyzed by phalloidin, vimentin and cytokeratin-7 stainings. Migration was analyzed by wound healing assay and time-lapse videomicroscopy, and invasion through fibronectin-coated Transwell chambers. EMT and signaling pathway molecules were analyzed by flow cytometry and PCR assay. Results: Versican silencing induced reduction of F-actin stress fibers and vimentin expression. In vitro wound closure time was delayed, with reduction of cell speed, and invasion through fibronectin and EA.hy926 cells, which was restored with exogenous versican addition. Versican silencing reduced while exogenous versican increased the expression of mesenchymal markers, such as TGF-b1 and b2, Snail and ZEB-2. Versican silencing also reduced the activation of RhoA, ROCK2, AKT and mTOR. Conclusion: Versican is an important upregulator of trophoblast EMT, migration and invasion. Therefore, it could be an important target for invasive placental diseases, and its interactions deserve further in- vestigation. P1.34. EFFECTS OF ACUTE SEPSIS IN LIVER OF FEMALE AND MALE ADULT INTRAUTERINE GROWTH-RESTRICTED OFFSPRING EXPOSED TO A LOW PROTEIN DIET DURING GESTATION Anastasiya Vinokurtseva 1, 2, 3 , Reza Khazaee 2, 4, 3, 5 , Daniel B. Hardy 2, 4, 3 , Ruud Veldhuizen 2, 4, 5, 3 , Edith Arany 1, 2, 5, 6, 3 . 1 Department of Pathology and Laboratory Medicine, London, Canada; 2 Schulich School of Medicine and Dentistry, London, Canada; 3 Western University, London, Canada; 4 Department of Physiology and Pharmacology, London, Canada; 5 Lawson Health Research Institute, London, Canada; 6 Children's Health Research Institute, London, Canada Objectives: Intrauterine growth restriction (IUGR) occurs due to con- ditions of maternal malnutrition as a result of caloric restriction or to alterations in dietary composition. We and others developed an (IUGR) model by restricting proteins (LP) in the maternal diet resulting in long-term hepatic dysfunction and glucose intolerance. Humans with IUGR are predisposed to sepsis characterized by an intense inflamma- tory response or cytokine storm followed by an immune suppressive phase. One potential regulator of this proinflammatory process is the family of peroxisome proliferator activated receptors (PPARs); in particular PPARa and its downstream target gene fibroblast growth factor 21 (FGF-21). In this study we sought to examine the effects of this perinatal LP insult with and without postnatal sepsis on the liver of adult female and male offspring. Methods: Adult offspring of mothers (Wistar rats) treated with control (20% protein) or low protein (8% protein) diets were studied. At 4 months of age a single intraperitoneal injection of fecal slurry was administered and then sacrificed 6 hours later. Liver was dissected and preserved for histology and gene expression of PPARa, PPARb/d, PPARg, FGF21 and of the PPAR downstream inflammatory markers Il-1a, Il-6, Il-8 and TNFb, and the inflammasome complex (i.e. caspase-1 and NLRP3) by qPCR analysis. Results: Hematoxylin and Eosin (H&E) staining demonstrated no morphological changes in the liver. While PPARb/d did not show altered levels of gene expression in IUGR offspring, both PPARa and PPARg were reduced in sepsis in both males and females. Interestingly no changes in FGF21 were observed. Inflammatory cytokines confirmed the state of inflammation in septic animals, however showed no difference between dietary conditions. Alterations in the inflammasome were not observed in any of the groups. Conclusion: Our data showed that 6 hours of exposure to an acute sepsis did not undermine the liver inflammatory response in our adult IUGR-LP treated group P1.35. IN UTERO EXPOSURE OF D9-TETRAHYDROCANNABINOL CAUSES PLACENTAL INSUFICIENCY AND ALTERS PANCREAS DEVELOPMENT IN THE NEONATAL FEMALE OFFSPRING LEADING TO IMPAIRED GLUCOSE TOLERANCE IN ADULTHOOD Ryan Guillies 1, 2, 3 , Kendrick Lee 1, 3 , Farsad Jomani 1, 3 , Savita Dhavantari 1, 4, 3,2 , Daniel B. Hardy 5, 3 , Edith Arany 1, 4, 3, 2 . 1 Department of Pathology and Laboratory Medicine, London, Canada; 2 Lawson Research Institute, London, Canada; 3 Western University, London, Canada; 4 Schullich School of Medicine and Dentistry, London, Canada; 5 Department of Physiology and Pharmacology, London, Canada Objectives: Recent reports indicate that 1 in 5 women (18-24 years) in North America use cannabis during pregnancy. This is concerning given that D9-tetrahydrocannabinol (D9-THC), the main psychoactive compo- nent in cannabis, causes placental insufficiency and symmetrical intra- uterine growth restriction (IUGR). In vitro studies suggest that D9-THC activation of the cannabinoid receptor (CB1R) in pancreatic b-cells alters the cell cycle and stimulates apoptosis. Therefore, this study examined the effects of in utero D9-THC exposure on postnatal pancreatic function and long-term glucose homeostasis. Methods: Pregnant Wistar rats were treated with daily intraperitoneal injections of either 3 mg/kg D9-THC or vehicle from gestational day 5.5 to birth. Male and female offspring were sacrificed at post-natal day 21 or at 5 months of age. Pancreata were harvested, fixed and embedded in paraffin, sectioned and subjected to immunohistochemistry. Morphometric anal- ysis was conducted with Image J. Glucose tolerance was assessed at 5 months. Ins-1E cells were employed to determine if D9-THC exhibited direct effects on markers of b cell development. Results: Maternal exposure to D9-THC led to decreased fetal:placental ratio, symmetrical IUGR, and reduced pancreatic weight. At postnatal day 21, b-cell mass and b to a cell ratio was reduced exclusively in D9- THC-exposed female offspring. This reduction is attributed, in part, to decreases in total islet number and the number of small islets. Inter- estingly, at 5 months of age, D9-THC-exposed female offspring exhibi- ted impaired glucose tolerance with persistent reductions in b-cell mass. In Ins-1E cells, D9-THC markedly suppressed the steady state levels of Pdx-1 and insulin mRNA which are hallmarks of b-cell dif- ferentiation and function. Conclusion: In-utero D9-THC exposure leads to striking sexual dimor- phism on pancreas morphology at weaning and glucose tolerance in adulthood. While the mechanism remains to be fully elucidated, these Abstracts / Placenta 83 (2019) e1ee118 e27