608: Senescence in genotoxic amniocytes and stressed trophoblasts

608 Senescence in genotoxic amniocytes and stressed trophoblasts Tal Biron-Shental, Moshe Fejgin, Yehudit Sharon, Dvora Kidron, Aliza Amiel Meir Medical Center, Sackler School of Medicine, Tel Aviv University, Obstetrics and Gynecology, Kfar Saba, Israel, Meir Medical Center, Tel Aviv University, Genetic Institute, Kfar Saba, Israel, Bar Ilan University, Faculty of Life Science, Ramat Gan, Israel, Meir Medical Center, Sackler School of Medicine, Tel Aviv University, Pathology, Kfar Saba, Israel OBJECTIVE: Cellular senescence is a state of metabolic and cell cycle arrest evoked by exogenous or endogenous stressors, especially genotoxic stress. Some cases of senescence are characterized by formation of senescence-associated heterochromatin foci (SAHF). Short, dysfunctional telomeres lead to senescence. We have previously reported short telomeres in aneuploidy, intrauterine hypoxia (which causes intrauterine fetal growth restriction [IUGR]), and in uncontrolled gestational diabetes mellitus. We studied senescence in amniocytes and placentas from aneuploidic fetuses, in trophoblasts from IUGR fetuses, and from diabetic placentas, by assessing SAHF. STUDY DESIGN: Using DAPI staining, we counted the percentage of cells with increased nuclear fragmentation (SAHF). We assessed SAHF expression in trophoblasts from placental biopsies (n 7) and in amniocytes (n 10) from known trisomy-21 fetuses and compared the results to control placentas (n 6) and to amniocytes (n 10) from fetuses with a known normal karyotype. We also assessed the presence of SAHF in trophoblasts from pregnancies complicated by IUGR (n 12) and from pregnancies complicated by uncontrolled diabetes mellitus [DM] (n 10) and compared them to gestational-agematched controls from uncomplicated pregnancies (n 12). RESULTS: A significantly higher rate of cells with SAHF was found in trisomy-21 placentas and amniocytes, as well as in IUGR and in trophoblasts from pregnancies with uncontrolled diabetes compared to the control samples (Figure). CONCLUSION: Increased SAHF formation in amniocytes and in trophoblasts that were previously found to express shorter telomeres emphasizes the correlation between short telomeres and senescence. These findings may be linked to other genetic instability parameters previously described in trisomy-21 and help explain intrauterine programming in pregnancies with IUGR and uncontrolled diabetes mellitus. 609 Insulin growth factor 1 (IGF-1) therapy restored IUGR induced alteration in pancreatic tissues Khaled Omar, Tarek Alsaied, Ahmed Lababidi, Helen Jones, Swathi Balaji, Sundeeb Keswani, Timothy Crombleholme, Mounira Habli Cincinnati Childrens Hospital medical center, Center for molecular fetal therapy, Cincinnati, OH, Cincinnati Childrens Hospital medical center, Pediatrics, Cincinnati, OH, Colorado Childrens Hospital medical center, Pediatric surgery, Aurora, CO OBJECTIVE: We demonstrated that intraplacental gene therapy with adenoviral insulin growth factor (Ad-hIGF1) corrects impaired birthweight and glucose regulatory response in a mouse model of placental insufficiency. Evidence from diabetic patients and Murine IUGR offspring demonstrates decreased area and mass of islets and beta cells. We hypothesized that one of the mechanisms of AdhIGf1 induced attenuation in IUGR related adult onset diabetes is by restoring pancreatic mass, beta and alpha cell ratio. STUDY DESIGN: Laparotomy was performed on pregnant C57BL/6J mice at gestational day 18 and pups were divided into 3 groups (n 3). Control: Sham operated; IUGR: by ligation of a branch of mesenteric uterine artery; IGF1-Treated: intra placental injection of Ad hIGF1 after ligation. Pups were delivered on day 20, cross-fostered to CD1 mice. At 32 weeks, Pancreas of the off springs were weighed, and histologically processed and whole mounted. After every 20 section, a 5 um consecutive primary and reference sections were obtained for the entire pancreas. Immunohistochemistry staining was performed for Beta (anti-insulin) and Alpha (anti-glucagon) cells, and area was quantified (design-based stereology). Data were analysed using ANOVA. RESULTS: Male IUGR demonstrated 22% significant reduction in pancreatic mass (230.6 5.8 vs 296 5.7 vs 290 10 mg, p 0.01), 84 % reduction in Islets/Pancreas percentage area (2.5 1.1% vs 14.8 2.6% vs 16 3.7%, p 0.02), 80% in Beta cell/Alpha cell percentage area and 12% in Beta cell/ Islets percentage area (Table) as compared to SHAM, which was restored to normal with IGF1 treatment. Total area of pancreas, Islets, beta and alpha cells did not show any significant differences among 3 groups. CONCLUSION: Intra-placental gene transfer of Ad-hIGF1 restored IUGR-induced alterations in pancreatic mass, beta and alpha cell ratio. These changes may represent a potential mechanism of fetal reprogramming by which (AdhIGF1) may attenuate fetal predisposition to adult onset Diabetes. Algorithm for stepwise, contingent Down syndrome screening www.AJOG.org Academic Issues, Antepartum Fetal, Clinical Ob, Fetus, Genetics, Hypertension, Med-Surg-Diseases, Operative Ob, U/S Poster Session IV