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111 SERUM FROM PREECLAMPTIC WOMEN INDUCES CASPASE-3ACTIVATION IN FIRST-TRIMESTER TROPHOBLAST CELLS SHAWNSTRASZEWSKI1, GIL MOR2, ROBERTO ROMERO3, DONNA NEALE4,1Yale University, Molecular, Cellular and Developmental Biology, New Haven,CT 2Yale University, Obstetrics and Gynecology, New Haven, CT 3NationalInstitute of Child Health and Human Development, Detroit, MI 4YaleUniversity, Obstetrics and Gynecology; Division of Maternal-Fetal Medicine,New Haven, CT

OBJECTIVE: Preeclampsia is associated with insufficient trophoblastinvasion, which may be a result of increased trophoblast apoptosis during thefirst trimester of pregnancy. Recently, we demonstrated that serum obtainedfrom women with preeclampsia decreased trophoblast cell viability andrendered first-trimester trophoblast cells sensitive to Fas-mediated apoptosis.In order to further investigate these results, we evaluated whether preeclampticserum activates effector caspase-3, an indicator of apoptosis, in first-trimestertrophoblast cells.

STUDY DESIGN: The H8 first-trimester trophoblast cell line was culturedin the presence of 10% serum obtained from either preeclamptic women ornormotensive pregnant controls for 48 hours. Trophoblast cell viability wasevaluated using the Cell Titer 96 assay. Activation of capase-3 was determined byWestern blot analysis.

RESULTS: Similar to our previous study, a significant decrease introphoblast cell viability was observed following treatment with serum frompreeclamptic patients in comparison to the normotensive controls. Only thepro-form of caspase-3 (p33) was detected in trophoblast cells treated with serumfrom normotensive patients. In contrast, the active forms of caspase-3 (17/19kDa) were observed in trophoblast cells treated with serum obtained frompreeclamptic patients.

CONCLUSION: The findings described in this study suggest that a linkexists between factor(s) in maternal serum and the induction of trophoblastapoptosis. The identification of the active forms of caspase-3 in trophoblast cellstreated with serum obtained from preeclamptic patients, but not from thenormotensive controls, supports the hypothesis that factors circulating inpreeclamptic women reduce trophoblast cell viability through activation of theapoptotic cascade.

112 A PROTEOMIC APPROACH FOR THE IDENTIFICATION OFBIOLOGICAL MARKERS OF PREECLAMPSIA TINNAKORN CHAIWORA-

Volume 189, Number 6Am J Obstet Gynecol

SMFM Abstracts S95

EXTREMELY ELEVATED SECOND-TRIMESTER HUMAN CHORIONICGONADOTROPIN LEVELS ARE ASSOCIATED WITH ADVERSEPREGNANCY OUTCOME NISHA VYAS1, CAROL NOREM2, DAVID WAL-TON1, EDGAR SCHOEN2, JENNIFER O’KEEFE2, ROBYN KRIEGER2,TRINH TO2, 1The Permanente Medical Group, Obstetrics and Gynecology,Oakland, CA 2The Permanente Medical Group, Genetics, Oakland, CA

OBJECTIVE: It is exceedingly rare to find an extremely elevated humanchorionic gonadotropin (hCG) level (>4.0 multiples of the median) at the timeof second-trimester maternal serum screening. The purpose of this report is toassess the clinical value of extremely elevated second-trimester hCG levels as anindicator of pregnancy complications and adverse outcomes.

STUDY DESIGN: We studied serum hCG levels in 48,028 gravidasundergoing California Expanded Alpha-Fetoprotein serum screening at 15-20weeks’ gestation during a 2-year period at a large health maintenanceorganization. Women who were screened too early, who had an indeterminatescreening result, or who had multiple gestations were excluded.

RESULTS: Of the 48,028 women, 166 (0.35%) had hCG levels greater than4.0 multiples of the median (MoM) for gestational age. The Table depicts therate of specific pregnancy outcomes occurring in women with an extremelyelevated hCG.

CONCLUSION: Pregnancies complicated by second-trimester hCG serumscreening levels greater than 4.0 MoM have a markedly higher rate of stillbirth,pregnancy-related hypertension, abruption, and second-trimester fetal loss.These findings are in sharp contrast to prior studies of elevated hCG less than 4.0MoM, which demonstrated little clinical value in predicting adverse pregnancyoutcome.

Complications and outcomes associated with extremely elevated hCG

hCG >4.0 MoM, N = 166

Stillbirth n = 5; (3%)Second-trimester loss n = 11; (7%)Hypertensive disease n = 3; (19%)Abruption n = 10; (6%)IUGR n = 7; (4%)Postpartum hemorrhage n = 8; (5%)Preterm delivery n = 12; (7%)

SOLUBLE FMS-LIKE TYROSINE KINASE 1 LEVELS ARE ELEVATED INPREECLAMPISA AND CORRELATE WITH DISEASE SEVERITY EUGENECHANG1, CHRISTOPHER ROBINSON1, DONNA JOHNSON1, MICHAELARMSTRONG1, 1Medical University of South Carolina, Obstetrics andGynecology, Charleston, SC

OBJECTIVE: Soluble Flt-1 (sFlt1) is a receptor tyrosine kinase that bindsand antagonizes the angiogenic/permeability factors, vascular endothelial(VEGF) and placental growth factor (PlGF). Excessive sFlt-1 causes hyperten-sion and proteinuria in rats. The purpose of this study was to determine whethersFlt-1 levels correlate with the severity of preeclampsia.

STUDY DESIGN: After IRB approval, blood samples from normotensivecontrols and from patients with preeclampsia were collected. Patients wereclassified with mild or severe disease based on ACOG criteria. Patients withchronic hypertension, collagen vascular disease, diabetes, and multiplegestations were excluded. Serum was stored at �708C. sFlt-1 concentration(pg/mL) was measured by ELISA. Statistical analysis was performed withANOVA and Student-t test.

RESULTS: 32 controls and 47 patients with preeclampsia were enrolled. 23patients had mild preeclampsia and 24 had severe. Mean gestational age (GA)(weeks) was similar for both controls and mildly affected patients (37.2 ± 2.0 vs35.8 ± 2.7, P = NS) but GA was more premature for severely affected patients(31.6 ± 3.0 P < 0.03) When compared to controls, sFlt-1 levels were increased inpreeclamptic patients (100.3 ± 142.0 vs 507.7 ± 441.5, P < 0.05). sFlt-1 wassignificantly higher in patients with severe preeclampsia compared to patientswith mild disease (644.9 ± 457.9 vs 344.3 ± 367.9, P < 0.05). sFlt-1 was alsosignificantly higher in patients withmore than 1 gmof proteinuria than less than1 gm (320.4 ± 408 vs 631.7 ± 406.7, P < 0.04)

CONCLUSION: sFLt-1 levels are increased in patients with preeclampsia.sFlt-1 levels correlate with the severity of disease. s-Flt-1 may antagonize thenormal effects of PlGF and VEGF.

PONGSA1, RICARDO GOMEZ1, SAMUEL EDWIN1, GIL MOR2, TIMOTHYBARDER3, ROBERTO ROMERO1, 1Perinatology Research Branch, NICHD/NIH/DHHS, Detroit, MI 2Yale University, Obstetrics and Gynecology, NewHaven, CT 3Epogen, Inc, Darien, IL

OBJECTIVE: Obstetrics is moving beyond genomics and into proteomics.The Plasma Proteome Project aims to comprehensively and quantitativelyanalyze plasma proteins to elucidate biomarkers for disease. The characteriza-tion of plasma proteins presentsmultiple challenges because of protein diversityand the large dynamic range of expression. The purpose of this study was to usea novel approach for proteomic analysis to identify differentially expressedproteins in the plasma of normal women and those with preeclampsia.

STUDY DESIGN: Two-dimensional chromatography was employed toseparate and analyze samples from normal women (n = 41) and those withpreeclampsia (n = 41). The first dimension employed high-performancechromatofocusing to identify the isoelectric point, and the second dimension,high-resolution reverse-phase high-pressure liquid chromatography to provideprotein hydrophobicity of each fraction. A 2D protein map was generated usingsoftware that displays the isoelectric point and the degree of hydrophobicity.The maps were used as pattern recognition tools to discern uniquecombinations of proteins expressed in plasma.

RESULTS: The following two partial 2D protein maps represent thedifferential plasma proteome of normal pregnant women and those withpreeclampsia. Dramatic differences between normal and preeclamptic plasmaare observed. Interface with mass spectrometry using the liquid phase outputallows protein analysis and identification.

CONCLUSION: Plasma proteomic analysis using 2D chromatographyrevealed unique differential expression in preeclampsia. This novel approachcan be used for the identification of biomarkers and patterns of biomarkerspredictive of disease.

normal (left) and PE (right)