PRENATAL DIAGNOSIS Prenat Diagn 2006; 26: 1115–1120. Published online 4 September 2006 in Wiley InterScience (www.interscience.wiley.com) DOI: 10.1002/pd.1562 CASE REPORT Quantitative distribution of a panel of circulating mRNA in preeclampsia versus controls Antonio Farina 1,2 *, Akihiko Sekizawa 2 , Yuditiya Purwosunu 2,3 , Nicola Rizzo 1 , Irina Banzola 1 , Manuela Concu 1 , Danila Morano 1 , Federica Giommi 1 , Maurizio Bevini 1 , Mohamad Mabrook 1 , Paolo Carinci 1 and Takashi Okai 2 1 Dept of Embryology Division of Prenatal Medicine, University of Bologna, Bologna Italy 2 Dept of Obstetrics and Gynecology, Showa University School of Medicine, Tokyo, Japan 3 Dept of Obstetrics and Gynaecology, University of Indonesia, Cipton Mangunkusumo Hospital, Jakarta, Indonesia Objective The aim of this study was to evaluate whether the quantitative distribution of a panel of circulating mRNAs from maternal whole blood of normal pregnancies is statistically different from those complicated with preeclampsia (PE) with or without intrauterine growth restriction (IUGR). Methods Maternal whole blood of six subjects with mild or severe PE with or without IUGR and 30 matched controls (1 : 5 match for gestational age) were retrospectively examined for circulating mRNA markers. Seven specific mRNA markers were identified and chosen based on previous microarray mRNA expressions performed on placental tissue from normal and PE patients. They were human placental lactogen (hPL), inhibin A, KISS-1, pregnancy-associated plasma protein-A (PAPP-A), plasminogen activator inhibitor type 1 (PAI- 1), selectin-P and vascular endothelial growth factor receptor (VEGFR), which were therefore quantified for statistical purposes. Results Median gestational age was 229 (178–283) and 232 (194–262) days for controls and cases respectively. All mRNA markers but PAPP-A, showed statistically different median values. They were hPL, inhibin A, KISS-1, PAI-1, Selectin-P, and VEGFR. Inhibin A, Selectin-P and VEGFR showed higher values than expected for controls. Instead, hPL, KISS-1 and PAI-1 values of PE patients were lower than those of controls. Selectin-P was the marker with the most aberrant difference, followed by VEGFR and KISS-1. Conclusion This preliminary analysis revealed that the median values of a panel of mRNAs from the maternal blood of PE patients were different from those of the same gestational age control group at the third trimester. If prospective studies at the second trimester could detect a related marker sufficiently able to discriminate between affected and unaffected patients and thus detect the disease before its clinical onset, then a screening project using a panel of mRNAs would be feasible. Copyright  2006 John Wiley & Sons, Ltd. KEY WORDS: preeclampsia; multivariable screening; molecular markers; real-time PCR; circulating mRNA INTRODUCTION Hypertensive disorders of pregnancy, including preecla- mpsia (PE) and pregnancy-induced hypertension, are associated with significant morbidity and mortality for both mothers and neonates (Walker, 2000). PE is a mul- tisystem disorder unique to pregnancy and causes sig- nificant morbidity and mortality worldwide. PE affects 5–8% of pregnancies (De Groot et al., 1999) and the clinical features are well recognized, characteristically manifesting in the second to third trimester. However, the underlying pathology and major pathogenic changes arise in the earlier stages of pregnancy owing to a fail- ure of trophoblast cell invasion associated with increased apoptotic and necrotic indices. Given the morbidity asso- ciated with PE, an enormous variety of biomolecules *Correspondence to: Antonio Farina, Department of Embryol- ogy, Division of Prenatal Medicine, Via Belmeloro 8, 40126 Bologna, Italy. E-mail: antonio.farina@unibo.it have been studied to detect those showing evidence of alteration in the maternal circulation during early pregnancy, prior to the presentation of clinical symp- toms (Myatt and Miodovnik, 1999). The identifica- tion of reliable screening markers would permit major improvements in obstetric care through better target- ing of ante partum surveillance. A great number of possible predictive markers have been tested, includ- ing placental factors (Lambert-Messerlian et al., 2000), abnormal fibrolytic activity (Ballegeer et al., 1989; Hal- ligan et al., 1994), markers of endothelial dysfunction (Bilban et al., 2004), markers of oxidative stress (Cotter et al., 2001) and markers of abnormal trophoblast inva- sion (Clausen et al., 2000; Bosio et al., 2001a). More recently, new findings concerning cell-free fetal and maternal DNA and RNA circulating in plasma/serum of pregnant women have opened up new methods for predicting PE (Leung et al., 2001; Farina et al., 2004a; Sekizawa et al., 2004). In fact, aberrant quantitative expression of some circulating placenta-specific mRNAs Copyright  2006 John Wiley & Sons, Ltd. Received: 7 February 2006 Revised: 10 June 2006 Accepted: 14 July 2006 Published online: 4 September 2006