Human cytomegalovirus infection is associated with increased expression of the lissencephaly gene PAFAH1B1 encoding LIS1 in neural stem cells and congenitally infected brains

peer reviewedCongenital infection of the central nervous system by human cytomegalovirus (HCMV) is a leading cause of permanent sequelae, including mental retardation or neurodevelopmental abnormalities. The most severe complications include smooth brain or polymicrogyria, which are both indicative of abnormal migration of neural cells, although the underlying mechanisms remain to be determined. To gain better insight on the pathogenesis of such sequelae, we assessed the expression levels of a set of neurogenesis-related genes, using HCMV-infected human neural stem cells derived from embryonic stem cells (NSCs). Among the 84 genes tested, we found dramatically increased expression of the gene PAFAH1B1, encoding LIS1 (lissencephaly-1), in HCMV-infected versus uninfected NSCs. Consistent with these ndings, western blotting and immunouorescence analyses conrmed the increased levels of LIS1 in HCMV-infected NSCs at the protein level. We next assessed the migratory abilities of HCMV-infected NSCs and observed that infection strongly impaired the migration of NSCs, without detectable effect on their proliferation. Moreover, we observed increased immunostaining for LIS1 in brains of congenitally infected fetuses, but not in control samples, highlighting the clinical relevance of our ndings. Of note, PAFAH1B1 mutations (resulting in either haploinsufciency or gain of function) are primary causes of hereditary neurodevelopmental diseases. Notably, mutations resulting in PAFAH1B1 haploinsufciency cause classic lissencephaly. Taken together, our ndings suggest that PAFAH1B1 is a critical target of HCMV infection. They also shine a new light on the pathophysiological basis of the neurological outcomes of congenital HCMV infection, by suggesting that defective neural cell migration might contribute to the pathogenesis of the neurodevelopmental sequelae of infectio

Early evaluation of fetal heart anatomy: a prospective study from 6 to 10 weeks' gestation.

ObjectivesThe aim of the study is to evaluate the feasibility of an early cardiac examination and the sensibility in detecting congenital heart diseases between 6 and 10 weeks' gestation (WG).MethodsFrom October 2011 to January 2013, patients referred to our institution for an early scan before 11 WG were asked to participate to the study signing a written consent. A single fetal medicine specialist performed all the exams using alternatively a high frequency tranvaginal probe (6–12 MHz) or a conventional transvaginal probe (5–9 MHz) according to the distance between the probe and the fetal heart. The fetal heart anatomy has been assessed by the four chambers and great vessels views. Data were compared to the second trimester anomaly scan and autopsy.ResultsWe enrolled 733 patients and a total of 750 foetuses. Forty cases were excluded: 33 miscarriages, 3 molar pregnancies and 4 patients who refused a transvaginal scan. The 4 chambers view could be visualized in 51,4% of the cases between 6 and 8 WG, 90% of the cases between 8 and 10 WG and 97% between 10 and 11 WG. The greats vessels could be visualized in 49% of the cases before 9 WG, in 79,7% of the cases between 9 and 10 WG and in 94% of the cases between 10 and 11 WG. Seventy‐five cases were lost to the follow‐up after a normal first trimester scan. In 378 out of 635 cases, cardiac anatomy (4 chambers and great vessels) was considered normal and it was confirmed by a second trimester scan. In 7 out of 625 cases, a fetal heart abnormality was suspected. Later investigations confirmed the presence of a cardiac abnormality in 3 cases: an ectopia cordis in a Cantrell syndrome, a hypolastic left ventricle, and an atrio‐ventricular septal defect. In the remaining 4 cases, no cardiac abnormality was found.ConclusionsCardiac anatomy can be assessed very early in pregnancy, with a high sensibility in detecting congenital heart diseases and a high negative predictive value.info:eu-repo/semantics/publishe

Correlation between prenatal ultrasound and autopsy findings: a study on first trimester abortion

Objectives: The aim of the study is to evaluate discrepanciesbetween sonographic and autopsy findings following terminationof pregnancy (TOP) before 14 weeks’ gestation (WG).Methods: In a 4-year long prospective study, 48 first-trimestersTOP were performed due to fetal malformations or chromosomalabnormalities in a tertiary referral center. All patientsunderwent a vaginal delivery and a classical autopsy was performed.Ultrasound findings were compared with fetal autopsyfindings.Results: Median gestational age at termination was 13 WG and3 days. The 48 major fetal anomalies diagnosed by prenatalultrasound were: 23 chromosomal abnormalities, 7 severe neuraltube defects, 6 congenital heart diseases, 2 body stalk syndromes,1 rubella infection, 1 sickle cell anaemia, 2 kystic hygroma, 1 lowurinary tract obstruction ,2 Fetal Akinesia Deformation Sequence, 1 Cantrell Pentalogy, 2 amniotic band syndrome. All of these majorabnormalities were confirmed by fetal autopsy and no pregnancieswere terminated because of false positive ultrasound observations.Overall, a full concordance between sonographic and autopsyfindings was observed only in 50% of the cases. Discordancesconcerned the skeletal system in 22% of the cases, cranio-facialsystem in 10%, gastrointestinal system in 8%, genitourinarysystem in 8%, cardiovascular system in 4%. Concerning thecentral nervous system, specimens could not be analyzed in 30%of the cases due to their size and the tissue maceration; inthe remaining cases, discordance was observed in 6% of thecases.Conclusions: Fetal autopsy may provide additional information tothe ultrasonographic morphological evaluation of the fetus submittedto TOP due to chromosomal abnormalities or malformationsbefore 14 weeks’ gestation. The combination of prenatal ultrasoundand pathological examination can contribute to improve the qualityof perinatal care and of preconceptional counseling for subsequentpregnancies.info:eu-repo/semantics/publishe

Pulmonary Hypoplasia Associated with Congenital Heart Diseases: A Fetal Study

<div><p>Background</p><p>Abnormalities of the fetal pulmonary vasculature may affect lung morphogenesis. Postnatal studies have suggested that pulmonary hypoplasia (PH) may be associated with congenital heart diseases (CHDs).</p><p>Objective</p><p>To determine the prevalence of PH associated with CHDs, and to evaluate whether CHDs with right outflow obstruction were associated with the highest risk of lung growth impairment.</p><p>Methods</p><p>Between January 2006 and December 2010, fetuses with CHD obtained following the termination of pregnancies due to fetal abnormalities were examined in a prospective manner for the detection of heart and lung defects. CHDs were classified into five pathophysiological groups. Lung weight (LW), body weight (BW), and LW/BW ratio were analyzed for each case. The expression of CD31 and VEGF in the lung was evaluated by immunohistochemistry.</p><p>Results</p><p>Fetuses with CHDs and right outflow obstruction had significantly lower LW for a given BW, and significantly lower LW/BW ratios for a given gestational age. When defining PH as a fetal LW/BW ratio <0.015 before 28 weeks, and <0.012 after 28 weeks, PH was detected in 15 of the 119 fetuses analyzed (13%). It was significantly associated with CHD with right outflow obstruction, independently of chromosomal abnormalities and associated extracardiac abnormalities (<i>p</i><0.03). Right outflow obstruction was detected in 60% of the fetuses with CHD and PH, but in only 32% of those with CHD but no PH. In fetuses with right outflow obstruction, no difference was observed between those with PH and those without PH, in terms of the ratio of pulmonary artery diameter to aortic diameter, lung CD31 expression, or lung VEGF expression.</p><p>Conclusion</p><p>CHDs with right outflow obstruction are a significant risk factor for prenatally acquired PH. The occurrence of fetal PH is not correlated with abnormalities of the pulmonary vasculature, suggesting the involvement of perfusion-independent mechanisms.</p></div

CHD diagnosis (<i>n</i> = 119).

<p>CHD diagnosis (<i>n</i> = 119).</p

Individual PA/Ao values.

<p>Values were available for 35 fetuses with CHD and right outflow obstruction, and 45 fetuses with CHDs of other types. Horizontal bar: median value in each subgroup. Dashed horizontal line: expected normal AP/Ao value (i.e. 1.2). <i>p</i><0.0001 for comparisons between subgroups.</p

Characteristics of fetuses with CHD and PH.

<p>Characteristics of fetuses with CHD and PH.</p

Characteristics of the fetuses, assigned to two subgroups according to the association of PH with CHD.

<p>Values are medians (IQR) or <i>n</i> (% of each subgroup). BW: body weight; LW: lung weight.</p><p>*<i>P</i> value<0.05;</p><p>**<i>P</i> value<0.0001.</p

Univariate and multivariate analyses assessing the risk of PH as a function of CHD category, presence of an abnormal karyotype and the presence of a malformation likely to interfere with lung growth (IUGR, renal hypoplasia, omphalocele, thoracic dystrophy, or hydrothorax).

<p>Univariate and multivariate analyses assessing the risk of PH as a function of CHD category, presence of an abnormal karyotype and the presence of a malformation likely to interfere with lung growth (IUGR, renal hypoplasia, omphalocele, thoracic dystrophy, or hydrothorax).</p

VEGF immunohistochemistry.

<p>Original magnification×10, and ×40 magnification of the area identified by a rectangle. VEGF (brown) and counterstaining with hematoxylin. Fetuses with PH (A, C, E) were compared withfetusesof a similar gestational age without PH (B, D, F). A: Fetus with right ventricular hypoplasia and a septal defect, 18 weeks, LW/BW = 0.010; B: Fetus with pulmonary atresia and a septal defect, 16 weeks, LW/BW = 0.024; C: Fetus with tetralogy of Fallot, 22 weeks, LW/BW = 0.005; D: Fetus with atrioventricular septal defect, 17 weeks, LW/BW = 0.027; E: Fetus with pulmonary atresia and tricuspid atresia, 36 weeks, LW/BW = 0.009; F: Fetus with tetralogy of Fallot, 33 weeks, LW/BW = 0.029.</p