Placental Factor Research Articles

S.22.03 The impact of the early life environment on stress resilience in adulthood

A greater understanding of cerebrovascular health and disease requires the consideration of recent neuroscience advances concerning neuroplasticity in the context of classical developmental neurology principles. Consideration of the ontogenetic interplay of nature and nurture influencing brain development during prenatal and early postnatal time periods should consider the concept of the developmental origins of neurological health and disease. Adaptive and maladaptive effects of neuroplasticity require a systems biology approach integrating molecular, receptor, cellular, neural network, and behavioral perspectives, culminating in the structural and functional cerebrovascular phenotypes that express health or disease across the lifespan. Cognizance of the interrelationships among maternal, placental, fetal, and neonatal factors requires an interdisciplinary appreciation of genetic/epigenetic forces of neuroplasticity during early life that incrementally influence cerebrovascular health or disease throughout childhood and adulthood. Knowledge of the systemic effects of multiorgan function on cerebrovascular development further broadens the systems biology approach to general plasticity of the individual as a whole organism. Short- and long-term consequences of the positive and negative effects of neuroplasticity must consider ongoing gene–environment interactions with maturation and aging, superimposed on earlier fetal/neonatal experiences that sustain neurological health or contribute to disease during childhood and adulthood.

A prospective study on the effects and prevalence of maternal, fetal and placental risk factors on low birth weight neonates

Background: Maternal, foetal and placental risk factors have a causative effect in the prematurity and failure to thrive in the early infantile period. Timely diagnosis helps in the anticipation of complications specific to risk factors and managing by impairing their harmful effects on the growth and development of the baby.Methods: The study was conducted prospectively in Department of Paediatrics, Rajah Muthiah Medical College and Hospital, Chidambaram from January 2018 to June 2018. 361 Low birth weight babies were included. Neonates (n=361) weighing less than 2.5 kilograms with parental informed consent are included, whereas those with severe congenital anomaly, systemic disease or infection were excluded from the study. Various known maternal, placental and foetal risk factors are observed during the course of the study. The gestational age of the neonates was estimated using modified Dubowitz scoring. Other factors like mode of delivery of the babies were also noted. Statistical analysis (descriptive statistics) was done by using Microsoft Word 2010.Results: Out of 1040 babies born from January to July 2018, 341 babies weight less than 2.5 kg. Out of which 162 (44.88%) babies were born preterm. Of the full term births 52% were low birth weights The risk factors of the neonates are not mutually exclusive. In the present study 28 maternal risk factors were studied. The prevalence of maternal, fetal and placental risk factors was 85.32%, 74.79% and 6.65% respectively.Conclusions: Period prevalence of low birth weight babies and prematurity is 34.71% and 15.58% respectively. Risk factors are not independent to each other with maternal and foetal risk factors have almost have an effect on nearly three fourths of low birth weight neonates.

894: Can we predict yield of Chromosomal Microarray in intrauterine growth restriction according to clinical parameters?

Intrauterine growth restriction (IUGR) is a condition in which the fetus fails to achieve its full growth potential. There are various etiologies for IUGR including placental factors, infections, maternal chronic conditions and genetic causes. We aimed to evaluate the yield of Chromosomal Microarray Analysis in different clinical scenarios as well as in pregnancies in which short lone bones were the major finding. Our cohort included 246 cases of pregnancies. In 182 cases IUGR was the prominent feature while in 64 cases fetal short long bones were the prominent feature. CMA analysis was performed for all cases between January 2016 to March 2018. Data were retrieved from the Israeli national database and from patient medical records. In 130 cases data regarding the clinical and familial background and prenatal work-up performed during the pregnancy were available for sub analysis. Data collection is ongoing The mean gestational age at diagnosis was 31.5± 9.4 weeks. The mean measured percentile was 3.8 ± 2.4. In 14 out of 246 cases (5.7%) CMA result was pathogenic/likely pathogenic. In 3 cases (1.2%) a microdeletion/duplication with low penetrance was detected and in 23 cases (9.3%) variables of unknown clinical significance were detected. There were 2 cases of trisomy 18, one case of trisomy 21, one case of triploidy and 10 cases of microdeletions/microduplications 5 of which were detectable by karyotype. In pregnancies in which short long bones were the prominent finding, CMA was positive in 4 cases (6.25% ). In IUGR only cases there were 10 pathological CMA results (5.5%). The yield of CMA was significantly higher for cases with pathological echocardiogram (p=0.003).And was close to reaching clinical significance for IUGR pregnancies, in which the lowest measured percentile was below 1 (p=0.057). There was no significant difference for other parameters such as maternal illness, familial background of genetic conditions, obstetrics history, and prenatal work-up. The yield of CMA for IUGR is 5.7% and is higher in cases with pathological echocardiogram results and in pregnancies in which measured percentiles were below the 1. Other parameters did not reach clinical significance; however, data collection is ongoing, hence the power of such analysis will improve and may reflect additional contributing parameters. The incremental yield of CMA over karyotyping in our cohort karyotype was 64.28%.View Large Image Figure ViewerDownload Hi-res image Download (PPT)

THE ROLE OF PLACENTAL DISORDERS AMONG RISK FACTORS IN THE FORMATION OF ISOLATED HYPOSPADIAS

Aim of the study. The incidence of hypospadias increased by 2-3 times within the last 50 years. Changes in the frequency occurred mostly due to the gain of the distal isolated forms. The etiology of the majority of cases of the isolated hypospadias isn’t clear. The study of the origin hypospadias is at a stage of the search and assessment of the value of risk factors. The multifactorial pathology is supposed to be caused by changes in an organism of the mother, a placenta and environment factors. Our research is devoted to the assessment of placental factors in the formation of hypospadias. Material and Methods. We have performed a retrospective questionnaire among parents of 99 children with the isolated forms of hypospadias. The special attention was paid to the collecting of the family anamnesis, including data on diseases of reproductive organs, both at parents and at the immediate family in the male line and also data on a occupation, addictions (smoking) of parents. Information was also collected about the peculiarities of the course of the prenatal period and the outcome of childbirth. On the basis of the obtained data, an assessment of the influence of various “parental” factors on the formation of the isolated hypospadias forms in their sons is given. Results. In 4 out of 99 cases, the origin of isolated hypospadias has been caused by genetic deviations. Primary diseases of reproductive organs in parents have been noted as follow: in fathers in 18 of 95 cases, and in mothers in 12 cases. Most often the placenta pathology in the first trimester of pregnancy was shown by the threat of a spontaneous miscarriage (in 35 cases from 95). Placental disturbances during all pregnancy were accompanied by premature births in 25 cases. In 22 cases placental disorders were shown by a fetus arrest of development. Conclusions. In our research placental disturbances were the most frequent risk factors in the formation of the isolated hypospadias.

Maternal exercise and growth restriction in rats alters placental angiogenic factors and blood space area in a sex-specific manner

Fetal growth and development are dependent on adequate placental nutrient transfer. The surface area of the placental villous network is a key determinant of nutrient exchange, which is regulated by vasculogenic and angiogenic factors. These factors are altered by intrauterine growth restriction (IUGR) and maternal obesity in both the first (F1) and second (F2) generations. We investigated the impact of endurance exercise in IUGR dams fed a High-fat diet on placental vasculogenesis and angiogenesis. Uteroplacental insufficiency (Restricted) or sham (Control) surgery was induced on embryonic day (E) 18 in Wistar-Kyoto rats. F1 offspring were fed a Chow or High-fat diet from weaning, and at 16 weeks were further allocated an exercise protocol; Sedentary, Exercised prior to and during pregnancy (Exercise), or Exercised during pregnancy only (PregEx). Females were mated (20 weeks) and F2 placentae collected at E20. Maternal Restriction, High-fat feeding and Exercise had a minimal impact on placental regulators of vasculogenesis and angiogenesis. However, Restriction increased placental labyrinth tissue area in Chow-fed dams. PregEx induced overt adaptations, including increased VEGFA and decreased PLGF protein expression, and reduced blood space area. These alterations were sex-dependent and associated with alterations in miRNA27a, a known regulator of VEGF translation. These data highlight that maternal exercise initiated during pregnancy (PregEx) causes alterations in placental vasculogenesis and angiogenesis in a sex-dependent manner, with minimal Restriction and maternal diet effects. However, further investigation is required to determine if these adaptations are beneficial or harmful for maternal and fetoplacental outcomes.

A Prospective study on the effects & prevalence of maternal, fetal & placental risk factors on low birth weight neonates

A Prospective study on the effects & prevalence of maternal, fetal & placental risk factors on low birth weight neonates

Placental Factors Affecting Birth Weight of Late Onset Severe Preeclampsia

Objective of the study is to determine the correlation of placenta and birth weight in late onset preeclampsia. A case-control study,analysis prevalence was conducted in Moh Hoesin Hospital Palembang from August 2015 to August 2016. Samples were women who giving birth in Moh Hoesin Hospital Palembang. They were devided into two groups, severe preeclampsia as case group and normotension as control. Data were analyzed by X2, Exact Fisher’s and logistic regression test using SPSS 16.0. There were 180 subjects (90 cases and 90 controls). There was a positive correlation between placental macroscopic and late onset preeclampsia (p=0.009; OR=6.9), in contrast there was only one different placental microvascularisation of 16, between late onset preeclampsia and normotension, the mural or occlusive fibrin thrombi chorion (p=0.005; OR=9.9). Birth weight in late onset preeclampsia tended to be small but still in normal range and it was not statistictly significant (p=0.112; OR=10.4). There was a positive correlation between placental macroscopic and SGA baby on late onset preeclampsia (p=0.026; OR=16.6), but it wasnot proven microscopically. Placenta remains contributed to the pathogenesis of the late onset preeclampsia, but not as dominant as the early one.

Placental Factors Affecting Birth Weight of Late Onset Severe Preeclampsia

Preeclampsia (PE) is one of the leading causes of maternal and perinatal mortality and morbidity. Placental insufficiency is considered as the main pathogenesis in the early onset PE. Objective of the study is to determine the correlation of placenta and birth weight in late onset preeclampsia. A case-control study of prevalence was conducted in Moh. Hoesin Hospital Palembang from August 2015 to August 2016. Samples were women who giving birth in Moh.Hoesin Hospital Palembang. They were divided into two groups, severe preeclampsia as case group and normotensionas control. Data were analyzed by X2, Exact Fisher’s and logistic regression test using SPSS 16.0. There were 180 subjects (90 cases and 90 controls). There was a positive correlation between placental macroscopic and late onset preeclampsia (p=0.009; OR=6.9), in contrast there was only one different placental microvascularisation of 16, between late onset preeclampsia and normotension, the mural or occlusive fibrin thrombi chorion (p=0.005; OR=9.9). Birth weight in late onset preeclampsia tended to be small but still in normal range and it was not statistictly significant (p=0.112; OR=10.4). There was a positive correlation between placental macroscopic and SGA baby on late onset preeclampsia (p=0.026; OR=16.6), but it wasnot proven microscopically. Placenta remains contributed to the pathogenesis of the late onset preeclampsia, but not as dominant as the early one.

Reduced fetal movement intervention Trial-2 (ReMIT-2): protocol for a pilot randomised controlled trial of standard care informed by the result of a placental growth factor (PlGF) blood test versus standard care alone in women presenting with reduced fetal movement at or after 36+\u20090 weeks gestation

BackgroundForty percent of babies who are stillborn born die after 36 weeks gestation and have no lethal structural abnormality. Maternal perception of reduced fetal movement (RFM) is associated with stillbirth and is related to abnormal placental structure and function. The ultimate objective of this trial is to assess whether for women with RFM, intervention directed by measurement of placental biochemical factors in addition to standard care improves pregnancy outcome compared with standard care alone. This is the protocol for a pilot trial to determine the feasibility of a definitive trial and also provide proof of concept that informing care by measurement of placental factors improves neonatal outcomes.MethodsReMIT-2 is a multicentre, pilot randomised controlled trial of care informed by results of an additional placental factor blood test versus standard care alone for women presenting with RFM at or after 36+ 0 weeks gestation. Participants will be randomised 1:1 to the intervention arm where the blood test result is revealed and acted on, or to the control arm where the blood sample is not tested immediately and therefore the result cannot be acted on. All participants will be followed up six weeks after delivery to assess their health status and views of the trial, along with healthcare costs. A sub-group will be interviewed within 16 weeks after delivery to further explore their views of the trial. Outcomes to determine feasibility of a definitive trial include number of potentially eligible women, proportion lost to follow-up, clinical characteristics at randomisation, reasons for non-recruitment, compliance with the trial intervention and views of participants and clinicians about the trial. Proof of concept outcomes include: rates of induction of labour; Caesarean birth; and a composite neonatal outcome of stillbirths and deaths before discharge, 5-min Apgar score < 7, umbilical artery pH < 7.05 and admission to neonatal unit for > 48 h.DiscussionResults from this pilot trial will help determine whether a large definitive trial is feasible. Such a study would provide evidence to guide management of women with RFM and reduce stillbirths.Trial registrationISRCTN Registry, ISRCTN12067514. Registered on 8 September 2017.

OP21.07: Correlation between placental pathology and angiogenetic factors

OP21.07: Correlation between placental pathology and angiogenetic factors

89. Placental oxidative stress, angiogenic factor and maternal endothelial function in pregnant women with normotensive fetal growth restriction

Introduction Placental oxidative damage may increase the production of anti-angiogenic factors, resulting in maternal endothelial dysfunction in pregnant women with preeclampsia (PE). Endothelial dysfunction is observed in women with a history of normotensive fetal growth restriction (FGR). However, placental oxidative stress, angiogenic factors and maternal endothelial function in those women remain unclear. Objectives This study aimed to assess the relationship between placental oxidative stress, angiogenic factors and maternal endothelial function in pregnant women with normotensive FGR. Methods A total of 21 women with uncomplicated pregnancies, 17 women with early-onset preeclampsia (PE), 18 with late-onset PE, and 14 with normotensive FGR were evaluated. We measured to determine serum parameters of oxygen free radicals (d-ROMs), PlGF and sFlt-1 as maternal anti-angiogenic factor, placental oxidative DNA damage, and flow-mediated vasodilation (FMD) as a marker of maternal endothelial function. Immunohistochemical analysis was performed to measure the proportion of placental trophoblast cell nuclei staining positive for 8-hydroxy-2′-deoxyguanosine (8-OHdG) and redox factor-1(ref-1), which are markers of oxidative DNA damage. Results Maternal serum d-ROMs, sFlt-1 concentrations, and FMD did not significantly differ between control and normotensive FGR groups. The proportion of nuclei staining positive for 8-OHdG was significantly higher in the normotensive FGR group relative to the control group, but ref-1 were not significantly differences among all groups. Conclusion Our findings demonstrate that although placental oxidative DNA damage was observed in both women with PE and those with normotensive FGR, pregnant women with normotensive FGR show no increase in the concentrations of sFlt-1 and d-ROMs, or a decrease in FMD.

35. Early- and late-onset of pre-eclampsia: How do the maternal circulating biomarkers differ, and why?

Pre-eclampsia requires the presence of placenta or residual placental compounds (as in postpartum pre-eclampsia), but the relative contribution of maternal predisposing factors versus placental factors to its pathophysiology is not well understood. The talk will revisit the classical “two-stage model” of pre-eclampsia, as proposed by Redman et al. in 1999, where incomplete placentation is the first of two stages of pre-eclampsia, typically of early onset. Maternal “intolerance” of fetal cells and failed uteroplacental spiral artery remodelling has been a proposed pathway for poor placentation, which however involves several mechanisms. The second stage comprises dysfunctional uteroplacental perfusion and placental oxidative stress, followed by secretion of inflammatory factors to the maternal circulation, with ensuing generalized maternal vascular inflammation and pre-eclampsia signs (maternal hypertension and proteinuria). We have suggested (Redman, Sargent and Staff, Placenta 2013) that the mechanisms leading to a dysfunctional uteroplacental circulation may involve additional mechanisms to poor placentation, and that syncytiotrophoblast (STB) stress, with upregulation of some proteins (as exemplified by “antioangiogenic proteins”) and downregulation of others (exemplified by “proangiogenic proteins”) is the common final pathway. We argue that maternal circulating angiogenic factors are markers of STB stress, not of preeclampsiaper se. We have proposed two major forms of placental dysfunction in pre-eclampsia: one extrinsic (poor placentation) and one intrinsic cause (villous overcrowding). Our model explains important differences of early- and late-onset pre-eclampsia, including their relation to maternal circulating placenta-associated proteins used as biomarkers for the syndrome (Redman and Staff, AJOG 2015). The talk will also argue that there is no condition such as “maternal” pre-eclampsia, as all pre-eclampsia originate in the placenta. We suggest however that maternal factors may contribute two both stages of pre-eclampsia, the first by affecting either placental pathways to STB stress and the latter by amplifying the effects of STB stress on maternal vasculature.

45. Effects of syncytiotrophoblast extracellular vesicles (STBEVs) on endothelium-dependent vasodilation in uterine arteries from lectin-like oxidized ldl receptor-1 (LOX-1) overexpressing mice during pregnancy

Introduction Placental factors, such as syncytiotrophoblast extracellular vesicles (STBEVs), have been suggested to contribute to maternal endothelial vascular dysfunction in women with preeclampsia (PE). The lectin-like oxidized LDL receptor-1 (LOX-1) is a multi-ligand scavenger receptor; and in women with PE both STBEV levels and LOX-1 expression are elevated. We used heterozygous LOX-1 overexpressing (LOX-1OE) mice to further investigate the role of LOX-1 and STBEVs in vascular dysfunction during pregnancy. Hypothesis We hypothesized that STBEVs activate LOX-1 and contribute to vascular dysfunction in mouse uterine arteries of LOX-1OE mice, but not in WT controls. Methods Uterine arteries were obtained from late pregnant (gestational day 18; term = day 19) LOX-1OE mice (carrying a bovine LOX-1 transgene) and WT controls (−/−). Isolated vessels were incubated overnight in the absence (n = 8) or presence of STBEVs (200 μg/ml, n = 5–7) from control pregnant women. Using wire myography, endothelium-dependent vasodilation responses to methylcholine (MCh) were assessed with or without L-NAME (pan nitric oxide synthase inhibitor), superoxide dismutase (SOD) or oxidized LDL (oxLDL). Results Endothelium-dependent vasodilation to MCh was unchanged following STBEV-incubation in both mouse strains. In WT mice, nitric oxide did not contribute to vasodilation, while there was a nitric oxide contribution to vasodilation in STBEV-incubated arteries (p Discussion Interestingly, both LOX-1OE and STBEV-exposure increased nitric oxide contribution to relaxation; perhaps as a compensatory mechanism. OxLDL (a potential contributing factor in women with PE and dyslipidemia) impaired endothelial-dependent relaxation in the presence of increased LOX-1 expression during pregnancy. These data suggest that increased LOX-1 expression in women with PE could contribute to impaired vascular function.

Villous Capillary Lesions OfPlacentaA Ten-Year Experience With Brief Review Of Literature

Background: Villous capillary (VC) lesions of placenta range from reactive to benign tumors such as chorangiosis (CH), chorangiomatosis (CM) and chorangioma (CA). Associated with perinatal morbidity and mortality, these rare lesions are documented with maternal, placental and fetal risk factors. The aim of this study was to analyze the clinico-pathological profile of VC lesions of placenta and to compare the associated risk factors between VC lesion (CH) and those with no VC lesions. Methods: This retrospective study includes all the VC lesions of Placenta diagnosed in Pondicherry Institute of medical sciences from January 2006 to February 2016. For comparison, gestational age matched controls with no VC lesions were obtained. Chi-square test or fischer’s exact test was used for statistical analysis. Results: Of 29 VC lesions of placenta (5.6%) in 10 years, 27 were CH. Commonly associated feto-maternal factors include anemia (47.4%), oligohydramnia (33.3%), infection (28.6%), pre-eclampsia (23.8%), NICU admission (47.8%), fetal growth retardation (17.4%), congenital anomaly (17.4%), intrauterine death (13%). When compared with controls, CH had significantly increased LSCS, premature rupture of membranes and histologically, chorioamnionitis and funisitis that support its infective etiology. One case of CA associated with IUGR, polyhydramnios was found to have CH in her next pregnancy which points to a possibility of genetic predisposition among VC lesions. CM was diagnosed in primi with severe pre-eclampsia and asymmetric IUGR. Conclusion: Prevalence of CH is 5.2%. VC lesions of placenta are commonly associated with complicated pregnancy and neonatal morbidity.

The role of placental malperfusion in the pathogenesis of preeclampsia in dichorionic twin and singleton pregnancies

IntroductionIn singletons, the pathogenesis of hypertensive disorders of pregnancy (HDP) is attributed to abnormal placentation, characterized by maternal vascular malperfusion (MVM) lesions. Whether MVM plays a similar role in twin pregnancies is unclear. The purpose of the study was to compared placental pathology findings between dichorionic-twin and singleton pregnancies complicated by HDP. MethodsRetrospective cohort study of women with dichorionic-twin or singleton pregnancies complicated by HDP who gave birth in a single tertiary center between 2001 and 2015. Placental abnormalities were classified into lesions associated with MVM, fetal vascular malperfusion, placental hemorrhage and chronic villitis. Placental findings and neonatal outcomes were compared between twin and singleton pregnancies. ResultsA total of 144 women with twins and 768 women with a singleton pregnancy met the inclusion criteria. Compared with HDP singletons, twins with HDP had higher mean birth weights, were less likely to be small for gestational age and be born at <34 and at <32 weeks. Twins had lower odds for placental weight below <10th percentile (aOR 0.49, 95%CI 0.33–0.71), for MVM pathology (aOR 0.28, 95%CI 0.20–0.39) and for fetal vascular malperfusion pathology (aOR 0.65, 95%CI 0.45–0.93). These finding remained significant in the subpopulation of early onset HDP (<34 weeks) and small for gestational newborn. DiscussionOur findings support the hypothesis that MVM are less relevant to the pathogenesis of HDP in twin pregnancies and suggest that other placental or non-placental factors are responsible for the increased risk of HDP in twin pregnancies.

HELLP-синдром: клинико-лабораторные особенности и дисбаланс плацентарных факторов ангиогенеза

HELLP-синдром: клинико-лабораторные особенности и дисбаланс плацентарных факторов ангиогенеза

Upregulation of Angiogenic Factors via Protein Kinase C and Hypoxia-induced Factor-1α Pathways under High-glucose Conditions in the Placenta.

Abnormal glucose metabolism during pregnancy is an established risk factor for preeclampsia (PE). Disruption of the balance between placental angiogenic factors is linked to PE pathophysiology. We examined whether hypoxia-induced factor-1α (HIF-1α) and protein kinase Cβ (PKCβ) are involved in the regulation of placental angiogenic factors under high-glucose conditions in vitro. The human choriocarcinoma cell lines BeWo and JEG-3, and the human trophoblast cell line HTR-8/SVneo were cultured with 10 and 25 mmol/L glucose [control glucose group (CG) and high-glucose group (HG), respectively]. We examined the changes in HIF-1α, soluble fms-like tyrosine kinase-1 (sFlt-1), placental growth factor (PlGF), and vascular endothelial growth factor (VEGF) expression in the CG and HG by real-time PCR and ELISA. PKC activation was also measured by ELISA. The expressions of HIF-1α, sFlt-1, PlGF, and VEGF were significantly higher in the HG than in the CG. PKC activity was significantly increased in the HG. High glucose affected the expression of angiogenic factors in choriocarcinoma cells via the PKCβ and HIF-1α pathways, suggesting their involvement in PE pathogenesis.

Physiological Changes in Cardiovascular System during Normal Pregnancy: A Review

AbstractPregnancy is a complex biological process associated with changes in physiologic functions of the body. Dramatic changes take place in the cardiovascular physiology leading to gradual adaptation of these changes by the body of the pregnant woman. Cardiac output increases during pregnancy to 30 to 50% above the prepregnant levels. The increase in cardiac output occurs due to increase in stroke volume initially during gestation and later by increase in heart rate. These changes in cardiac output are attributed to either neurohumoral factors such as estrogen and progesterone or placental factors. Maternal body position affects cardiac output with highest in kneel-chest and left lateral positions. Along with these changes, variations in heart rate, blood pressure, and blood volume are observed following a specific pattern of change during pregnancy. Hence, it is necessary to understand the cardiovascular changes during pregnancy to interpret, predict, and diagnose any cardiac disease efficiently.

Плацентарний синдром як імовірний фактор ризику розвитку прееклампсії (Огляд літератури)

The modern studies review summarizes the data of recent years, which covers issues related to the preeclampsia pathogenesis study on the placental disturbances theory basis. The model of placental preeclampsia and factors that change before the development of its clinical manifestations a considered. Attention is paid to the placental factors possible combination analysis the are important in preventing the disease in terms of prognostic value. Key words: preeclampsia, placental dysfunction, biochemical markers, placental weight.

Impact of congenital cytomegalovirus infection on transcriptomes from archived dried blood spots in relation to long-term clinical outcome.

Congenital Cytomegalovirus infection (cCMV) is the leading infection in determining permanent long-term impairments (LTI), and its pathogenesis is largely unknown due to the complex interplay between viral, maternal, placental, and child factors. The cellular activity, considered to be the result of the response to exogenous and endogenous factors, is captured by the determination of gene expression profiles. In this study, we determined whole blood transcriptomes in relation to cCMV, CMV viral load and LTI development at 6 years of age by using RNA isolated from neonatal dried blood spots (DBS) stored at room temperature for 8 years. As DBS were assumed to mainly reflect the neonatal immune system, particular attention was given to the immune pathways using the global test. Additionally, differential expression of individual genes was performed using the voom/limma function packages. We demonstrated feasibility of RNA sequencing from archived neonatal DBS of children with cCMV, and non-infected controls, in relation to LTI and CMV viral load. Despite the lack of statistical power to detect individual genes differences, pathway analysis suggested the involvement of innate immune response with higher CMV viral loads, and of anti-inflammatory markers in infected children that did not develop LTI. Finally, the T cell exhaustion observed in infected neonates, in particular with higher viral load, did not correlate with LTI, therefore other mechanisms are likely to be involved in the long-term immune dysfunction. Despite these data demonstrate limitation in determining prognostic markers for LTI by means of transcriptome analysis, this exploratory study represents a first step in unraveling the pathogenesis of cCMV, and the aforementioned pathways certainly merit further evaluation.