Kliinilise auditi „Iseseisva antenataalse ämmaemandusabi kvaliteet“ kokkuvõte

Eesti Arst 2017; 96(7):386–38

Extensive shift in placental transcriptome profile in preeclampsia and placental origin of adverse pregnancy outcomes

One in five pregnant women suffer from gestational complications, prevalently driven by placental malfunction. Using RNASeq, we analyzed differential placental gene expression in cases of normal gestation, late-onset preeclampsia (LO-PE), gestational diabetes (GD) and pregnancies ending with the birth of small-for-gestational-age (SGA) or large-for-gestational-age (LGA) newborns (n = 8/group). In all groups, the highest expression was detected for small noncoding RNAs and genes specifically implicated in placental function and hormonal regulation. The transcriptome of LO-PE placentas was clearly distinct, showing statistically significant (after FDR) expressional disturbances for hundreds of genes. Taqman RT-qPCR validation of 45 genes in an extended sample (n = 24/group) provided concordant results. A limited number of transcription factors including LRF, SP1 and AP2 were identified as possible drivers of these changes. Notable differences were detected in differential expression signatures of LO-PE subtypes defined by the presence or absence of intrauterine growth restriction (IUGR). LO-PE with IUGR showed higher correlation with SGA and LO-PE without IUGR with LGA placentas. Whereas changes in placental transcriptome in SGA, LGA and GD cases were less prominent, the overall profiles of expressional disturbances overlapped among pregnancy complications providing support to shared placental responses. The dataset represent a rich catalogue for potential biomarkers and therapeutic targets.Peer reviewe

Hõbekuulid vähiteraapias: teel suunatud vähiravi poole

Vähiuuringute üheks keskseks eesmärgiks on vähirakke normaalsetest rakkudest eristavate ravimite väljatöötamine. Kliinilises kasutuses olevate vähiravimite puuduseks on nende vähene selektiivsus ja sellest tingitud kõrvalmõjud normaalsetes kudedes. Artiklis on antud ülevaade suunatud vähiravimite valjatöötamisest ja suundumustest. Suunatud vähiravi põhineb molekulaarsetel erinevustel vähikoe ja normaalsete veresoonte vahel. In vivo faagidisplei meetodi abil on võimalik veresoonte haigusspetsiifilisi molekulaarseid mustreid kaardistada. Selle tulemuseks on peptiidid, mis seostuvad selektiivselt vähiveresoontega. Need kullerpeptiidid nagu ka teised kasvaja veresoontega selektiivselt seonduvad molekulid (antikehad, aptameerid) võimaldavad kasvajasse viia vähiravimeid ja kontrastaineid. Hiljuti avastatud kasvajakude penetreerivad peptiidid põhjustavad vähiveresoonte selektiivset lekkimist. Koos ravimitega manustatuna põhjustavad need peptiidid ravimi väljumist kasvaja veresoontest ja tungimist kasvajakoesse – tulemuseks on ravimi terapeutilise indeksi paranemine.Eesti Arst 2015; 94(5):281–28

Mid-gestational gene expression profile in placenta and link to pregnancy complications.

Despite the importance of placenta in mediating rapid physiological changes in pregnancy, data on temporal dynamics of placental gene expression are limited. We completed the first transcriptome profiling of human placental gene expression dynamics (GeneChips, Affymetrix®; ~47,000 transcripts) from early to mid-gestation (n = 10; gestational weeks 5-18) and report 154 genes with significant transcriptional changes (ANOVA, FDR P<0.1). TaqMan RT-qPCR analysis (n = 43; gestational weeks 5-41) confirmed a significant (ANOVA and t-test, FDR P<0.05) mid-gestational peak of placental gene expression for BMP5, CCNG2, CDH11, FST, GATM, GPR183, ITGBL1, PLAGL1, SLC16A10 and STC1, followed by sharp decrease in mRNA levels at term (t-test, FDR P<0.05). We hypothesized that normal course of late pregnancy may be affected when genes characteristic to mid-gestation placenta remain highly expressed until term, and analyzed their expression in term placentas from normal and complicated pregnancies [preeclampsia (PE), n = 12; gestational diabetes mellitus (GDM), n = 12; small- and large-for-gestational-age newborns (SGA, LGA), n = 12+12]. STC1 (stanniocalcin 1) exhibited increased mRNA levels in all studied complications, with the most significant effect in PE- and SGA-groups (t-test, FDR P<0.05). In post-partum maternal plasma, the highest STC1 hormone levels (ELISA, n = 129) were found in women who had developed PE and delivered a SGA newborn (median 731 vs 418 pg/ml in controls; ANCOVA, P = 0.00048). Significantly higher expression (t-test, FDR P<0.05) of CCNG2 and LYPD6 accompanied with enhanced immunostaining of the protein was detected in placental sections of PE and GDM cases (n = 15). Our study demonstrates the importance of temporal dynamics of placental transcriptional regulation across three trimesters of gestation. Interestingly, many genes with high expression in mid-gestation placenta have also been implicated in adult complex disease, promoting the discussion on the role of placenta in developmental programming. The discovery of elevated maternal plasma STC1 in pregnancy complications warrants further investigations of its potential as a biomarker

The immunostaining of CCNG2 and LYPD6 proteins was assessed in placental sections from term pregnancies with no complications (controls), with preeclampsia (PE) or with gestational diabetes mellitus (GDM).

<p>(A) Hematoxylin-eosin staining was used to describe histopathological findings in analyzed placental samples (100-fold microscope magnification). In term placentae the mature intermediate (IMV) and small terminal villi (STV) were seen. Characteristic to PE, villous agglutination and infarction (IN; intense eosinophilic staining) and increased number of syncytial knots (SK) were detected. GDM presented with degenerative placental lesions such as focal villous fibrinoid necrosis (FN). (B) Diffuse cytoplasmic staining of LYPD6 antibody was detected in syncytiotrophoblast (ST) cells in all villous types (IMV, STV). Additionally LYPD6 antibody strongly stained the cytoplasm and the nucleus of villous stroma Hoffbauer cells (H), fibroblasts (F) and endothelial cells (E) of villous vessels. No localization differences in LYPD6 antibody stain between the groups were found; however strong tendency to higher staining intensity was observed in PE and GDM placentas compared to normal term placenta. (C) CCNG2 antibody showed fine granular cytoplasmic staining of villous stromal Hoffbauer (H) and fibroblast (F) cells. In addition, weak cytoplasmic staining of syncytiotrophoblast (ST) and endothelial cells (E) of vessel wall was found. No localization differences in CCNG2 staining between the normal, PE and GDM groups were detected. Higher tendency to positivity was seen in PE placental sections. (D) Negative control (NC) staining was performed without primary antibody. Scale bar, 100 µm. Microscope magnifications ×100 and ×400 were used. Brown color indicates chromogen-labeled antibody and blue color indicates hematoxylin nuclear staining.</p

Genes with significant dynamic increase in expression during early and mid-gestation as experimentally confirmed by RT-qPCR.

<p>Relative mRNA expression levels in the extended sample set of first and second trimester placentas (<i>n</i> = 31; from gestational days 38 to 147) were determined by TaqMan assays. <i>P-</i>values were calculated by ANOVA and subjected to multiple testing correction (FDR). Genes below the significance threshold of <i>P</i>-value>0.02 are shown in <b><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0049248#pone.0049248.s007" target="_blank">Figure S4</a></b>.</p

Molecular functions of identified mid-gestation marker genes in placenta and their involvement in clinical conditions.

a<p>References are listed in <b><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0049248#pone.0049248.s018" target="_blank">Table S9</a></b>.</p

Expressional dynamics of identified mid-pregnancy specific genes in placenta from early to term gestation.

<p>Relative mRNA levels were determined by RT-qPCR TaqMan assays in placental tissues from early- (5–13 gestational weeks; <i>n</i> = 23), mid- (17–21 gestational weeks; <i>n</i> = 8) and term-gestation (36–41 gestational weeks; <i>n</i> = 12) samples of uncomplicated pregnancy cases. Boxplots show mid-gestation marker genes with (A) significantly increased mRNA expression compared to early- or late-gestation placental samples, (B) significantly increased expression levels compared to early gestation placental samples, and (C) gradual increase in expression during pregnancy. <i>P</i>-values were calculated by Student t-test and subjected to multiple testing correction (FDR) (<b><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0049248#pone.0049248.s016" target="_blank">Table S7</a></b>).</p

Maternal and offspring characteristics of REPROMETA samples used in the study for RT-qPCR and ELISA experiments.

<p>Data are given as medians with ranges, except where indicated differently.</p><p>Nulliparity  =  no previous childbirth.</p>*<p><i>P</i><0.05 <i>vs</i>. control group, Mann-Whitney U or Fisher’s exact test.</p><p>SGA, small-for-gestational age; LGA, large-for-gestational age; PE, preeclampsia; GDM, gestational diabetes mellitus; yr, years; d, days.</p