Dysregulation of micro-RNA contributes to the risk of unexplained recurrent pregnancy loss

Although there are plenty of evidence that dysregulation of microRNA (miRNA) level is involved in many human diseases, it is still unknown whether abnormal levels of specific miRNAs are associated with recurrent pregnancy loss (RPL). We believe that such an association do exist as this study confirmed significant differences in the level of specific miRNAs between RPL cases and healthy controls. The study was conducted on 100 Palestinian women: 60 patients with at least two unexplained consecutive pregnancy losses half of them were pregnant at the first trimester and the rest were non-pregnant and 40 healthy controls with at least two live births and no history of pregnancy loss; half of them were at their first trimester of pregnancy and the rest were non-pregnant. We investigated the relative expression of miR-21, miR-126, miR-155, miR-182, miR-222 and miR-517* using quantitative real-time polymerase chain reaction and Ct method experiments. Differential expression was evaluated using Student t-test and fold change analyses. The expression difference of miR-21, miR-126 and miR-182 between patients and controls in the pregnant subjects showed statistically significant difference (p-value ≤ 0.05) with fold decrease of 1.5, 1.6 and 5.6, respectively. In the non-pregnant group miR-21, miR-126, miR-222 and miR-517* expressions were significantly different with fold decrease of 2.4, 2.9, 2.7 and 11.8, respectively. In conclusion, at least miR-21 and miR-126 could serve as potential markers for idiopathic RPL as their levels were significantly lower in patients before being pregnant and during pregnancy. Moreover, restoration of the normal level of those miRNAs might be a novel intervention strategy in unexplained RPL

Role of IL-36 Cytokines in the Regulation of Angiogenesis Potential of Trophoblast Cells

IL-36 cytokines (the agonists IL-36α, IL-36β, IL-36γ, and the antagonist IL-36Ra) are expressed in the mouse uterus and associated with maternal immune response during pregnancy. Here, we characterize the expression of IL-36 members in human primary trophoblast cells (PTC) and trophoblastic cell lines (HTR-8/SVneo and JEG-3) and upon treatment with bacterial and viral components. Effects of recombinant IL-36 on the migration capacity of trophoblastic cells, their ability to interact with endothelial cells and the induction of angiogenic factors and miRNAs (angiomiRNAs) were examined. Constitutive protein expression of IL-36 (α, β, and γ) and their receptor (IL-36R) was found in all cell types. In PTC, transcripts for all IL-36 subtypes were found, whereas in trophoblastic cell lines only for IL36G and IL36RN. A synthetic analog of double-stranded RNA (poly I:C) and lipopolysaccharide (LPS) induced the expression of IL-36 members in a cell-specific and time-dependent manner. In HTR-8/SVneo cells, IL-36 cytokines increased cell migration and their capacity to interact with endothelial cells. VEGFA and PGF mRNA and protein, as well as the angiomiRNAs miR-146a-3p and miR-141-5p were upregulated as IL-36 response in PTC and HTR-8/SVneo cells. In conclusion, IL-36 cytokines are modulated by microbial components and regulate trophoblast migration and interaction with endothelial cells. Therefore, a fundamental role of these cytokines in the placentation process and in response to infections may be expected

Association between Selected Maternal Plasma Micro-RNAs and Idiopathic Recurrent Pregnancy Loss

MicroRNAs are small noncoding RNAs that function to control gene expression. Recurrent pregnancy loss (RPL) is two or more consecutive pregnancy losses before 20 weeks of gestation. The aim of this study is to explore the expression level of a group of pregnancy-associated miRNAs in maternal plasma in normal pregnancy and RPL cases. We conducted a case control study on 100 Palestinian women: 60 patients with at least two unexplained consecutive pregnancy losses half of them were pregnant at the first trimester and the rest were non-pregnant and 40 healthy controls with at least two live births and no history of pregnancy loss; half of them were at their first trimester of pregnancy and the rest were non-pregnant. We investigated the relative expression of miR-21, miR-126, miR-155, miR-182, miR-222 and miR-517* using quantitative real-time polymerase chain reaction and Ct method experiments. Differential expression was evaluated using Student t test and fold change analyses. Only the expression difference of miR-21, miR-126 and miR-182 between patients and controls in the pregnant group showed statistically significant difference (p-value ≤ 0.05 ) with fold decrease of 1.5, .1.6 and 5.6, respectively. In non-pregnant group miR-21, miR-126, miR-222 and miR-517* expression were significantly different with fold decrease of 2.4, 2.9, 2.7 and 11.8, respectively. So miR-21 and miR-126 proved to be the most important microRNAs in idiopathic RPL as their level was significantly decreased in patients before being pregnant and during pregnancy

MicroRNA in a case of unexplained recurrent pregnancy loss

The present report describes the relative quantitation of a group of selected miRNAs in a twenty one years old Palestinian woman who experienced 8 unexplained first trimester pregnancy losses. The relative expression of miR-21, miR-126, miR-155, miR-182, miR-222 and miR-517* in her plasma was measured before pregnancy and in the seventh week of her ninth pregnancy. The relative expression of the same miRNAs was also measured before pregnancy and in the eighth week of fifth pregnancy in a healthy control woman who has four live children and without any history of pregnancy loss. The results of the case woman showed an extremely decreased level of all the investigated microRNAs during her pregnancy as compared to their level before being pregnant, except for miR-517* which was elevated. The relative quantitation of miR-21, miR-126, miR-155, miR-182, miR-222 and miR-517* was 0.026, 0.12, 0.23, 0.26, 0.28 and 1.4, respectively. However, the relative quantitation of the respective miRNAs in the control was 0.92, 0.75, 0.6, 1.1, 1.11 and 10.9. Unfortunately, at the end of eighth week of her pregnancy the case woman lost her fetus. We believe that the dysregulation of miRNA is involved in the recurrent pregnancy losses in the case presented in this report

Role of IL-36 Cytokines in the Regulation of Angiogenesis Potential of Trophoblast Cells

IL-36 cytokines (the agonists IL-36α, IL-36β, IL-36γ, and the antagonist IL-36Ra) are expressed in the mouse uterus and associated with maternal immune response during pregnancy. Here, we characterize the expression of IL-36 members in human primary trophoblast cells (PTC) and trophoblastic cell lines (HTR-8/SVneo and JEG-3) and upon treatment with bacterial and viral components. Effects of recombinant IL-36 on the migration capacity of trophoblastic cells, their ability to interact with endothelial cells and the induction of angiogenic factors and miRNAs (angiomiRNAs) were examined. Constitutive protein expression of IL-36 (α, β, and γ) and their receptor (IL-36R) was found in all cell types. In PTC, transcripts for all IL-36 subtypes were found, whereas in trophoblastic cell lines only for IL36G and IL36RN. A synthetic analog of double-stranded RNA (poly I:C) and lipopolysaccharide (LPS) induced the expression of IL-36 members in a cell-specific and time-dependent manner. In HTR-8/SVneo cells, IL-36 cytokines increased cell migration and their capacity to interact with endothelial cells. VEGFA and PGF mRNA and protein, as well as the angiomiRNAs miR-146a-3p and miR-141-5p were upregulated as IL-36 response in PTC and HTR-8/SVneo cells. In conclusion, IL-36 cytokines are modulated by microbial components and regulate trophoblast migration and interaction with endothelial cells. Therefore, a fundamental role of these cytokines in the placentation process and in response to infections may be expected