Project description:Abstract Objective: To investigate the physiology of ectopic pregnancy by analyzing the miRNA profiles of the pregnancy tissue from both ectopic and control pregnancies (voluntary termination of pregnancy). Design: Research study. Setting: Academic research institute in collaboration with a university hospital. Patients: Patients suffering from tubal EP and patients with a normal ongoing pregnancy scheduled for a voluntary termination of pregnancy (VTOP) were recruited. Interventions: Pregnancy tissue samples were analyzed by miRNA microarray and further validated by qPCR. Main Outcome measures: Gene expression profiles and quantitative PCR measurements. Results: Four miRNAs were found to be downregulated in EP compared to healthy trophoblast tissue, and three miRNAs were upregulated in EP trophoblast compared to control tissue samples. All genes were validated by q-PCR. We found three statistically significant miRNAs, all of which were upregulated in the EP sample group. Conclusion: We describe the alteration of seven miRNAs in EP samples that could alter pathways which are critical for correct implantation such as extracellular matrix (ECM) remodeling, or mucin biosynthesis, thus resulting in ectopic pregnancies.
Project description:Abstract Objective: To investigate the physiology of ectopic pregnancy by analyzing the miRNA profiles of the pregnancy tissue from both ectopic and control pregnancies (voluntary termination of pregnancy). Design: Research study. Setting: Academic research institute in collaboration with a university hospital. Patients: Patients suffering from tubal EP and patients with a normal ongoing pregnancy scheduled for a voluntary termination of pregnancy (VTOP) were recruited. Interventions: Pregnancy tissue samples were analyzed by miRNA microarray and further validated by qPCR. Main Outcome measures: Gene expression profiles and quantitative PCR measurements. Results: Four miRNAs were found to be downregulated in EP compared to healthy trophoblast tissue, and three miRNAs were upregulated in EP trophoblast compared to control tissue samples. All genes were validated by q-PCR. We found three statistically significant miRNAs, all of which were upregulated in the EP sample group. Conclusion: We describe the alteration of seven miRNAs in EP samples that could alter pathways which are critical for correct implantation such as extracellular matrix (ECM) remodeling, or mucin biosynthesis, thus resulting in ectopic pregnancies. This study was authorized by the Institutional Review Board/Independent Ethics Committee of the Hospital Universitario La Fe, Valencia, Spain. Eight patients suffering from tubal EP and another eight patients with a normal ongoing pregnancy scheduled for a VTOP were recruited.
Project description:We have sequenced miRNA libraries from human embryonic, neural and foetal mesenchymal stem cells. We report that the majority of miRNA genes encode mature isomers that vary in size by one or more bases at the 3’ and/or 5’ end of the miRNA. Northern blotting for individual miRNAs showed that the proportions of isomiRs expressed by a single miRNA gene often differ between cell and tissue types. IsomiRs were readily co-immunoprecipitated with Argonaute proteins in vivo and were active in luciferase assays, indicating that they are functional. Bioinformatics analysis predicts substantial differences in targeting between miRNAs with minor 5’ differences and in support of this we report that a 5’ isomiR-9-1 gained the ability to inhibit the expression of DNMT3B and NCAM2 but lost the ability to inhibit CDH1 in vitro. This result was confirmed by the use of isomiR-specific sponges. Our analysis of the miRGator database indicates that a small percentage of human miRNA genes express isomiRs as the dominant transcript in certain cell types and analysis of miRBase shows that 5’ isomiRs have replaced canonical miRNAs many times during evolution. This strongly indicates that isomiRs are of functional importance and have contributed to the evolution of miRNA genes
Project description:Ectopic pregnancy is the leading cause of morbidity and mortality among women in the first trimester. A reliable diagnostic test for the detection of ectopic pregnancy in cases of non-viable pregnancy of unknown location (NV-PUL) is needed. The objective of the current study is to prospectively validate our previous findings of a 12-cilia gene classifier for the identification of ectopic pregnancy. Molecular interrogation is performed to define differences in molecular expression profiles in ectopic and abnormal intrauterine pregnancies. These samples come from a prospective cohort study comparing transcriptome profiles from women with ectopic pregnancy (ECT) or non-viable intrauterine pregnancy (NV-IUP). Samples were analyzed with the Affymetrix Human Gene 2.0 ST Array.
Project description:To characterize the human plasma microtranscriptome profile at first trimester of pregnancy in presence or not of pregnancy complications, we sequenced microRNAs in plasma samples collected from pregnant women between the 6th and the 15th weeks of pregnancy as a replication cohort. We then performed differential expression analyses to assess the miRNA profile diffrences according to the presence of pregnancy complications or not (i.e. Gestational diabetes mellitus, Gestational hypertension or preeclampsia vs. normal pregnancies).
Project description:Ectopic pregnancy is the leading cause of morbidity and mortality among women in the first trimester. A reliable diagnostic test for the detection of ectopic pregnancy in cases of non-viable pregnancy of unknown location (NV-PUL) is needed. The objective of the current study is to define differences in molecular expression profiles in ectopic and abnormal intrauterine pregnancies toward discovery of biologically plausible ectopic classifier candidate. These samples come from a prospective cohort study comparing transcriptome profiles from women with ectopic pregnancy (ECT) or non-viable intrauterine pregnancy (NV-IUP). Samples were analyzed with the Affymetrix Human Gene 2.0 ST Array. The views expressed are those of the author(s) and do not reflect the official policy of the Department of the Army, the Department of Defense or the U.S. Government. The investigators have adhered to the policies for protection of human subjects as prescribed in 45 CFR 46.
Project description:We have sequenced miRNA libraries from human embryonic, neural and foetal mesenchymal stem cells. We report that the majority of miRNA genes encode mature isomers that vary in size by one or more bases at the 3’ and/or 5’ end of the miRNA. Northern blotting for individual miRNAs showed that the proportions of isomiRs expressed by a single miRNA gene often differ between cell and tissue types. IsomiRs were readily co-immunoprecipitated with Argonaute proteins in vivo and were active in luciferase assays, indicating that they are functional. Bioinformatics analysis predicts substantial differences in targeting between miRNAs with minor 5’ differences and in support of this we report that a 5’ isomiR-9-1 gained the ability to inhibit the expression of DNMT3B and NCAM2 but lost the ability to inhibit CDH1 in vitro. This result was confirmed by the use of isomiR-specific sponges. Our analysis of the miRGator database indicates that a small percentage of human miRNA genes express isomiRs as the dominant transcript in certain cell types and analysis of miRBase shows that 5’ isomiRs have replaced canonical miRNAs many times during evolution. This strongly indicates that isomiRs are of functional importance and have contributed to the evolution of miRNA genes Sequence library of miRNAs from a single sample of human foetal mesenchymal stem cells. Results tested and confirmed by northern blotting. Please note that only raw data files are available for the embryonic and neual samples and thus, directly submitted to SRA (SRX547311, SRX548700, respectively under SRP042115/PRJNA247767)
Project description:To characterize the human plasma microtranscriptome profile at first trimester of pregnancy in presence or not of pregnancy complications, we sequenced microRNAs in plasma samples collected from pregnant women between the 4th and the 16th weeks of pregnancy. We then performed differential expression analyses to assess the miRNA profile diffrences according to the presence of pregnancy complications or not (i.e. Gestational diabetes mellitus, Gestational hypertension or preeclampsia vs. normal pregnancies).
Project description:Gestation-matched endometrium was collected from women with ectopic pregnancy (EP) (n=11) and intrauterine pregnancies (IUP) (n=13). RNA was extracted from the tissue. In addition, tissues were prepared for histological analysis for degree of decidualization. We compared a) the samples from EP that were decidualized (n=6) with non-decidualized samples (n=5), and b) the decidualized EP (n=6) with decidualization-matched IUP (n=6) samples.