Project description:Preeclampsia (PE), which affects 2-7% of human pregnancies, causes significant maternal and neonatal morbidity and mortality. To better understand the pathophysiology of PE, gene expression profiling of placental tissue from 5 controls and 5 PEs were assessed using microarray. A total of 224 transcripts were identified as being significantly differentially expressed (fold change > 2 and q value < 0.05 in the SAM software), GO enrichment analysis indicated that genes involved hypoxia, oxidative and reductive processes were significantly changed. Ten differentially expressed genes (DEGs) involved in these biological process were further verified by quantitative real-time PCR. Finally, the potential therapeutic agents for PE were explored via Connectivity Map database . In conclusion, the data obtained in this study might provide clues to better understand the pathophysiology of PE and found potential therapeutic agents for PE patients. gene expression profiling of placental tissue from 5 controls and 5 PEs were assessed using microarray.
Project description:Preeclampsia (PE), which affects 2-7% of human pregnancies, causes significant maternal and neonatal morbidity and mortality. To better understand the pathophysiology of PE, gene expression profiling of placental tissue from 5 controls and 5 PEs were assessed using microarray. A total of 224 transcripts were identified as being significantly differentially expressed (fold change > 2 and q value < 0.05 in the SAM software), GO enrichment analysis indicated that genes involved hypoxia, oxidative and reductive processes were significantly changed. Ten differentially expressed genes (DEGs) involved in these biological process were further verified by quantitative real-time PCR. Finally, the potential therapeutic agents for PE were explored via Connectivity Map database . In conclusion, the data obtained in this study might provide clues to better understand the pathophysiology of PE and found potential therapeutic agents for PE patients.
Project description:Pre-eclampsia is one of the most serious pregnancy-associated disorders, and is defined by hypertension (systolic blood pressure higher than 140mmHg or diastolic blood pressure higher than 90mmHg) with proteinuria (more than 0.3g/day). It is not a simple complication of pregnancy, but is rather a syndrome of multiple organ failure involving the liver, kidney, and lung, as well as coagulatory and neural systems. There is now an emerging consensus that pre-eclampsia is a complex polygenetic trait in which maternal and fetal genes, as well as environmental factors, are involved, although the precise mechanism of the disorder has remained elusive. In this study, we have performed gene expression profiling to further elucidate the mechanisms underlying the development of the pre-eclamptic condition. We thus compared the expression profiles in placentas from women who underwent a normal pregnancy and from women suffering from severe pre-eclampsia. In addition, we compared the expression data between the early and late onset forms of pre-eclampsia and obtained a number of potential new prognostic biomarkers for this disease. Experiment Overall Design: Placental biopsies were obtained during Caesarean section from both normotensive patients and from those with pre-eclampsia (n = 14) (early onset type; earlier than 31 weeks gestation, n = 6 and late onset type; 31 weeks gestation or later, n = 8). Pre-eclampsia was defined as a blood pressure of higher than 160/110 mmHg, with proteinuria of more than 2g in a 24 hour collection. The expression profiles of approximately 47,669 genes were analyzed using a Whole Human Genome Oligo Microarray Kit (Agilent Technologies). A total of 10 placentas from women with pre-eclampsia and four from normal subjects were used in the hybridizations. Reverse transcription labeling and hybridization was performed using the protocol recommended by the manufacturer. The microarray experiments were then carried out using competitive hybridization experiments with Cy3 and Cy5-labeled targets; one with test placental RNAs from patients or normal control subjects, and another using pooled normal placental RNA as a template control for normalization. The glass slides were scanned using an Agilent G2565BA microarray scanner. Scanned images were then analyzed using Feature Extraction software. The average signal intensities were corrected for median background intensity and transferred with GenBank descriptors to a Microsoft Excel data spreadsheet (Microsoft, Redmond, WA). Data analysis was performed using Genespring software version 6.1 (Silicon Genetics, Redwood City, CA). After intensity dependent normalization (Lowess), the expression levels relative to the control were calculated as a ratio, and the expression profiles were then compared between each pre-eclamptic or normal sample.
Project description:Maternal Blood histamine levels are tightly controlled in normal pregnancy. However, in specific complications of human pregnancy such as pre-eclampsia the levels of both placental and maternal blood histamine increase. Increasing blood histamine levels nonetheless, have been associated with oxidative stress, endothelial dysfunction, abnormal tissue growth, and Th1/TH2 imbalance, which are also linked to pre-eclampsia. Little is known of the molecular responses in the placenta to the prolonged exposure to increasing histamine levels in the presence of changing oxygen concentrations. We used microarray to detail the global programme of placental gene expression in response to histamine and oxygen and identified distinct classes of regulated genes underlying the molecular functions of histamine in the placenta.
Project description:Genome-wide analysis of decidual transcriptome in pre-eclampsia compared with normotensive controls to find differentially expressed genes/pathways. Preeclampsia (PE) is a common and serious pregnancy hypertensive disorder with a strong genetic component. The study aims were to use genome-wide analysis of decidual transcriptome in pre-eclampsia compared with normotensive controls to identify differentially expressed biological pathways, and to determine common pathways between the transcriptome and previously identified maternal susceptibility genes.
Project description:Genome-wide analysis of decidual transcriptome in pre-eclampsia compared with normotensive controls to find differentially expressed genes/pathways. Preeclampsia (PE) is a common and serious pregnancy hypertensive disorder with a strong genetic component. The study aims were to use genome-wide analysis of decidual transcriptome in pre-eclampsia compared with normotensive controls to identify differentially expressed biological pathways, and to determine common pathways between the transcriptome and previously identified maternal susceptibility genes. Total RNA from decidua basalis obtained from pre-eclamptic and normotensive control patients at Caesarean section
Project description:To determine differential expression of microRNAs in placentae with severe pre-eclampsia and normal placenta. Differential expression of microRNAs in placentae (4 severe pre-eclampsia and 4 normal control) was screened by microarray platform, then some differential microRNAs were selected and validated with real-time quantitative reverse transcription polymerase chain reaction in placentae of severe pre-eclampsia (n=24) and normal control group (n=26). Results: We validated the expression of some microRNAs altered in the microarray, and found the following microRNAs were significantly increased in severe pre-eclampsia placentae: miR-16, miR-29b, miR-195, miR-26b, miR-181a, miR-335 and miR-222. Conclusion: These differential microRNAs may play an important role in pathogenesis of pre-eclampsia.
Project description:We report abnormal pregnancy outcomes in mice lacking NKG2A, including altered placental transcription. In humans, we find HLAB-21T snp to be associated with an increased risk of pre-eclampsia.
Project description:Background: We have previously used the rat 4 day Complete Freund's Adjuvant (CFA) model to screen compounds with potential to reduce osteoarthritic pain. The aim of this study was to identify genes altered in this model of osteoarthritic pain and use this information to infer analgesic potential of compounds based on their own gene expression profiles using the Connectivity Map approach. Results: Using microarrays, we identified differentially expressed genes in L4 and L5 dorsal root ganglia (DRG) from rats that had received intraplantar CFA for 4 days compared to matched, untreated control animals. Analysis of these data indicated that the two groups were distinguishable by differences in genes important in immune responses, nerve growth and regeneration. This list of differentially expressed genes defined a âCFA signatureâ. We used the Connectivity Map approach to identify pharmacologic agents in the Broad Institute Build02 database that had gene expression signatures that were inversely related (ânegatively connectedâ) with our CFA signature. To test the predictive nature of the Connectivity Map methodology, we tested phenoxybenzamine (an alpha adrenergic receptor antagonist) â one of the most negatively connected compounds identified in this database - for analgesic activity in the CFA model. Our results indicate that at 10mg/kg, phenoxybenzamine demonstrated analgesia comparable to that of Naproxen in this model. Conclusion: Evaluation of phenoxybenzamine-induced analgesia in the current study lends support to the utility of the Connectivity Map approach for identifying compounds with analgesic properties in the CFA model. A naive (control) group of rats (n = 6) and a group of rats injected with CFA (n = 6) were used for gene expression profiling experiments. One animal from the control group did not yield sufficient amount of RNA for microarray and thus was omitted from further processing. In total, 5 microarrays each from the 5 animals in the control group, and 6 microarrays each from the 6 animals in the CFA group were analyzed.
Project description:To explore the mechanism underlying anti-leukaemia effect of sodium valproate, the growth and survival of the K562 cell line were investigated using the Annexin V/PI dual staining method. Global profiles of gene expression in K562 cells exposed to sodium valproate were assessed using gene expression microarray and validated by qRT-PCR. Gene ontology analysis was performed to establish the function of differentially expressed genes. The differentially expressed genes identified by using gene expression array were further used to query the CMAP database to retrieve a ranked list of compounds that act on the same intracellular targets as sodium valproate. A significant increase in cell apoptosis was observed in valproate exposed K562 cells using flow cytometry. A total of 706 transcripts were identified as being significantly differentially expressed. Eight differentially expressed genes that might be involved in apoptosis were verified by qRT-PCR. The significant enrichment analysis of gene ontology terms for the differentially expressed genes revealed that these genes are involved in many important biological processes such as apoptosis. The connectivity map analysis showed gene expression profile in K562 cells exposed to sodium valproate observed in this study was most similar to that of HDACi and PI3K inhibitor in the connectivity map database, suggesting that sodium valproate might exert anti-leukaemic action by inhibiting HDAC as well as inhibiting PI3K pathway. In conclusion, the data obtained in this study might provide the ground for further studies to elucidate the molecular and therapeutic potential of VPA in leukaemia treatment. Gene expression in K562 cell line were measured after exposure to 2 mM valproate for 12 hours or left untreated as a control using Agilent SurePrint G3 Human GE 8x60K Microarray.