Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Human exosomal placenta-specific miRNAs modulate expression of target genes in maternal immune cells during pregnancy

ABSTRACT: We investigated placenta-specific miRNA miR-517a (miR-517a-3p) functions in Jurkat cells. For this purpose, we identified candidate target mRNAs of the placenta-specific miRNAs using DNA microarray analysis. Transfection of Jurkat cells with miR-517a significantly downregulated 123 genes. Among the 123 genes identified, we searched for potential direct targets of miR-517a using the online software MicroCosm Targets. Seven genes, ALDH1B1, ANP32E, DHFR, FAT2, IGSF5, PRKG1, and RSPO3, had at least one potential miR-517a binding site in their 3M-bM-^@M-^Y-UTRs. Furthermore, we revealed that PRKG1, one of the seven genes, was a miR-517a target using an in vitro experimental validation system. We compared gene expression pattern of miR-517aM-bM-^@M-^Soverexpressing Jurkat cells with that of negative control.

ORGANISM(S): Homo sapiens

SUBMITTER: Kazuya Yuge

PROVIDER: E-GEOD-50814 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:The miR-29 family is an important player in the molecular pathophysiology of distinct types of cancer, with roles that seems to depend on cellular context. Reduced miR-29 levels are associated with more aggressive disease and its overexpression in cancer and leukemic cell lines inhibits proliferation. In contrast, its overexpression in hematopoietic progenitors, promotes leukemogenesis. We explored the potential roles of miR-29a in the molecular pathophysiology of T-cell acute lymphoblastic leukemia (T-ALL). As compared to normal T-cells, miR-29a levels are extremely reduced in T-ALL and in the Jurkat cell line. Microarrays analysis in Jurkat cells, following the introduction of synthetic miR-29a mimics, revealed the down-regulation of several predicted targets, including previously described targets (DNMT3a/b, CDK6, PXDN, MCL1, PIK3R1 and CXXC6), and novel targets with roles in active DNA demethylation, such as members of the ten-eleven-translocation (TET) family and TDG. Reduced miR-29a levels contribute to altered epigenetics, as its introduction in Jurkat cells, promotes the demethylation of the AHR gene (commonly methylated in T-ALL). In T-ALL patients, miR-29a levels are significantly associated with blast counts and disease free survival. Our results highlight the relevance of miR-29 in T-ALL physiopathology, and may help to clarify some contrasting findings reported in the literature. In order to identify potential miR-29a targets in T-cell lymphoblastic leukemia, Jurkat cells were electroporated with synthetic RNA molecules corresponding to miR-29a mimics (pre-miR) or inhibitors (anti-miR); and microarray profiles were compared to the profiles of Jurkat cells eletroporated with the corresponding negative controls. Transcripts with levels reduced following the introduction of the pre-miR-29a (as compared to pre-miR-Control), or transcripts acumulated folowing introduction of the anti-miR-29a inhibitor (as compared to anti-miR-Control), were considered potential targets; and were further compared to microRNA target prediction databases. Two paired samples were used for this analysis.

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Human exosomal placenta-specific miRNAs modulate expression of target genes in maternal immune cells during pregnancy

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