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MRNA profiling of human Multipotent Adipose-Derived Stem (hMADS) cells transfected with miR-29a and subjected to adipocyte differentiation

ABSTRACT: The human microRNA hsa-miR-29a is a member of the miR-29 family of microRNAs, which is expressed in the adipogenic lineage. To identify putative direct target mRNAs of the miR-29 family, and to get an idea about potential functions of the miR-29 family in adipocyte development, we transfected human Multipotent Adipose-Derived Stem (hMADS) cells with miR-29a mimics (leading to elevation of intracellular levels of mature miR-29a). Subsequently, gene expression profiling was performed for hMADS cells at day 0 (48 h after transfection) and at day 3, i.e. 3 days after adipocyte differentiation was induced by a chemically defined medium.

ORGANISM(S): Homo sapiens

PROVIDER: GSE65704 | GEO | 2019/01/23

REPOSITORIES: GEO

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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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MRNA profiling of human Multipotent Adipose-Derived Stem (hMADS) cells transfected with miR-29a and subjected to adipocyte differentiation

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