Transcriptomics,Genomics

Dataset Information

47

Gene expression profile (GEP) of miR-34a-5p-overexpressing CD34+ HPCs


ABSTRACT: Primary Myelofibrosis (PMF) is a myeloproliferative neoplasm characterized by hyperplastic megakaryopoiesis and myelofibrosis. Through a gene expression profile (GEP) study we recently highlighted the upregulationof miR-34a-5p in PMF versus healthy donor (HD) CD34+ hematopoietic progenitor cells (HPCs). To shed some light into the role of miR-34a-5p in PMF pathogenesis, here we unravelled the effects of the overexpression of miR-34a-5p in HPCs forcing its expression in HPCs. We showed that enforced expression of miR-34a-5p blocks proliferation and favours the megakaryocyte and monocyte/macrophage commitment of HPCs. Interestingly, we identified lymphoid enhancer-binding factor 1 (LEF1) and nuclear receptor subfamily 4, group A, member 2 (NR4A2) transcripts as miR-34a-5p-targets downregulated after miR-34a-5p overexpression in HPCs as well as in PMF compared with HD HPCs. Remarkably, the knockdown of NR4A2 in HPCs mimicked the antiproliferative effects of miR-34a-5p overexpression, while the silencing of LEF1 phenocopied the effects of miR-34a-5p overexpression in HPCs lineage choice, by stimulating the megakaryocyte and monocyte/macrophage commitment. Overall design: Human CD34+ cells were transfected by using the 4D-Nucleofector™ System (Lonza). Briefly, starting from the day after CD34+ cells purification, each sample was electroporated twice, once every 24 hours, with 3 µg of mirVana™ miR-34a-5p mimic or mirVana™ miRNA mimic Negative Control (NegCTR mimic). For each electroporation, 4×105 CD34+ cells were resuspended in 100 µL of P3 Primary Cell Solution (Lonza) containing 3 µg of miR-34a-5p mimic or NegCTR mimic and pulsed with the program DS112. Gene expression profile (GEP) was performed on total RNA derived from three independent experiments at 24h after the last nucleofection.

INSTRUMENT(S): [HG-U219] Affymetrix Human Genome U219 Array

SUBMITTER: Rossella Manfredini  

PROVIDER: GSE89156 | GEO | 2017-03-08

SECONDARY ACCESSION(S): PRJNA350499

REPOSITORIES: GEO

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Publications

Role of miR-34a-5p in Hematopoietic Progenitor Cells Proliferation and Fate Decision: Novel Insights into the Pathogenesis of Primary Myelofibrosis.

Bianchi Elisa E   Ruberti Samantha S   Rontauroli Sebastiano S   Guglielmelli Paola P   Salati Simona S   Rossi Chiara C   Zini Roberta R   Tagliafico Enrico E   Vannucchi Alessandro Maria AM   Manfredini Rossella R  

International journal of molecular sciences 20170113 1


Primary Myelofibrosis (PMF) is a chronic Philadelphia-negative myeloproliferative neoplasm characterized by a skewed megakaryopoiesis and an overproduction of proinflammatory and profibrotic mediators that lead to the development of bone marrow (BM) fibrosis. Since we recently uncovered the upregulation of miR-34a-5p in PMF CD34+ hematopoietic progenitor cells (HPCs), in order to elucidate its role in PMF pathogenesis here we unravelled the effects of miR-34a-5p overexpression in HPCs. We showed  ...[more]

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