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Series GSE89156 Query DataSets for GSE89156
Status Public on Mar 08, 2017
Title Gene expression profile (GEP) of miR-34a-5p-overexpressing CD34+ HPCs
Organism Homo sapiens
Experiment type Expression profiling by array
Summary 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.
 
Contributor(s) Bianchi E, Ruberti S, Rontauroli S, Norfo R, Pennucci V, Rossi C, Zini R, Salati S, Ferrari S, Tagliafico E, Manfredini R
Citation(s) 28098757
Submission date Oct 25, 2016
Last update date Mar 21, 2019
Contact name Rossella Manfredini
E-mail(s) manfredini.rossella@unimore.it
Phone +390592058065
Organization name Centre for Regenerative Medicine
Department Life Sciences
Street address Via Gottardi 100
City Modena
ZIP/Postal code 41100
Country Italy
 
Platforms (1)
GPL13667 [HG-U219] Affymetrix Human Genome U219 Array
Samples (6)
GSM2359824 CD34+ miR34a mimic donor 1
GSM2359825 CD34+ miR34a mimic donor 2
GSM2359826 CD34+ miR34a mimic donor 3
Relations
BioProject PRJNA350499

Download family Format
SOFT formatted family file(s) SOFTHelp
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Series Matrix File(s) TXTHelp

Supplementary file Size Download File type/resource
GSE89156_RAW.tar 12.5 Mb (http)(custom) TAR (of CEL)
Processed data included within Sample table

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