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| 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
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| 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.
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| 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.
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| 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
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| Phone |
+390592058065
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| Organization name |
Centre for Regenerative Medicine
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| Department |
Life Sciences
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| Street address |
Via Gottardi 100
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| City |
Modena |
| ZIP/Postal code |
41100 |
| Country |
Italy |
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| Platforms (1) |
| GPL13667 |
[HG-U219] Affymetrix Human Genome U219 Array |
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| Samples (6)
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| Relations |
| BioProject |
PRJNA350499 |
| 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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