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| Status |
Public on Dec 31, 2013 |
| Title |
Global miRNA expression status of human hematopoietic stem–progenitor cells modulates myelopoiesis |
| Organism |
Homo sapiens |
| Experiment type |
Non-coding RNA profiling by high throughput sequencing
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| Summary |
We investigated how suppression of the most upstream microRNA–processing RNase, Drosha, affects the differentiation of human CD34+ hematopoietic stem–progenitor cells (HSPCs). We hypothesized that knock-down of Drosha would alter blood lineage development by modulating the expression of microRNAs. Lentiviral delivery to HSPCs of a short-hairpin targeting Drosha resulted in a viable phenotype with promotion of myeloid, and especially monocytic, maturation and suppression of apoptosis. Our results show that Drosha deficiency triggered a parallel upregulation of components of the RNAi machinery, including DGCR8, Dicer and Ago2. Deep sequencing analyses revealed global miRNA deficiency after Drosha short-hairpin treatment with relative maintenance of mature miR-223 expression. Restoration of miR-223 to normal levels after Drosha knock-down further enhanced monocytic maturation concomitant with the modulation of myeloid transcription factors that promoted monocytic differentiation. Our results support a miRNA accentuation model in which relative enhancement of miR-223 increases levels of PU.1 thereby promoting monocytic differentiation.
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| Overall design |
CD34+ HSPCs were isolated from umbilical cord blood and transduced with an empty lentivector (EV) or a lentivector encoding a short-hairpin RNA targeting the pri-miRNA–processing enzyme, Drosha (shDrosha). EV and shDrosha transduced HSPCs were grown in liquid culture promoting myelopoiesis and sampled on days 0 and 7 for total RNA collection. Total RNA was size fractionated to enrich for the small RNA population and deep sequenced using ABI's SOLiD 4.0 platform.
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| Contributor(s) |
Azizian KT, Rossi JJ |
| Citation missing |
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| Submission date |
May 30, 2013 |
| Last update date |
May 15, 2019 |
| Contact name |
Xiwei Wu |
| E-mail(s) |
xwu@coh.org
|
| Organization name |
City of Hope National Medical Center
|
| Department |
Computational and Quantitative Medicine
|
| Street address |
1500 E. Duarte Rd.
|
| City |
Duarte |
| State/province |
CA |
| ZIP/Postal code |
91010 |
| Country |
USA |
| |
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| Platforms (1) |
| GPL13393 |
AB SOLiD 4 System (Homo sapiens) |
|
| Samples (8)
|
| GSM1151387 |
short-hairpin Drosha (shDr) day 0 (D0) rep2 |
| GSM1151388 |
empty vector (EV) day 7 myeloid (D7 GMM) rep1 |
| GSM1151389 |
empty vector (EV) day 7 myeloid (D7 GMM) rep2 |
| GSM1151390 |
short-hairpin Drosha (shDr) day 7 myeloid (D7 GMM) rep1 |
| GSM1151391 |
short-hairpin Drosha (shDr) day 7 myeloid (D7 GMM) rep2 |
|
| Relations |
| BioProject |
PRJNA205943 |
| SRA |
SRP023502 |
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