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research
Expression of Regulatory Platelet MicroRNAs in Patients with Sickle Cell Disease
Authors
A Abdollahi
A Dixon-McIver
+70 more
A Osman
A Schedel
A Smolenski
A Tomer
AA Kondkar
AD Blann
AI Badeaux
AJ Saldanha
AM Healy
BM Steele
Claudia Coronnello
D Betel
D Betel
DA Hosack
DP Bartel
DV Gnatenko
EM Novelli
ER Popescu
G Bazzoni
Gregory J. Kato
Guoying Yu
H Bruchova
H Seitz
HA Brittain
I Hers
I Toma
J Villagra
Jen-Tsan Ashley Chi
JF Noronha
JS Mohan
K Joop
K Kaushansky
K Konishi
Kimberly Woodhouse
LL Horstman
LR Queen
M Kannan
M Kertesz
Maria G. Kapetanaki
Mark T. Gladwin
MC Ammons
ML Freedman
ML Jison
MP Hunter
N Chavda
N Raghavachari
Naftali Kaminski
Nalini Raghavachari
P Landry
Panayiotis V. Benos
PV Browne
R Antonucci
R Garzon
R Ross
R Visone
RC Friedman
RJ Berckmans
RL Nachman
RT Schermuly
S Amisten
S Kim
S Masaki
S Mendjan
S Nagalla
Shilpa Jain
SP Lee
Suchitra Barge
T Wun
V Ambros
W Huang da
Publication date
12 April 2013
Publisher
'Public Library of Science (PLoS)'
Doi
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on
PubMed
Abstract
Background: Increased platelet activation in sickle cell disease (SCD) contributes to a state of hypercoagulability and confers a risk of thromboembolic complications. The role for post-transcriptional regulation of the platelet transcriptome by microRNAs (miRNAs) in SCD has not been previously explored. This is the first study to determine whether platelets from SCD exhibit an altered miRNA expression profile. Methods and Findings: We analyzed the expression of miRNAs isolated from platelets from a primary cohort (SCD = 19, controls = 10) and a validation cohort (SCD = 7, controls = 7) by hybridizing to the Agilent miRNA microarrays. A dramatic difference in miRNA expression profiles between patients and controls was noted in both cohorts separately. A total of 40 differentially expressed platelet miRNAs were identified as common in both cohorts (p-value 0.05, fold change>2) with 24 miRNAs downregulated. Interestingly, 14 of the 24 downregulated miRNAs were members of three families - miR-329, miR-376 and miR-154 - which localized to the epigenetically regulated, maternally imprinted chromosome 14q32 region. We validated the downregulated miRNAs, miR-376a and miR-409-3p, and an upregulated miR-1225-3p using qRT-PCR. Over-expression of the miR-1225-3p in the Meg01 cells was followed by mRNA expression profiling to identify mRNA targets. This resulted in significant transcriptional repression of 1605 transcripts. A combinatorial approach using Meg01 mRNA expression profiles following miR-1225-3p overexpression, a computational prediction analysis of miRNA target sequences and a previously published set of differentially expressed platelet transcripts from SCD patients, identified three novel platelet mRNA targets: PBXIP1, PLAGL2 and PHF20L1. Conclusions: We have identified significant differences in functionally active platelet miRNAs in patients with SCD as compared to controls. These data provide an important inventory of differentially expressed miRNAs in SCD patients and an experimental framework for future studies of miRNAs as regulators of biological pathways in platelets. © 2013 Jain et al
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