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Abstract Number: 754

The Global miRNA Whole Blood Profile in Systemic Sclerosis and Its Correlation with Serum Cytokine Levels

Gloria Salazar1, Maureen Mayes2, John Hagan3, Minghua Wu1, John D. Reveille4,5 and Shervin Assassi1, 1Rheumatology, University of Texas Health Science Center at Houston, Houston, TX, 2Internal Medicine/Rheumatology, University of TX Health Science Center -Houston, Houston, TX, 3Neurosurgery, University of Texas at Houston, Houston, TX, 4Internal Medicine/Rheumatology, University of Texas Health Science Center at Houston, Houston, TX, 5Internal Medicine/Rheumatology, Univ of Texas Health Science Center at Houston, Houston, TX

Meeting: 2014 ACR/ARHP Annual Meeting

Keywords: Biomarkers, cytokines, Epigenetics and systemic sclerosis, MicroRNA

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Session Information

Session Title: Systemic Sclerosis, Fibrosing Syndromes and Raynaud's - Pathogenesis, Animal Models and Genetics

Session Type: Abstract Submissions (ACR)

The Global miRNA Whole Blood Profile in Systemic Sclerosis and its Correlation with Serum Cytokine Levels

Background/Purpose:

Several studies have implicated miRNAs in the pathogenesis of systemic sclerosis (SSc).1-3 Recent advances in quantitative polymerase chain reaction (qPCR) allow simultaneous measurement of hundreds of miRNAs. The objective of this study was to use this technology to identify the unbiased, global miRNA profiling of SSc whole blood and evaluate its correlation with plasma cytokine levels.

Methods:

We investigated the miRNA profile in SSc whole blood compared to unaffected controls using multiplex qPCR platform. We obtained blood samples from 10 patients with early SSc (≤5 yrs, on no immunosuppression) and 10 age-, gender- and ethnicity matched controls. Eight patients had diffuse disease and two had limited disease. The mean disease duration was 2.2 years. Levels of 752 miRNAs were determined. Unsupervised hierarchical clustering analysis was performed. Patient and control samples miRNA levels were compared and differences with a p<0.01, false discovery rate (FDR) <10% and fold change >2 were considered statistically significant.

A quantitative, multiplexed immunoassay designed to measure 45 cytokines, chemokines and acute-phase reactants was used to determine the serum cytokine levels (myriad human inflammationMAP) in order to correlate expression of miRNA with their predicted targets.

Results:

The comparison of patient to control whole blood samples revealed 16 miRNA that were differentially expressed (Table 1). All miRNAs, except for miR-10b-5p, were downregulated in SSc compared to controls. Notably four differentially expressed miRNA in whole blood originate from the same cluster located in 14q32.3. The unsupervised hierarchical clustering analysis of whole blood miRNA profile separated the two groups but three patients clustered along with controls.

In the cytokine analysis, MCP1 (CCL2), IL-10, MMP-9, TNFR2, VCAM1 and ICAM1 were differentially expressed in SSc patients. MiR-370 levels highly correlated with MCP-1 protein levels (rs= -0.6, p=0.004) which is a predicted target of this miRNA. 

Conclusion:

To our knowledge, this is the first global, unbiased examination of miRNA in SSc whole blood and the first correlation of miRNA with SSc plasma cytokine levels. The miRNA profile showed 16 miRNAs that are dysregulated in SSc whole blood. MiR-370 levels were differentially expressed in SSc blood and highly correlated with MCP-1 protein levels. We have previously reported also an upregulation of this miRNA in SSc skin.  Furthermore, miR-370  targeted TGFbR-II in two independent studies underscoring its potential role in SSc pathogenesis.

Table 1. SSc whole blood dysregulated miRNA

 

miRNA

Location

Fold Change

p

miR-1238-3p

19p13.2

0.18

<0.001

miR-506-3p

Xq27.3

0.33

<0.001

miR-134

14q32

0.42

0.001

miR-579

5p13.3

0.2

0.001

miR-144-3p

17q11.2

0.27

0.002

miR-320a

8p21.3

0.5

0.002

miR-370

14q32

0.16

0.003

miR-1537

1q42.3

0.3

0.003

miR-2113

6q16.1

0.19

0.004

miR-877-5p

6p21.33

0.48

0.004

miR-101-5p

1p31.3

0.33

0.004

miR-200b-5p

1p36.33

0.33

0.006

miR-511

10p12.33

0.26

0.007

miR-300

14q32

0.26

0.008

miR-584-5p

5q32

0.49

0.009

miR-10b-5p

2q31.1

5.39

0.005

 


Disclosure:

G. Salazar,
None;

M. Mayes,
None;

J. Hagan,
None;

M. Wu,
None;

J. D. Reveille,
None;

S. Assassi,
None.

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