MicroRNA modulation in complex regional pain syndrome

Irina A Orlova, Guillermo M Alexander, Rehman A Qureshi, Ahmet Sacan, Alessandro Graziano, James E Barrett, Robert J Schwartzman, Seena K Ajit, Irina A Orlova, Guillermo M Alexander, Rehman A Qureshi, Ahmet Sacan, Alessandro Graziano, James E Barrett, Robert J Schwartzman, Seena K Ajit

Abstract

Background: Aberrant expression of small noncoding RNAs called microRNAs (miRNAs) is a common feature of several human diseases. The objective of the study was to identify miRNA modulation in patients with complex regional pain syndrome (CRPS) a chronic pain condition resulting from dysfunction in the central and/or peripheral nervous systems. Due to a multitude of inciting pathologies, symptoms and treatment conditions, the CRPS patient population is very heterogeneous. Our goal was to identify differentially expressed miRNAs in blood and explore their utility in patient stratification.

Methods: We profiled miRNAs in whole blood from 41 patients with CRPS and 20 controls using TaqMan low density array cards. Since neurogenic inflammation is known to play a significant role in CRPS we measured inflammatory markers including chemokines, cytokines, and their soluble receptors in blood from the same individuals. Correlation analyses were performed for miRNAs, inflammatory markers and other parameters including disease symptoms, medication, and comorbid conditions.

Results: Three different groups emerged from miRNA profiling. One group was comprised of 60% of CRPS patients and contained no control subjects. miRNA profiles from the remaining patients were interspersed among control samples in the other two groups. We identified differential expression of 18 miRNAs in CRPS patients. Analysis of inflammatory markers showed that vascular endothelial growth factor (VEGF), interleukin1 receptor antagonist (IL1Ra) and monocyte chemotactic protein-1 (MCP1) were significantly elevated in CRPS patients. VEGF and IL1Ra showed significant correlation with the patients reported pain levels. Analysis of the patients who were clustered according to their miRNA profile revealed correlations that were not significant in the total patient population. Correlation analysis of miRNAs detected in blood with additional parameters identified miRNAs associated with comorbidities such as headache, thyroid disorder and use of narcotics and antiepileptic drugs.

Conclusions: miRNA profiles can be useful in patient stratification and have utility as potential biomarkers for pain. Differentially expressed miRNAs can provide molecular insights into gene regulation and could lead to new therapeutic intervention strategies for CRPS.

Figures

Figure 1
Figure 1
Differential expression of miRNAs in blood from CRPS patients and control samples. A clustergram of the samples and miRNAs identifying sample subpopulations with respect to the significant differentially expressed miRNAs. A total of 60% of the patients (Group 3) showed differential regulation for these miRNAs (mostly downregulation), whereas the rest of the patients were more heterogeneous and clustered together with the control samples. P and C represent patients and controls, respectively. Red, high; black, average; green, low.
Figure 2
Figure 2
Elevation of inflammatory markers in CRPS patients. VEGF, IL1Ra, and MCP1 showed significant differential expression in CRPS patients vs. control samples, with p values 0.0002, 0.0004, and 0.0005, respectively. The error bars show the standard error of the means.
Figure 3
Figure 3
Circos diagram showing the correlation of selected parameters and miRNAs. The Circos diagram showing the correlation of selected medical conditions with other clinical parameters and miRNAs. The nodes along the circle are colored by the total strength of correlation of the corresponding variable, such that variables with many strong correlations are shown in red. The links between variables indicate their correlation. Only correlations with adjusted p value less than 0.01 are shown. Groups of miRNAs showing similar correlations with other variables are labeled as "miRNAs" for brevity. The links are colored by the Spearman's correlation value, with strong negative correlations shown in darker blue (e.g., narcotics vs. hsa-miR-191) and strong positive correlations shown in darker red (e.g., pain level vs. "IL1ra, VEGF, miRNAs"), as implicated by the color-bar. See the Supplementary table 1 for a complete list of correlations.

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