Session Information
Date: Monday, November 6, 2017
Session Type: ACR Poster Session B
Session Time: 9:00AM-11:00AM
Background/Purpose: Small noncoding RNAs (sRNAs), such as microRNAs (miRNAs), are regulators of biological processes and serve as important biomarkers of disease. Most previous plasma sRNA studies in inflammatory autoimmune diseases have relied on PCR or microarrays to quantify candidate miRNAs based on limited information about their function; however, unbiased sRNA sequencing could reveal new miRNA markers of disease. We hypothesized that plasma miRNAs are altered in patients with rheumatoid arthritis (RA) compared to control subjects and some may serve as markers of RA diagnosis.
Methods: This cross-sectional study included patients with 167 patients with RA, and 91 age, race and sex matched control subjects. Sequencing was done by Illumina NextSeq500. High quality reads were mapped to the human genome using Bowtie1. MiRBase21.0 was used to quantify miRNAs. miRNA reads were compared between RA and control subjects by DESeq2 with adjustment for age, race sex, and batch with 5% false discovery rate and multiple test correction by Benjamini and Hochberg method, and separately by random forest analysis. The top 12 differentially expressed miRNAs were validated by qPCR. A miRNA qPCR panel for RA diagnosis was developed using logistic regression, whose discrimination capacity was assessed by area under the receiver operative characteristic curve (AUC).
Results: Of 262 reliably mapped microRNAs, 107 were significantly altered >1.5-fold in patients with RA. Among the top 12 differentially expressed miRNAs identified by both DESeq2 and random forest analysis, miR-22-3p, miR-22-5p, miR-24-3p, miR-29c-3p, miR-30e-5p, miR-130a-3p, miR-140-3p, miR-221-3p, and miR-345-5p remained significantly altered and reliably detectable when validated by qPCR (Table). A panel including miR-22-3p, miR-24-3p, and miR-140-3p had AUC=0.81 for distinguishing between RA and control subjects. This panel remained robust distinguishing between control subjects and both seronegative RA (AUC=0.85) and seropositive RA (AUC=0.79).
Conclusion: Several plasma miRNAs identified by sRNA sequencing are altered in patients with RA compared to control subjects and may serve as novel diagnostic markers of RA. Further validation is necessary to confirm these findings.
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Table. Top differentially expressed plasma miRNAs validated by qPCR. |
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|
RA (fM) |
Control (fM) |
Fold difference |
P |
|
miR-22-3p |
58.10 [45.30, 74.50] |
15.10 [10.80, 21.00] |
3.85 |
8.45E-12
|
|
miR-22-5p |
1.18 [0.89, 1.56] |
0.39 [0.25, 0.61] |
2.99 |
1.25E-07
|
|
miR-24-3p |
3.98 [2.95,5.37] |
1.32 [0.85, 2.06] |
3.01 |
1.19E-06
|
|
miR-29c-3p |
1.46 [1.18, 1.80] |
0.56 [0.41, 0.77] |
2.59 |
5.32E-09
|
|
miR-30e-5p |
4.23 [3.51, 5.11] |
3.24 [2.49, 4.21] |
1.31 |
2.39E-02
|
|
miR-130a-3p |
5.00 [3.51, 7.12] |
1.27 [0.81, 1.97] |
3.94 |
8.56E-08
|
|
miR-140-3p |
1.22 [1.00, 1.49] |
0.25 [0.17, 0.37] |
4.93 |
2.12E-13 |
|
miR-221-3p |
9.96 [7.48, 13.20] |
3.04 [2.15, 4.31] |
3.27 |
6.78E-09
|
|
miR-345-5p |
0.06 [0.04, 0.09] |
0.02 [0.01, 0.03] |
3.62 |
1.72E-04
|
|
*The geometric mean [interquartile range] is presented based on qPCR adjusted for triple spike-in control. |
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To cite this abstract in AMA style:
Ormseth MJ, Solus JF, Sheng Q, Guo Y, Ye F, Allen R, Vickers K, Stein CM. Small RNA Sequencing Identifies Plasma microRNA Panel for Rheumatoid Arthritis Diagnosis [abstract]. Arthritis Rheumatol. 2017; 69 (suppl 10). https://acrabstracts.org/abstract/small-rna-sequencing-identifies-plasma-microrna-panel-for-rheumatoid-arthritis-diagnosis/. Accessed .« Back to 2017 ACR/ARHP Annual Meeting
ACR Meeting Abstracts - https://acrabstracts.org/abstract/small-rna-sequencing-identifies-plasma-microrna-panel-for-rheumatoid-arthritis-diagnosis/