High-Throughput Sequencing Reveals Circulating miRNAs as Potential Biomarkers of Kidney Damage in Patients with Systemic Lupus Erythematosus.

Pierine España-Puccini,Lisneth Almendrales,Xu-Jie Zhou,Gustavo Aroca,Edwin Rico,Yirys Díaz-Olmos,Eduardo Egea,Hernan Lorenzi,Antonio Iglesias,Elkin Navarro-Quiroz,Roberto Navarro,Lisandro Pacheco-Lugo

doi:10.1371/journal.pone.0166202

Abstract

Renal involvement is one of the most severe manifestations of systemic lupus erythematosus (SLE). Renal biopsy is the gold standard when it comes to knowing whether a patient has lupus nephritis, and the degree of renal disease present. However, the biopsy has various complications, bleeding being the most common. Therefore, the development of alternative, non-invasive diagnostic tests for kidney disease in patients with SLE is a priority. Micro RNAs (miRNAs) are differentially expressed in various tissues, and changes in their expression have been associated with several pathological processes. The aim of this study was to identify changes in the abundance of miRNAs in plasma samples from patients with lupus nephritis that could potentially allow the diagnosis of renal damage in SLE patients. This is an observational case-control cross-sectional study, in which we characterized the differential abundance profiles of miRNAs among patients with different degrees of lupus compared with SLE patients without renal involvement and healthy control individuals. We found 89 miRNAs with changes in their abundance between lupus nephritis patients and healthy controls, and 17 miRNAs that showed significant variations between SLE patients with or without renal involvement. Validation for qPCR of a group of miRNAs on additional samples from lupus patients with or without nephritis, and from healthy individuals, showed that five miRNAs presented an average detection sensitivity of 97%, a specificity of 70.3%, a positive predictive value of 82.5%, a negative predictive value of 96% and a diagnosis efficiency of 87.9%. These results strongly suggest that miR-221-5p, miR-380-3p, miR-556-5p, miR-758-3p and miR-3074-3p are potential diagnostic biomarkers of lupus nephritis in patients with SLE. The observed differential pattern of miRNA abundance may have functional implications in the pathophysiology of SLE renal damage.

Highlights

Lupus nephritis (LN) is the commonest and one of the most serious manifestations of systemic lupus erythematosus (SLE) [1,2], with reports of 5-year renal survivals ranging from 46% to 95% with treatment [3]
LN activity was evaluated based on the Systemic Lupus Erithematous Disease Activity Index (SLEDAI) [13]
In order to keep remission, patients utilized oral steroids or oral steroids mixed with mycophenolate mofetil or azathioprine [12]. 10 plasma samples from lupus patients with no nephritis (LNN), 7 plasma samples from control healthy individuals (CTL), 4 plasma samples from LN class II (LNII), 4 plasma samples from LN class III (LNIII), and 6 plasma samples from LN class IV (LNIV) were used to high-throughput sequencing

Summary

Introduction

Lupus nephritis (LN) is the commonest and one of the most serious manifestations of SLE [1,2], with reports of 5-year renal survivals ranging from 46% to 95% with treatment [3]. Renal biopsy is the gold standard for providing information on the histological classes of LN and the relative degree of activity and chronicity in the glomeruli It is invasive and serial biopsies are impractical in the monitoring of LN. We evaluated the miRNAs circulating in plasma samples of individuals with different stages of renal involvement using a high throughput sequencing approach, and we found a group of miRNAs whose expression pattern correlates with renal involvement in patients with SLE. To our knowledge, this is the first study that has used a group of patients with different involvement of renal injury in SLE to compare circulating miRNAs using large-scale sequencing

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