Urinary Sediment mRNA Level of CREBBP and CYBA in Children With Steroid-Resistant Nephrotic Syndrome.

Wei Li,Lin Li,Lin He,Wenqing Xiang,Haidong Fu,Jianhua Mao,Xinyi Shou

doi:10.3389/fimmu.2021.801313

Abstract

BackgroundThis study aimed to evaluate gene expression patterns in urinary sediment samples of children with steroid-resistant nephrotic syndrome (SRNS).MethodsThe messenger RNA (mRNA) levels of 770 immune-related genes were detected using a NanoString nCounter platform. To verify the NanoString results, quantitative analysis of nine gene mRNAs was performed using real-time RT-PCR in more samples.ResultsFirstly, compared with the steroid-sensitive nephrotic syndrome (SSNS) group (n=3), significant changes were observed in the mRNA level of 70 genes, including MAP3K14, CYBA, SLC3A2, CREB-binding protein (CREBBP), CD68, forkhead box P1 (FOXP1), CD74, ITGB2, IFI30, and so forth, in the SRNS group (n=3). A total of 129 children with idiopathic nephrotic syndrome (INS), 15 with acute glomerulonephritis, and 6 with immunoglobulin A nephropathy (IgAN) were enrolled to verify the NanoString results. Compared with patients with IgAN, those with INS had significantly lower levels of FOXP1 (P=0.047) and higher levels of CREBBP (P=0.023). Among SSNS, the mRNA level of ITGB2 was significantly lower in the non-relapse group than in the non-frequent relapse and frequent-relapse groups (P=0.006). Compared with the SSNS group, CREBBP was significantly elevated in the SRNS group (P=0.02). Further, CYBA significantly decreased in the SRNS group (P=0.01). The area under the curve (AUC) for CREBBP and CYBA was 0.655 and 0.669, respectively. CREBBP had a sensitivity of 83.3% and a specificity of 49.4% and CYBA had a sensitivity of 58.3% and a specificity of 83.1% to rule out SSNS and SRNS. The diagnosis value was better for CREBBP+CYBA than for CREBBP or CYBA alone, indicating that the combination of CREBBP and CYBA was a more effective biomarker in predicting steroid resistance (AUC=0.666; sensitivity=63.9%; specificity=76.4%).ConclusionsThis study was novel in investigating the urinary sediment mRNA level in children with INS using high-throughput NanoString nCounter technology, and 70 genes that may relate to SRNS were found. The results revealed that the urinary sediment mRNA level of ITGB2 was significantly lower in the non-relapse group than in the non-frequent relapse and frequent-relapse groups. Meanwhile, CREBBP was significantly elevated and CYBA was significantly lowered in the SRNS group compared with the SSNS group.

Highlights

Idiopathic nephrotic syndrome (INS) is the most common glomerular disease among children
A set of baseline biopsies were performed in six patients with INS without steroid therapy enrolled in this study, including three with sensitive nephrotic syndrome (SSNS) and three with steroid-resistant nephrotic syndrome (SRNS), to define immune-related gene expression signatures associated with response to steroids in children with INS
A one-sided t test was conducted to rank top genes associated with SSNS and SRNS, using a custom panel of 770 genes on a NanoString nCounter platform

Summary

Introduction

Idiopathic nephrotic syndrome (INS) is the most common glomerular disease among children. It is characterized by massive proteinuria, hypoalbuminemia, and edema [1]. Childhood INS is most commonly caused by one of two idiopathic diseases: minimal change disease (MCD) and focal segmental glomerulosclerosis (FSGS) [2]. The pathogenesis of INS is unclear but related to autoimmune dysfunction, previous studies suggested that T cell dysfunction is one of the important mechanisms of MCD incidence [5, 6]. Many studies described that various alterations in cytokine production in process of INS, such as IL4, IL-13, TNF-a [7,8,9]. This study aimed to evaluate gene expression patterns in urinary sediment samples of children with steroid-resistant nephrotic syndrome (SRNS)

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