Construction and Analysis of Immune Infiltration-Related ceRNA Network for Kidney Stones

Abstract

Purpose: Kidney stones is a common medical issue that mediates kidney injury and even kidney function loss. However, the exact pathogenesis still remains unclear. This study aimed to explore the potential competing endogenous RNA (ceRNA)-related pathogenesis of kidney stones and identify the corresponding immune infiltration signature.Methods: One mRNA and one long non-coding RNA (lncRNA) microarray dataset was obtained from the GEO database. Subsequently, we compared differentially expressed mRNAs (DE-mRNAs) and lncRNAs between Randall’s plaques in patients with calcium oxalate (CaOx) stones and controls with normal papillary tissues. lncRNA-targeted miRNAs and miRNA–mRNA pairs were predicted using the online databases. lncRNA-related DE-mRNAs were identified using the Venn method, and GO and KEGG enrichment analyses were subsequently performed. The immune-related lncRNA–miRNA–mRNA ceRNA network was developed. The CIBERSORT algorithm was used to estimate the rate of immune cell infiltration in Randall’s plaques. The ceRNA network and immune infiltration were validated in the glyoxylate-induced hyperoxaluric mouse model and oxalate-treated HK-2 cells.Results: We identified 2,340 DE-mRNAs and 929 DE-lncRNAs between Randall’s plaques in patients with CaOx stones and controls with normal papillary tissues. lncRNA-related DE-mRNAs were significantly enriched in extracellular matrix organization and collagen-containing extracellular matrix, which were associated with kidney interstitial fibrosis. The immune-related ceRNA network included 10 lncRNAs, 23 miRNAs, and 20 mRNAs. Moreover, we found that M2 macrophages and resting mast cells were differentially expressed between Randall’s plaques and normal tissues. Throughout kidney stone development, kidney tubular injury, crystal deposition, collagen fiber deposition, TGF-β expression, infiltration of M1 macrophages, and activation of mast cells were more frequent in glyoxylate-induced hyperoxaluric mice compared with control mice. Nevertheless, M2 macrophage infiltration increased in early stages (day 6) and decreased as kidney stones progressed (day 12). Furthermore, treatment with 0.25 and 0.5 mM of oxalate for 48 h significantly upregulated NEAT1, PVT1, CCL7, and ROBO2 expression levels and downregulated hsa-miR-23b-3p, hsa-miR-429, and hsa-miR-139-5p expression levels in the HK-2 cell line in a dose-dependent manner.Conclusion: We found that significant expressions of ceRNAs (NEAT1, PVT1, hsa-miR-23b-3p, hsa-miR-429, hsa-miR-139-5p, CCL7, and ROBO2) and infiltrating immune cells (macrophages and mast cells) may be involved in kidney stone pathogenesis. These findings provide novel potential therapeutic targets for kidney stones.

CCL7 and olfactory transduction pathway activation play an important role in the formation of CaOx and CaP kidney stones

“…Among 92 patients with kidney stone who underwent percutaneous nephrolithotomy, CCL7 expression was markedly enhanced in the papillary tips versus urine samples (Sun et al, 2018). In another study, treatment with oxalate upregulated CCL7 expression in human renal proximal tubular epithelial cells (Xia et al, 2021). Therefore, CCL7 may contribute to the development of both CaOx and CaP stones.…”

The SOX4/EZH2/SLC7A11 signaling axis mediates ferroptosis in calcium oxalate crystal deposition-induced kidney injury

“…Finally, CAG-creER-EZH2 fl/fl (EZH2iKO) mice were constructed and injected with tamoxifen (75 mg/kg bodyweight) at week 6 to obtain EZH2 knockout mice. Mice were injected with glyoxylic acid (Gly; 120 mg/kg bodyweight; G10601; Sigma–Aldrich) for 12 days to create the kidney stone mice [ 28 , 29 ]. The impact of ferroptosis inhibition on CaOx-induced kidney injury was examined by injecting mice with liproxstatin-1 (Lip-1; 10 mg/kg bodyweight; S7699; Selleck) for 15 days [ 30 ].…”

CCL7 as a novel inflammatory mediator in cardiovascular disease, diabetes mellitus, and kidney disease

“…In response to acute kidney injury, B cells produce CCL7 to facilitate neutrophil and monocyte recruitment to the injured sites [69]. Treatment with oxalate upregulates CCL7 expression in human renal proximal tubular epithelial cells [70]. Oncostatin M is upregulated in the early phases of urinary obstruction.…”