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Article Open Access

Integrative bioinformatics and experimental analysis reveals FRA1 as a key mediator of tubulointerstitial inflammation in lupus nephritis

  • Authors:
    • Wenpeng Ni
    • Jialin He
    • Zhouyu Zeng
    • Jialong Ke
    • Runpei Lin
    • Jianming Peng
    • Kunyi Deng
    • Lijuan Wen
    • Yanhui Chen
    • Chilun Zhang
    • Yanfen Li
    • Guanghong Gu
  • View Affiliations / Copyright

    Affiliations: Clinical Laboratory, Boai Hospital of Zhongshan, Zhongshan, Guangdong 528400, P.R. China, Department of Research and Teaching, Boai Hospital of Zhongshan, Zhongshan, Guangdong 528400, P.R. China, Clinical Laboratory, Zhongshan Torch Development Zone People's Hospital, Zhongshan, Guangdong 528400, P.R. China
    Copyright: © Ni et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 103
    |
    Published online on: January 28, 2026
       https://doi.org/10.3892/mmr.2026.13813
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Abstract

Tubulointerstitial injury is a key driver of lupus nephritis (LN) progression, and dysregulation of the immune microenvironment is a central feature of this process. The molecular mediators of this dysregulation remain incompletely defined. In the present study an integrated bioinformatics and experimental analysis was performed of the Activator Protein 1 (AP‑1) family transcription factor Fos‑related antigen 1 (FRA1) in LN tubulointerstitium. Analysis of gene expression omnibus datasets (GSE113342, GSE200306 and GSE127797) showed that FRA1 was markedly upregulated in the tubulointerstitium of LN samples and that its expression positively correlated with CD8+ T cells, regulatory T cells, monocytes, M1 macrophages and activated mast cells, but negatively correlated with plasma cells, resting CD4+ memory T cells, M0/M2 macrophages, resting dendritic cells and resting mast cells. In vivo experiments revealed that, FRA1 expression was also increased in kidneys from MRL/lpr mice. Furthermore, in vitro, lentiviral overexpression of FRA1 in HK‑2 cells induced robust upregulation of IL‑6, IL‑1β, IL‑8, MCP‑1 and RANTES, whereas FRA1 knockdown selectively decreased IL‑6 and RANTES levels. Together, these results indicate that FRA1 is significantly elevated in the LN tubulointerstitium and may foster a proinflammatory microenvironment by regulating key cytokines. The FRA1/AP‑1 axis therefore represents a potential regulator of renal inflammation in LN and a candidate therapeutic target.

View Figures

Figure 1

Workflow of the study. (A)
Differential expression analysis was performed separately on two
public tubulointerstitial datasets (GSE113342 and GSE200306) and
the intersecting up and downregulated DEGs were identified. (B)
Functional enrichment analysis was applied to the intersected DEGs
to explore enriched biological processes and pathways. (C) Among
upregulated DEGs, receiver operating characteristic curve analysis
prioritized FRA1 as the top target gene, which was then subjected
to downstream correlation and immune-infiltration analyses.
CIBERSORT and correlation analyses were performed in GSE127797 to
evaluate relationships between FRA1 expression, immune-cell
fractions and tubular epithelial-related cytokines. (D) In
vivo validation of FRA1 expression was performed by
immunohistochemistry and western blot analysis in MRL/lpr mouse
kidneys. (E) In vitro functional experiments were carried
out in HK-2 cells with lentiviral FRA1 overexpression and knockdown
to assess effects on cytokine expression. Together, these steps
integrate bioinformatic discovery with in vivo and in
vitro validation to evaluate FRA1 as a candidate mediator of
tubulointerstitial inflammation in lupus nephritis. DEGs,
differentially expressed genes; GO, gene ontology; KEGG, Kyoto
encyclopedia of genes and genomes.

Figure 2

Identification of DEGs in LN
tubulointerstitium. Volcano plots showing DEGs between patients
with LN and healthy controls in (A) GSE113342 and (B) GSE200306.
(C) Venn diagram of DEGs overlapping between GSE113342 and
GSE200306, yielding 15 common genes. Heatmap of the 15 shared DEGs
in (D) GSE113342 and (E) GSE200306. DEGs, differentially expressed
genes; LN, lupus nephritis; Not sig, not significant; up,
upregulated; down, downregulated; sig, significant.

Figure 3

Functional enrichment analysis. (A)
Bubble plot illustrating the GO and KEGG enrichment results for the
15 DEGs. (B) Bar plot of GO enrichment analysis (C) KEGG enrichment
network of the 15 DEGs. DEGs, differentially expressed genes; GO,
gene ontology; KEGG, Kyoto encyclopedia of genes and genomes; Bp,
biological process; MF, molecular function.

Figure 4

ROC curves for predictive performance
of five DEGs upregulated in LN group. Panels (A-E) display the ROC
curves for (A) FRA1 (AUC=0.977), (B) CCL19 (AUC=0.898), (C) STAT1
(AUC=0.849), (D) C1R (AUC=0.781) and (E) MX1 (AUC=0.755),
respectively. ROC, receiver operating characteristic; AUC, area
under the curve; TPR, true positive rate; FPR, false positive rate;
CI, confidence interval.

Figure 5

Association between FRA1 and immune
cell infiltration. CIBERSORT-estimated immune cell proportions in
each sample stratified by FRA1 expression (median split): (A)
Low-Fra1 group; (B) high-Fra1 group. (C) Bar plots comparing the
proportions of immune cell types between high- and low-FRA1 groups.
Cells significantly increased in the high-FRA1 group included
CD8+ T cells, T regulatory cells, monocytes, M1
macrophages and activated mast cells; cells reduced in the
high-FRA1 group included plasma cells, resting CD4+
memory T cells, M0/M2 macrophages, resting dendritic cells and
resting mast cells (mean ± SEM; n=44; group comparisons were
performed using two-sided Wilcoxon rank-sum tests; *P<0.05;
**P<0.01, ***P<0.001). (D) Spearman correlation analyses
showing associations between FRA1 expression and immune cell
fractions, as well as tubular epithelial-related cytokines,
including (E) IL-6, (F) IL-1β, (G) IL-8, (H) MCP-1 (CCL2), (I)
RANTES (CCL5), (J) TGF-β and (K) TNF. All correlation tests are
two-sided Spearman rank tests; *P<0.05, **P<0.01 and
***P<0.001.

Figure 6

FRA1 expression in MRL/lpr mouse
kidneys. Representative H&E staining of kidney sections from
20-week-old (A) MRL/lpr mice and (B) MRL/MPJ controls, showing
tubulointerstitial inflammation and damage in MRL/lpr mice. MRL/lpr
mice exhibit diffuse and extensive tubulointerstitial inflammation
and structural damage. (C-H) FRA1 expression in mouse kidney
sections. (C, E and G) Three independent mice from the MRL/lpr
group, all demonstrating elevated FRA1 levels in the tubular
epithelium. (D, F and H) Three independent mice from the control
group, displaying baseline FRA1 immunoreactivity. (I)
Quantification of FRA1-positive area from IHC images. (J) Western
blot analysis of FRA1 protein levels from MRL/lpr and control mice.
(K) Densitometric quantification of FRA1 bands normalized to
β-actin. Scale bar, 50 µm; Data are plotted as the mean ± SEM; n=3
per group; group comparisons were performed using two-sided Welch's
t-tests; ***P<0.001.

Figure 7

Effect of FRA1 on inflammatory
cytokine expression in HK-2 cells. (A) Representative western blots
of FRA1 and inflammatory cytokines (IL-1β, IL-6, and IL-8) in HK-2
cells 144 h after transduction with FRA1-OE, FRA1-shRNA or their
corresponding controls. (B) Representative western blots of MCP-1,
RANTES, TGF-β and TNF-α in HK-2 cells following the same
transduction conditions. Densitometric semi-quantification of
protein bands normalized to β-actin for: (C) FRA1, (D) IL-1β, (E)
IL-6, (F) IL-8, (G) MCP-1, (H) RANTES, (I) TGF-β and (J) TNF-α.
Data are presented as the mean ± SEM; n=3 per group; comparisons
among subgroups were assessed using the two-sided Kruskal-Wallis
test; *P<0.05, **P<0.01 and ***P<0.001. OE, over
expression; ns, not significant; sh, short hairpin.
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Copy and paste a formatted citation
Spandidos Publications style
Ni W, He J, Zeng Z, Ke J, Lin R, Peng J, Deng K, Wen L, Chen Y, Zhang C, Zhang C, et al: <p>Integrative bioinformatics and experimental analysis reveals FRA1 as a key mediator of tubulointerstitial inflammation in lupus nephritis</p>. Mol Med Rep 33: 103, 2026.
APA
Ni, W., He, J., Zeng, Z., Ke, J., Lin, R., Peng, J. ... Gu, G. (2026). <p>Integrative bioinformatics and experimental analysis reveals FRA1 as a key mediator of tubulointerstitial inflammation in lupus nephritis</p>. Molecular Medicine Reports, 33, 103. https://doi.org/10.3892/mmr.2026.13813
MLA
Ni, W., He, J., Zeng, Z., Ke, J., Lin, R., Peng, J., Deng, K., Wen, L., Chen, Y., Zhang, C., Li, Y., Gu, G."<p>Integrative bioinformatics and experimental analysis reveals FRA1 as a key mediator of tubulointerstitial inflammation in lupus nephritis</p>". Molecular Medicine Reports 33.4 (2026): 103.
Chicago
Ni, W., He, J., Zeng, Z., Ke, J., Lin, R., Peng, J., Deng, K., Wen, L., Chen, Y., Zhang, C., Li, Y., Gu, G."<p>Integrative bioinformatics and experimental analysis reveals FRA1 as a key mediator of tubulointerstitial inflammation in lupus nephritis</p>". Molecular Medicine Reports 33, no. 4 (2026): 103. https://doi.org/10.3892/mmr.2026.13813
Copy and paste a formatted citation
x
Spandidos Publications style
Ni W, He J, Zeng Z, Ke J, Lin R, Peng J, Deng K, Wen L, Chen Y, Zhang C, Zhang C, et al: <p>Integrative bioinformatics and experimental analysis reveals FRA1 as a key mediator of tubulointerstitial inflammation in lupus nephritis</p>. Mol Med Rep 33: 103, 2026.
APA
Ni, W., He, J., Zeng, Z., Ke, J., Lin, R., Peng, J. ... Gu, G. (2026). <p>Integrative bioinformatics and experimental analysis reveals FRA1 as a key mediator of tubulointerstitial inflammation in lupus nephritis</p>. Molecular Medicine Reports, 33, 103. https://doi.org/10.3892/mmr.2026.13813
MLA
Ni, W., He, J., Zeng, Z., Ke, J., Lin, R., Peng, J., Deng, K., Wen, L., Chen, Y., Zhang, C., Li, Y., Gu, G."<p>Integrative bioinformatics and experimental analysis reveals FRA1 as a key mediator of tubulointerstitial inflammation in lupus nephritis</p>". Molecular Medicine Reports 33.4 (2026): 103.
Chicago
Ni, W., He, J., Zeng, Z., Ke, J., Lin, R., Peng, J., Deng, K., Wen, L., Chen, Y., Zhang, C., Li, Y., Gu, G."<p>Integrative bioinformatics and experimental analysis reveals FRA1 as a key mediator of tubulointerstitial inflammation in lupus nephritis</p>". Molecular Medicine Reports 33, no. 4 (2026): 103. https://doi.org/10.3892/mmr.2026.13813
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