Crosstalk Between Th17 Cells and Renal Tubular Epithelial Cells Promotes Fibrotic Progression in IgA Nephropathy.

Abstract

Objective: Th17 cell-mediated immune injury is a crucial factor contributing to tubulointerstitial fibrosis in patients with IgA nephropathy (IgAN). However, the exact mechanisms by which Th17 cells induce tubulointerstitial fibrosis remain to be fully elucidated.

Methods: An IgAN mouse model was established and validated. Transcriptome sequencing, combined with bioinformatics analysis, was carried out to explore the immune injury pathways in renal tissues and the activation pathways in Th17 cells that were co-cultured with tubular epithelial cells. In subsequent experiments, small interfering RNA (siRNA) and overexpression plasmids were used to manipulate cellular targets. Validation was conducted through quantitative real-time polymerase chain reaction (qPCR), Western blotting, and immunofluorescence assays.

Results: Compared with the control mice, IgAN mice exhibited elevated serum creatinine levels and increased urine protein-to-creatinine ratios. Renal pathological examination revealed the characteristic features of IgAN. Transcriptomic analysis of the kidney tissues from the model mice showed the activation of Th17 differentiation pathways, which was further confirmed by immunofluorescence analysis showing increased expression of interleukin-17A (IL-17A). These findings indicate an increased abundance of Th17 cells with potential pathogenic significance. When Th17 cells were co-cultured with tubular epithelial cells, the level of interleukin-9 (IL-9) in the system increased. This increase in IL-9 activated the Janus kinase 1-signal transducer and activator of transcription 3 (JAK1-STAT3) pathway through the IL-9 receptor (IL-9R) and upregulated the signature transcription factor retinoic acid-related orphan receptor gamma (ROR-γ), thus promoting Th17 cell differentiation. When IL-9R was silenced using siRNA or when the activity of STAT3 was inhibited, both the levels of phosphorylated STAT3 (p-STAT3) and ROR-γ decreased. Moreover, IL-17A secreted by Th17 cells promoted the nuclear translocation of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) in tubular epithelial cells by activating the IL-17 receptor A (IL-17RA)-adaptor protein Act1-tumor necrosis factor receptor-associated factor 6 (TRAF6) complex. This process regulated the production of inflammatory cytokines and drove the initiation and progression of fibrosis. Treatment with a STAT3 inhibitor in IgAN mice led to a reduction in the number of renal Th17 cells and alleviated the fibrotic phenotype.

Conclusion: This study demonstrated that the interaction between Th17 cells and tubular epithelial cells triggers excessive extracellular matrix deposition in the tubulointerstitium, thereby exacerbating the fibrotic phenotype and accelerating the progression of IgAN.