Tacrolimus Prevents TWEAK-Induced PLA2R Expression in Cultured Human Podocytes

Abstract

<jats:p>Primary membranous nephropathy is usually caused by antibodies against the podocyte antigen membrane M-type phospholipase A2 receptor (PLA2R). The treatment of membranous nephropathy is not fully satisfactory. The calcineurin inhibitor tacrolimus is used to treat membranous nephropathy, but recurrence upon drug withdrawal is common. TNF superfamily members are key mediators of kidney injury. We have now identified key TNF receptor superfamily members in podocytes and explored the regulation of PLA2R expression and the impact of tacrolimus. Data mining of single cell transcriptomics and glomerular transcriptomics data identified TNFRSF12a/Fn14 as the highest expressed TNF receptor superfamily gene in human membranous nephropathy, and this was confirmed by immunohistochemistry that also identified NFκB activation in membranous nephropathy podocytes. Additionally, glomerular transcriptomics identified PLA2R1 expression as being increased in membranous nephropathy in the parenteral administration of the Fn14 ligand TWEAK increased podocyte PLA2R expression in mice. Furthermore, in cultured human podocytes, TWEAK increased the expression of PLA2R as well as the expression of other genes recently identified by GWAS as linked to membranous nephropathy: NFKB1 and IRF4. Interestingly, IRF4 encodes the FK506-binding protein 52 (FKBP52), a protein associated with tacrolimus. Tacrolimus prevented the increased expression of PLA2R, NFKB1 and IRF4 induced by TWEAK in cultured podocytes. In conclusion, TWEAK upregulates the expression of PLA2R and of other genes linked to membranous nephropathy in podocytes, and this is prevented by tacrolimus. An impact of tacrolimus on the expression of PLA2R and other genes in podocytes may underlie its efficacy in treating the disease as well as the frequent recurrence of nephrotic syndrome upon tacrolimus withdrawal.</jats:p>

Exploring the Differences in Molecular Mechanisms and Key Biomarkers Between Membranous Nephropathy and Lupus Nephritis Using Integrated Bioinformatics Analysis

“…There are also many cases of IMN recurrence after transplantation. Cuarental et al increased PLA2R expression in renal podocytes by injecting TWEAK to 14-week-old C57/ BL mice, but proteinuria was not reported in this study (Cuarental et al, 2020). Moreover, PLA2R is not a protein specifically expressed in the kidney.…”

Lyso-Gb3 Increases αvβ3 Integrin Gene Expression in Cultured Human Podocytes in Fabry Nephropathy

“…Undifferentiated podocytes were maintained at 33 °C in RPMI 1640 with penicillin, streptomycin, ITS (insulin, transferrin, selenite), and 10% FCS. At 70 to 80% confluence in 60-mm culture dishes, the temperature was increased to 37 °C for at least 14 days, until differentiation was confirmed [ 16 , 18 , 19 , 20 , 21 , 22 , 23 , 24 , 25 ]. The differentiation state was routinely assessed by well-known morphological changes corresponding to podocyte differentiation ( Figure 1 ).…”

“…Podocyte cultures that failed to differentiate were discarded. Expression of podocyte markers such as nephrin, synaptopodin, or densin were periodically assessed to ensure correct identification [ 16 , 18 , 19 , 20 , 21 , 22 , 23 , 24 , 25 ]. Cells were cultured in serum-free media from 24 h before adding 100 nM of lyso-Gb3 (Sigma, St. Louis, MO, USA) and throughout the experiment.…”

Recent Clinical Trials Insights into the Treatment of Primary Membranous Nephropathy

“…It is unclear whether the difference relates to the drug used or to patient characteristics. However, on top of immune suppression, TAC decreased podocyte PLA2R expression, which may also contribute to the early immunological response [ 39 ].…”