P2X7 Receptor Stimulation Is Not Required for Oxalate Crystal-Induced Kidney Injury

Hannah L Luz,Martin Reichel,Robert J Unwin,Kai-Uwe Eckardt,Kerim Mutig,Ana C Najenson,Frederick W K Tam,Louise M Tonner,Felix Knauf

doi:10.1038/s41598-019-56560-2

Abstract

Oxalate crystal-induced renal inflammation is associated with progressive kidney failure due to activation of the NLRP3/CASP-1 inflammasome. It has been suggested previously that purinergic P2X7 receptor signaling is critical for crystal-induced inflammasome activation and renal injury. Therefore, we investigated the role of the P2X7 receptor in response to crystal-induced cytokine release, inflammation, and kidney failure using in vitro and in vivo models. Dendritic cells and macrophages derived from murine bone marrow and human peripheral blood mononucleated cells stimulated with calcium-oxalate crystals, monosodium urate crystals, or ATP lead to the robust release of interleukin-1beta (IL-1ß). Treatment with the P2X7 inhibitor A740003 or the depletion of ATP by apyrase selectively abrogated ATP-induced, but not oxalate and urate crystal-induced IL-1ß release. In line with this finding, dendritic cells derived from bone marrow (BMDCs) from P2X7−/− mice released reduced amounts of IL-1ß following stimulation with ATP, while oxalate and urate crystal-induced IL-1ß release was unaffected. In sharp contrast, BMDCs from Casp1−/− mice exhibited reduced IL-1ß release following either of the three stimulants. In addition, P2X7−/− mice demonstrated similar degrees of crystal deposition, tubular damage and inflammation when compared with WT mice. In line with these findings, increases in plasma creatinine were no different between WT and P2X7−/− mice. In contrast to previous reports, our results indicate that P2X7 receptor is not required for crystal-induced CKD and it is unlikely to be a suitable therapeutic target for crystal-induced progressive kidney disease.

Highlights

The kidney is highly predisposed to crystalopathies, since one of its main functions is to filter, secrete and concentrate substances via urine formation
In a first series of experiments we investigated IL-1ß cytokine release from bone marrow-derived dendritic cells (BMDCs) in response to calcium oxalate crystals, monosodium urate crystals and ATP in vitro
To examine whether our findings are specific to BMDCs or can be elicited in different mononuclear cells, we further investigated bone marrow-derived macrophages (BMDMs)

Summary

Introduction

The kidney is highly predisposed to crystalopathies, since one of its main functions is to filter, secrete and concentrate substances via urine formation. It has been demonstrated that (1) silica[12,18] and uric acid[10,17] activate the inflammasome via P2X7 signaling, (2) treatment with a P2X7 inhibitor reduces IL-1ß release[10,12,18], (3) oxalate crystal-induced ATP release contributes to kidney inflammation[8] and (4) several studies have suggested that P2X7 may be a suitable pharmacological target in various renal diseases, BMDMs and human PBMCs in vitro. Full-length gels are presented in Supplementary Fig. 1 Such as diabetic nephropathy[19,20], glomerulonephritis[21], hypertension[20], kidney injury induced by metabolic syndrome[22] and ischemic acute kidney injury[23]. The present study is directed at defining the role of P2X7 receptor in crystal-induced inflammation and kidney disease

Methods

Results

Conclusion