Elsevier

Clinical Immunology

Volume 145, Issue 2, November 2012, Pages 108-121
Clinical Immunology

Inhibition of the TWEAK/Fn14 pathway attenuates renal disease in nephrotoxic serum nephritis

https://doi.org/10.1016/j.clim.2012.08.008Get rights and content

Abstract

Previously it was shown that the TNF superfamily member TWEAK (TNFSF12) acts through its receptor, Fn14, to promote proinflammatory responses in kidney cells, including the production of MCP-1, RANTES, IP-10 and KC. In addition, the TWEAK/Fn14 pathway promotes mesangial cell proliferation, vascular cell activation, and renal cell death. To study the relevance of the TWEAK/Fn14 pathway in the pathogenesis of antibody-induced nephritis using the mouse model of nephrotoxic serum nephritis (NTN), we induced NTN by passive transfer of rabbit anti-glomerular antibodies into Fn14 knockout (KO) and wild type (WT) mice. Severe proteinuria as well as renal histopathology were induced in WT but not in Fn14 KO mice. Similarly, a pharmacologic approach of anti-TWEAK mAb administration into WT mice in the NTN model significantly ameliorated proteinuria and improved kidney histology. Anti-TWEAK treatment did not affect the generation of mouse anti-rabbit antibodies; however, within the kidney there was a significant decrease in glomerular immunoglobulin deposition, as well as macrophage infiltrates and tubulointerstitial fibrosis. The mechanism of action is most likely due to reductions in downstream targets of TWEAK/Fn14 signaling, including reduced renal expression of MCP-1, VCAM-1, IP-10, RANTES as well as Fn14 itself, and other molecular pathways associated with fibrosis in anti-TWEAK treated mice. Thus, TWEAK/Fn14 interactions are instrumental in the pathogenesis of nephritis in the NTN model, apparently mediating a cascade of pathologic events locally in the kidney rather than by impacting the systemic immune response. Disrupting TWEAK/Fn14 interactions may be an innovative kidney-protective approach for the treatment of lupus nephritis and other antibody-induced renal diseases.

Highlights

ā–ŗ Following pathogenic Ab transfer, nephritis was attenuated in Fn14 deficient mice. ā–ŗ Anti-TWEAK mAb treatment ameliorated proteinuria and improved renal histology. ā–ŗ TWEAK blockade decreased renal inflammation, macrophage infiltration and fibrosis. ā–ŗ Inhibiting TWEAK is a novel approach with a localized mechanism of action.

Introduction

Involvement of the kidney, lupus nephritis (LN), is a major determinant in the prognosis of patients with systemic lupus erythematosus (SLE). Using intensive induction and maintenance regimens, long term outcomes have greatly improved the prognosis of patients with lupus nephritis. However, treatment all too often results in less than a complete response [1]. In addition, the incidence of end stage renal disease may in fact be increasing in certain subpopulations [2], [3], [4]. Therefore, while there have been many important advances in our understanding of the pathogenesis of lupus nephritis, these have yet to translate into significant enough gains in the arena of treatment of human disease. Moreover, most of the current therapies that are employed to treat LN are non-specific, and are associated with major side effects. Thus, a kidney-protective modality that would improve the renal prognosis would be of tremendous benefit to LN patients.

Interactions between members of the TNF-ligand superfamily (i.e. CD40, BLyS/BAFF) and their cognate TNF-receptor superfamily members (CD40L, BAFF-R/TACI/BCMA, respectively) are instrumental in the pathogenesis of SLE. Indeed, many studies have shown that inhibition of signaling transduced by these receptor/ligand pairs is beneficial in animal models of lupus featuring nephritis [5], [6], [7], [8], [9]. Notably, an anti-BLyS mAb has recently achieved approval for clinical treatment of SLE [10]. Thus, targeting other TNF family members is also of interest for the development of novel therapeutic approaches for treatment of SLE.

TWEAK is a distinct member of the TNF-ligand superfamily with relevance to the pathogenesis of LN. TWEAK, produced primarily as a soluble cytokine by tissue-infiltrating leukocytes, promotes NF-ĪŗB and MAPK activation through its sole signaling receptor Fibroblast Growth Factor Inducible 14 (Fn14), a member of the TNF receptor family [11], [12]. Notably, Fn14 is expressed at relatively low levels in normal tissues and is highly induced in injured and diseased tissues, thereby activating the TWEAK/Fn14 pathway locally in those target tissues. Previously, we and others have shown that Fn14 can be expressed by cell types that comprise the kidney, including mesangial cells, podocytes, endothelial cells and tubular cells [13], [14], [15], [16]. Interestingly, TWEAK acts on these cell types to induce proinflammatory cytokines and chemokines, including MCP-1 and RANTES which have been implicated in LN [17], [18], as well as vascular activation, including the upregulation of adhesion molecules which are also relevant to LN [19], [20]. In addition to its proinflammatory activity, TWEAK can promote mesangial cell proliferation [14], [15] and tubular cell death [21], [22]. Finally, in the chronic graft versus host (cGVH) model of induced autoimmunity, inhibition of the TWEAK pathway significantly improved glomerulonephritis [23].

Passive transfer into rodents of heterologous sera containing pre-formed antibodies against the glomerular basement membrane (GBM) induces a rapidly progressive, proliferative crescentic glomerulonephritis (GN), modeling human anti-GBM disease (also known as Goodpasture's syndrome). In this rodent experimental model, heterologous antibody-mediated injury primarily involves linear antibody deposition, complement deposits, and acute neutrophil-mediated glomerular injury, followed by an autologous phase involving the generation of a host response to the heterologous antibodies. In a widely used variation of nephrotoxic serum nephritis, herein referred to as nephrotoxic nephritis (NTN), the autologous phase is accelerated by immunization of rodents with heterologous immunoglobulin prior to the passive transfer of the anti-GBM antibodies, thereby promoting the deposition of immune complexes and macrophage-mediated injury [24]. As the NTN model shares these and many other features with human LN, it is a valuable model for exploring LN pathogenesis [19], [25], [26], [27], [28], [29]. NTN poses a high hurdle for therapeutic intervention, considering the accelerated kinetics of nephritis as compared to spontaneous disease in susceptible mouse strains. However, considering the mode of induction, it offers the ability to assess the contribution of mechanisms acting locally in the disease target end organ [30].

To explore a possible role for the TWEAK/Fn14 pathway in the pathogenesis of nephritis mediated by pathogenic antibodies and the potential utility of anti-TWEAK antibodies as a kidney-protective approach for the treatment of lupus nephritis, we investigated the effects of inhibiting this pathway on the development and progression of NTN in mice.

Section snippets

Mice

Seven to eight week old 129/SvJ (129) mice were purchased from The Jackson Laboratory (Bar Harbor, Maine) and housed 3ā€“5 mice per cage in the animal facility of the Albert Einstein College of Medicine (Bronx, NY). Fn14 +/+ (WT) and āˆ’/āˆ’ (KO) mice on the 129/Svev (129) background were provided by Biogen Idec investigators. Mice were acclimatized in the facility for 2Ā weeks prior to experiments. The housing conditions were controlled, with a temperature of 21ā€“23Ā Ā°C and a 12:12Ā hour light:dark cycle.

Fn14 KO mice are resistant to renal injury from pathogenic anti-GBM antibodies

To determine if the TWEAK/Fn14 pathway promotes the pathogenesis of immune mediated nephritis, we induced nephrotoxic nephritis by passive transfer of pre-formed rabbit anti-mouse glomerular antibodies into Fn14 WT and KO mice pre-immunized with rabbit IgG. Fn14 WT mice exhibited significantly higher proteinuria than Fn14 KO mice on days 7 (2Ā days after transfer of nephrotoxic serum), 14, and 21 (Fig.Ā 1A). When analyzed by grades of proteinuria, also shown in Fig.Ā 1, we found a statistically

Discussion

Kidney involvement, a relatively common clinical manifestation of SLE, remains a major cause of morbidity and mortality in this disease. In this paper, we show that targeting the TWEAK/Fn14 pathway, by Fn14 deficiency or anti-TWEAK mAb administration in WT mice, results in significantly reduced severity of nephrotoxic injury induced by passive transfer of pathogenic rabbit anti-GBM antibodies in mice preimmunized with rabbit IgG. This is reflected by significantly decreased proteinuria and

Conclusions

Our results indicate that the TWEAK/Fn14 pathway plays an important role in the pathogenesis of immune nephritis mediated by pathogenic antibodies. Furthermore, anti-TWEAK mAb, when administered alone, attenuated proteinuria as well as both renal glomerular and tubular damage and tubulointerstitial fibrosis. The mechanism of protection afforded by anti-TWEAK mAb treatment was apparently normalizing multiple downstream targets locally in the kidney, including genes that promote inflammation,

Funding

This work was supported by the National Institutes of Health grants RO1 AR048692 and RO1 DK090319, and a research grant from Biogen Idec (to C.P.).

Conflict of interest statement

Drs. Wu, Michaelson, and Burkly are full time employees of Biogen Idec. These studies received partial support from a research grant from Biogen Idec (to C. Putterman).

The following is the supplementary data related to this article.

Supplementary material.

Acknowledgments

We would like to thank the histopathology facility at Biogen Idec for their assistance in the preparation and staining of kidney sections. We acknowledge Suzanne Szak for her assistance with bioinformatics analysis of the Affymetrix data.

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    YX and SC contributed equally to this paper and should be considered co-first authors.

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