Association of Tumor Necrosis Factor Alpha-Induced Protein 3 Interacting Protein 1 (<Emphasis Type="Italic">TNIP1</Emphasis>) Gene Polymorphism (rs7708392) with Lupus Nephritis in Egyptian Patients

Lupus nephritis (LN) is a major cause of morbidity and mortality in systemic lupus erythematosus (SLE). Previous studies suggest that mutant A20 binding inhibitor of NF-κB 1 (ABIN1) protein encoded by tumor necrosis factor alpha-induced protein 3 interacting protein 1 (TNIP1) gene is associated with LN via NF-κB dysregulation. The aim of the current study was to evaluate the association of TNIP1 gene SNP rs7708392 with SLE and LN in Egyptian patients. 5′ nuclease Allelic discrimination was used to evaluate the frequency of TNIP1 SNP rs7708392 in 53 patients with LN, 57 SLE patients without nephritis and 85 healthy controls. The genotyping analysis revealed that the CC genotype was more frequent in controls than SLE patients, while GC and GG genotypes were more common in SLE patients. Moreover, the GG genotype and the G allele were significantly more prevalent among LN patient than non-LN patients (P?<?0.001). In LN patients, the most common genotype was GG (56.6%), while among the non-LN patients; the CG genotype was the most common (59.6%). Regression analysis demonstrated that SLE patients carrying only one G allele had a 3.4 folds increased risk for LN. Our results suggested that TNIP1 SNP (rs7708392) might be associated with the LN in Egyptian SLE patients. TNIP1 SNP (rs7708392) might be used to identify patients at risk of developing LN, which could help in early detection and treatment before progression to end-stage renal disease, improving patients’ outcome and quality of life.     

Synovial tissues concentrate secreted APRIL

Introduction

TNF-like weak inducer of apoptosis (TWEAK) has been implicated as a mediator of chronic inflammatory processes via prolonged activation of the NF-κB pathway in several tissues, including the kidney. Evidence for the importance of TWEAK in the pathogenesis of lupus nephritis (LN) has been recently introduced. Thus, TWEAK levels may serve as an indication of LN presence and activity.

Methods

Multicenter cohorts of systemic lupus erythematosus (SLE) patients and controls were recruited for cross-sectional and longitudinal analysis of urinary TWEAK (uTWEAK) and/or serum TWEAK (sTWEAK) levels as potential biomarkers of LN. The performance of TWEAK as a biomarker for nephritis was compared with routinely used laboratory tests in lupus patients, including anti-double stranded DNA antibodies and levels of C3 and C4.

Results

uTWEAK levels were significantly higher in LN patients than in non-LN SLE patients and other disease control groups (P = 0.039). Furthermore, uTWEAK was better at distinguishing between LN and non-LN SLE patients than anti-DNA antibodies and complement levels, while high uTWEAK levels predicted LN in SLE patients with an odds ratio of 7.36 (95% confidence interval = 2.25 to 24.07; P = 0.001). uTWEAK levels peaked during LN flares, and were significantly higher during the flare than at 4 and 6 months prior to or following the flare event. A linear mixed-effects model showed a significant association between uTWEAK levels in SLE patients and their disease activity over time (P = 0.008). sTWEAK levels, however, were not found to correlate with the presence of LN or the degree of nephritis activity.

Conclusions

High uTWEAK levels are indicative of LN, as opposed to non-LN SLE and other healthy and disease control populations, and reflect renal disease activity in longitudinal follow-up. Thus, our study further supports a role for TWEAK in the pathogenesis of LN, and provides strong evidence for uTWEAK as a candidate clinical biomarker for LN.     

Urinary TWEAK as a biomarker of lupus nephritis: a multicenter cohort study

Introduction

TNF-like weak inducer of apoptosis (TWEAK) has been implicated as a mediator of chronic inflammatory processes via prolonged activation of the NF-κB pathway in several tissues, including the kidney. Evidence for the importance of TWEAK in the pathogenesis of lupus nephritis (LN) has been recently introduced. Thus, TWEAK levels may serve as an indication of LN presence and activity.

Methods

Multicenter cohorts of systemic lupus erythematosus (SLE) patients and controls were recruited for cross-sectional and longitudinal analysis of urinary TWEAK (uTWEAK) and/or serum TWEAK (sTWEAK) levels as potential biomarkers of LN. The performance of TWEAK as a biomarker for nephritis was compared with routinely used laboratory tests in lupus patients, including anti-double stranded DNA antibodies and levels of C3 and C4.

Results

uTWEAK levels were significantly higher in LN patients than in non-LN SLE patients and other disease control groups (P = 0.039). Furthermore, uTWEAK was better at distinguishing between LN and non-LN SLE patients than anti-DNA antibodies and complement levels, while high uTWEAK levels predicted LN in SLE patients with an odds ratio of 7.36 (95% confidence interval = 2.25 to 24.07; P = 0.001). uTWEAK levels peaked during LN flares, and were significantly higher during the flare than at 4 and 6 months prior to or following the flare event. A linear mixed-effects model showed a significant association between uTWEAK levels in SLE patients and their disease activity over time (P = 0.008). sTWEAK levels, however, were not found to correlate with the presence of LN or the degree of nephritis activity.

Conclusions

High uTWEAK levels are indicative of LN, as opposed to non-LN SLE and other healthy and disease control populations, and reflect renal disease activity in longitudinal follow-up. Thus, our study further supports a role for TWEAK in the pathogenesis of LN, and provides strong evidence for uTWEAK as a candidate clinical biomarker for LN.     

Functional monocyte chemoattractant protein-1 promoter −2518 polymorphism and systemic lupus erythematosus: a meta-analysis

The monocyte chemoattractant protein-1 (MCP-1) promoter −2518 A/G polymorphism has been reported inconsistently to be associated with systemic lupus erythematosus (SLE) and lupus nephritis (LN). The aim of this study was to explore whether the MCP-1 polymorphism confers susceptibility to SLE or LN. We surveyed studies on the MCP-1 polymorphism and SLE or LN identified by MEDLINE or manual searches. Meta-analysis was conducted on the AA genotype (recessive effect), AA, and AG genotypes (dominant effect), AA versus GG (genotype contrast), and on the A allele of MCP-1 in SLE and LN in each ethnic population studied and on all subjects. Ten studies, which included 1,739 SLE patients and 1,680 controls, were included in our meta-analysis. These consisted of two European, six Asian, one Latin American, and one mixed study. We did not find any association between the MCP-1 −2518 A/G polymorphism and SLE in all subjects or in ethnic groups. Meta-analysis also failed to reveal an association between the MCP-1 −2518 A/G polymorphisms and LN. This meta-analysis on 3419 subjects indicates that no association exists between the MCP-1 −2518 A/G polymorphism and SLE or LN.     

Serum Thiols as a Biomarker of Disease Activity in Lupus Nephritis

Lupus Nephritis (LN) develops in more than half of the Systemic Lupus Erythematous (SLE) patients. However, lack of reliable, specific biomarkers for LN hampers clinical management of patients and impedes development of new therapeutics. The goal of this study was to investigate whether oxidative stress biomarkers in patients with SLE is predictive of renal pathology. Serum biochemical and oxidative stress markers were measured in patients with inactive lupus, active lupus with and without nephritis and compared to healthy control group. To assess the predictive performance of biomarkers, Receiver Operating Characteristic (ROC) curves were constructed and cut-offs were used to identify SLE patients with nephritis. We observed an increased oxidative stress response in all SLE patients compared to healthy controls. Among the several biomarkers tested, serum thiols had a significant inverse association with SLE Disease Activity Index (SLEDAI). Interestingly, thiols were able too aptly differentiate between SLE patients with and without renal pathology, and serum thiol levels were not affected by immunosuppressive drug therapy. The decreased thiols in SLE correlated significantly with serum creatinine and serum C3 levels. Further retrospective evaluation using serum creatinine or C3 levels in combination with thiol’s cutoff values from ROC analysis, we could positively predict chronicity of renal pathology in SLE patients. In summary, serum thiols emerge as an inexpensive and reliable indicator of LN, which may not only help in early identification of renal pathology but also aid in the therapeutic management of the disease, in developing countries with resource poor settings.     

Urine levels of HMGB1 in Systemic Lupus Erythematosus patients with and without renal manifestations

Introduction

Lupus nephritis (LN) is a severe and frequent manifestation of systemic lupus erythematosus (SLE). Its pathogenesis has not been fully elucidated but immune complexes are considered to contribute to the inflammatory pathology in LN. High Mobility Group Box 1 (HMGB1) is a nuclear non-histone protein which is secreted from different types of cells during activation and/or cell death and may act as a pro-inflammatory mediator, alone or as part of DNA-containing immune complexes in SLE. Urinary excretion of HMGB1 might reflect renal inflammatory injury. To assess whether urinary HMGB1 reflects renal inflammation we determined serum levels of HMGB1 simultaneously with its urinary levels in SLE patients with and without LN in comparison to healthy controls (HC). We also analyzed urinary HMGB1 levels in relation with clinical and serological disease activity.

Methods

The study population consisted of 69 SLE patients and 17 HC. Twenty-one patients had biopsy proven active LN, 15 patients had a history of LN without current activity, and 33 patients had non-renal SLE. Serum and urine levels of HMGB1 were both measured by western blotting. Clinical and serological parameters were assessed according to routine procedures. In 17 patients with active LN a parallel analysis was performed on the expression of HMGB1 in renal biopsies.

Results

Serum and urinary levels of HMGB1 were significantly increased in patients with active LN compared to patients without active LN and HC. Similarly, renal tissue of active LN patients showed strong expression of HMGB1 at cytoplasmic and extracellular sites suggesting active release of HMGB1. Serum and urinary levels in patients without active LN were also significantly higher compared to HC. Urinary HMGB1 levels correlated with SLEDAI, and showed a negative correlation with complement C3 and C4.

Conclusion

Levels of HMGB1 in urine of SLE patients, in particular in those with active LN, are increased and correlate with SLEDAI scores. Renal tissue of LN patients shows increased release of nuclear HMGB1 compared to control renal tissue. HMGB1, although at lower levels, is, however, also present in the urine of patients without active LN. These data suggest that urinary HMGB1 might reflect both local renal inflammation as well as systemic inflammation.     

Identification of urinary metabolites that distinguish membranous lupus nephritis from proliferative lupus nephritis and focal segmental glomerulosclerosis

Introduction

Systemic lupus erythematosus (SLE or lupus) is a chronic autoimmune disease, and kidney involvement with SLE, a.k.a. lupus nephritis (LN), is a frequent and severe complication of SLE that increases patient morbidity and mortality. About 50% of patients with SLE encounter renal abnormalities which, if left untreated, can lead to end-stage renal disease. Kidney biopsy is considered the criterion standard for diagnosis and staging of LN using the International Society of Nephrology/Renal Pathology Society (ISN/RPS) classification, which was developed to help predict renal outcomes and assist with medical decision-making. However, kidney biopsy-based classification of LN is highly invasive and impractical for real-time monitoring of LN status. Here, nuclear magnetic resonance (NMR) spectroscopy-based metabolic profiling was used to identify urinary metabolites that discriminated between proliferative and pure membranous LN as defined by the ISN/RPS classification, and between LN and primary focal segmental glomerulosclerosis (FSGS).

Methods

Metabolic profiling was conducted using urine samples of patients with proliferative LN without membranous features (Class III/IV; n = 7) or pure membranous LN (Class V; n = 7). Patients with primary FSGS and proteinuria (n = 10) served as disease controls. For each patient, demographic information and clinical data was obtained and a random urine sample collected to measure NMR spectra. Data and sample collection for patients with LN occurred around the time of kidney biopsy. Metabolic profiling analysis was done by visual inspection and principal component analysis.

Results

Urinary citrate levels were 8-fold lower in Class V LN compared to Class III/IV patients, who had normal levels of urinary citrate (P < 0.05). Class III/IV LN patients had > 10-fold lower levels of urinary taurine compared to Class V patients, who had mostly normal levels (P < 0.01). Class V LN patients had normal urinary hippurate levels compared to FSGS patients, who completely lacked urinary hippurate (P < 0.001).

Conclusions

This pilot study indicated differences in urinary metabolites between proliferative LN and pure membranous LN patients, and between LN and FSGS patients. If confirmed in larger studies, these urine metabolites may serve as biomarkers to help discriminate between different classes of LN, and between LN and FSGS.     

Can measuring urinary biomarkers improve the management of lupus nephritis?

Although renal biopsy is the most accurate way of assessing renal inflammation in patients with lupus nephritis (LN), the technique is invasive and cannot be performed frequently. Currently used blood and urine biomarkers have limited utility in monitoring the activity of nephritis. In a previous issue of Arthritis Research and Therapy, Singh and colleagues showed that measuring urinary levels of vascular cell adhesion molecule 1 could be useful in both diagnosing and monitoring LN. These levels are higher in patients with lupus than controls, are higher in lupus patients who have active renal disease compared with those who do not, and correlate significantly with the histological activity index in renal biopsies of patients with LN.Lupus nephritis (LN) is one of the most severe forms of systemic lupus erythematosus (SLE). If not treated adequately, the disease can result in renal failure or death. In its early stages, LN may be almost asymptomatic and picked up only by carrying out routine blood and urine tests. Measurements of urine protein are especially helpful in this regard. In a previous issue of Arthritis Research and Therapy, Singh and colleagues described tests for three different urinary biomarkers in patients with SLE, investigating which is likely to be most helpful for monitoring renal disease activity [1].Treatment of LN was revolutionized by the introduction of combined corticosteroid/cyclophosphamide regimes and has advanced more recently with the replacement of cyclophosphamide by mycophenolate in many cases [2]. Early diagnosis and treatment has a major impact on clinical outcomes in patients with LN [3], but can also cause treatment-related adverse effects. Being able to identify exactly when treatment is needed during the course of the disease and when maintenance treatment needs to be increased is therefore important. The most accurate way to assess nephritis in patients with SLE is by carrying out a renal biopsy. The histological type of nephritis can be defined and the degree of inflammation can be quantified using an activity index [4]. A high activity index signifies active yet reversible disease, whereas the chronicity index shows irreversible damage.Renal biopsy is an invasive procedure, however, which cannot be repeated whenever a flare of renal lupus is suspected. We would therefore like to have biomarkers, measurable in blood or urine, which rise and fall with renal disease activity and are closely associated with the degree of inflammation in the kidney. Blood markers such as anti-double-stranded DNA, anti-nucleosome and anti-α-actinin antibodies have been studied. There is some evidence that increases in these markers are associated with renal disease activity [5], as measured by indices such as blood albumin and urine protein, but very little evidence for their association with renal biopsy scores.Given that the kidney is the main site of inflammation in LN, however, biomarkers in urine may reflect this inflammation more closely than those in the blood. There has been some interest in using urinary neutrophil gelatinase-associated lipocalin as a marker of renal inflammation in SLE. Studies have shown that urinary (but not serum) neutrophil gelatinase-associated lipocalin levels correlate with measures of renal disease activity and that a rise in urinary neutrophil gelatinase-associated lipocalin at one visit predicts flare of nephritis at the next visit [6,7]. These studies were cross-sectional and did not look at the association between urinary neutrophil gelatinase-associated lipocalin and renal histology.The current paper by Singh and colleagues is also a cross-sectional study but includes data on renal histology and compares three different urinary assays; CXCL16, monocyte chemoattractant protein 1 (MCP-1) and vascular cell adhesion molecule 1 (VCAM-1) [1]. Since all three of these molecules can play a role in the recruitment of inflammatory cells to the nephritic kidney, it would be entirely reasonable to predict that their levels would rise in the urine when active renal inflammation is at its height. The authors review both clinical and murine evidence supporting this prediction for each of these markers [8]. In this study they report creatinine-normalized urinary CXCL16, MCP-1 and VCAM-1 levels in 73 patients with SLE, 13 healthy volunteers and 22 patients with other forms of glomerulonephritis.The study results showed some evidence of utility for all three of these urinary markers. All markers were elevated in patients with SLE compared to healthy controls and correlated with the level of proteinuria. However, the comparison with healthy controls is complicated by the fact that 92% of these controls were Asian whereas none of the patients were. Disease activity in the study was assessed using the Systemic Lupus Erythematosus Disease Activity Index and correlated with MCP-1 and VCAM-1 but not CXCL16. When patients were defined as having active or inactive renal disease, on the basis of the renal components of the Systemic Lupus Erythema tosus Disease Activity Index score alone, MCP-1 and VCAM-1 but not CXCL16 distinguished those two groups. However, this result was based on a relatively small number of patients with inactive renal disease (n = 14), probably because there was a low threshold for considering a patient to have active renal disease - being positive for any single criterion of hematuria, pyuria, proteinuria or casts was sufficient. This difficulty in defining active LN based on disease activity measures is common to many papers of this type and is a major reason why the availability of renal biopsy data from the date of the urine sample in 24 of these patients is so important. These histological data provide a direct objective measure of nephritis and particularly support the utility of urinary VCAM-1 as a biomarker, since only VCAM-1 was significantly correlated with the activity index and also differentiated class IV LN from the other histological types.This study thus provides compelling evidence that measuring urinary VCAM-1 could be an important new biomarker in patients with LN, and some evidence supporting urinary CXCL16 and MCP-1. Investigation into whether an index combining the values of all three markers, along the lines of the serum chemokine index reported by Bauer and colleagues [9], would have more potential to distinguish active LN from inactive LN would be interesting. As suggested by the authors, however, the most important thing is to study levels of these urinary markers longitudinally in patients with LN to see whether they predict changes in activity of nephritis in individual patients over time.     

Relationship between anti-dsDNA, anti-nucleosome and anti-alpha-actinin antibodies and markers of renal disease in patients with lupus nephritis: a prospective longitudinal study

Introduction  

Glomerulonephritis is a major cause of morbidity and mortality in patients with systemic lupus erythematosus (SLE). Deposition of autoantibodies in the glomeruli plays a key role in the development of lupus nephritis (LN). Different groups have proposed that either anti-nucleosome antibodies or antibodies that bind the intrinsic renal antigen, α-actinin, are central to the pathogenesis of LN. These theories have been based mainly on cross-sectional studies in patients and on experiments in animal models. No previous longitudinal studies have compared the relationships between levels of these antibodies and markers of renal function. We assessed how well anti-α-actinin, anti-nucleosome and anti-double-stranded DNA (anti-dsDNA) antibodies reflected renal outcome measures in patients with new-onset LN followed for up to 2 years.     

The -2518 A/G polymorphism in the monocyte chemoattractant protein 1 gene is associated with the risk of developing systemic lupus erythematosus in Argentinean patients: a multicenter study

Systemic lupus erythematosus (SLE) is a systemic, autoimmune disorder. Monocyte chemoattractant protein 1 (MCP-1), a chemokine involved in the recruitment and migration of monocytes/macrophages, has been shown to be increased in the plasma of SLE patients. The aim of our study was to evaluate the possible association of the polymorphism -2518 of the MCP-1 gene with the risk of developing SLE, manifesting lupus nephritis (LN) and with other clinical features of SLE in an Argentinean population. A group of 171 SLE patients and 120 control subjects were examined. Genotypic and allelic frequencies of the MCP-1 -2518 A/G polymorphism showed significant differences between the SLE and the control groups (p=0.001 and p=0.01, respectively). However, the polymorphism showed no association with LN or with the other clinical variables studied. Our results suggest that the presence of the MCP-1 -2518 A/G polymorphism might be a risk factor for developing SLE in genetically predisposed individuals, but it does not seem to have a role in the evolution of the disease in the Argentinean population.