Juvenile-onset lupus nephritis (LN) affects up to 80% of juvenile-onset systemic lupus erythematosus patients (JSLE). As the exact role of human renal glomerular endothelial cells (GEnCs) in LN has not been fully elucidated, the aim of this study was to investigate their involvement in LN. Conditionally immortalised human GEnCs (ciGEnCs) were treated with pro-inflammatory cytokines known to be involved in LN pathogenesis and also with LPS. Secretion and surface expression of pro-inflammatory proteins was quantified via ELISA and flow cytometry. NF-κΒ and STAT-1 activation was investigated via immunofluorescence. Serum samples from JSLE patients and from healthy controls were used to treat ciGEnCs to determine via qRT-PCR potential changes in the mRNA levels of pro-inflammatory genes. Our results identified TNF-α, IL-1β, IL-13, IFN-γ and LPS as robustin vitrostimuli of ciGEnCs. Each of them led to significantly increased production of different pro-inflammatory proteins, including; IL-6, IL-10, MCP-1, sVCAM-1, MIP-1α, IP-10, GM-CSF, M-CSF, TNF-α, IFN-γ, VCAM-1, ICAM-1, PD-L1 and ICOS-L. TNF-α and IL-1β were shown to activate NF-κB, whilst IFN-γ activated STAT-1. JSLE patient serum promoted IL-6 and IL-1β mRNA expression. In conclusion, ourin vitromodel provides evidence that human GEnCs play a pivotal role in LN-associated inflammatory process.

Key Points: Mortality and incidence of AKI do not differ between coronavirus disease 2019 (COVID-19) patients with or without glomerular diseases. The main predictor of AKI is pre-COVID-19 eGFR, independent of the presence of GN. Incomplete kidney function recovery after COVID-19-associated AKI is more common in GN patients than in controls. Background: The acute and long-term effects of severe acute respiratory syndrome coronavirus 2 infection in individuals with GN are still unclear. To address this relevant issue, we created the International Registry of COVID-19 infection in GN. Methods: We collected serial information on kidney-related and -unrelated outcomes from 125 GN patients (63 hospitalized and 62 outpatients) and 83 non-GN hospitalized patients with coronavirus disease 2019 (COVID-19) and a median follow-up period of 6.4 (interquartile range 2.3–9.6) months after diagnosis. We used logistic regression for the analyses of clinical outcomes and linear mixed models for the longitudinal analyses of eGFR. All multiple regression models were adjusted for age, sex, ethnicity, and renin-angiotensin-aldosterone system inhibitor use. Results: After adjustment for pre-COVID-19 eGFR and other confounders, mortality and AKI did not differ between GN patients and controls (adjusted odds ratio for AKI=1.28; 95% confidence interval [CI], 0.46 to 3.60; P=0.64). The main predictor of AKI was pre-COVID-19 eGFR (adjusted odds ratio per 1 SD unit decrease in eGFR=3.04; 95% CI, 1.76 to 5.28; P<0.001). GN patients developing AKI were less likely to recover pre-COVID-19 eGFR compared with controls (adjusted 6-month post-COVID-19 eGFR=0.41; 95% CI, 0.25 to 0.56; times pre-COVID-19 eGFR). Shorter duration of GN diagnosis, higher pre-COVID-19 proteinuria, and diagnosis of focal segmental glomerulosclerosis or minimal change disease were associated with a lower post-COVID-19 eGFR. Conclusions: Pre-COVID-19 eGFR is the main risk factor for AKI regardless of GN diagnosis. However, GN patients are at higher risk of impaired eGFR recovery after COVID-19-associated AKI. These patients (especially those with high baseline proteinuria or a diagnosis of focal segmental glomerulosclerosis or minimal change disease) should be closely monitored not only during the acute phases of COVID-19 but also after its resolution.

Key Points: In a murine model of cationic bovine serum albumin (cBSA)–induced membranous nephropathy (MN), complement regulator decay-accelerating factor is upregulated and restrains complement activation. Studies using genetic deletion or pharmacological antagonism of C3aR indicate that the main effector mechanism of complement activation in cBSA-induced MN is C3a/C3aR signaling. C3a formation and/or C3aR-mediated signaling represent promising targets for hypothesis-driven therapies for MN. Background: Complement activation is believed to play a major pathogenic role in membranous nephropathy (MN), but its effector mechanisms are still unclear. Even less investigated is the role of podocyte-expressed complement regulators, including decay-accelerating factor (DAF) in disease pathophysiology. Methods: We induced MN by serial injections of cationic bovine serum albumin (cBSA) in WT, DAF^−/−, and C3aR^−/− BALB/c mice and measured disease severity (by albuminuria, BUN, serum albumin, and glomerular histologic changes) and signs of complement activation in the glomeruli (immunofluorescence for C1q, C3b, and membrane attack complex). We also treated DAF^−/− mice with cBSA-induced MN with a selective C3aR antagonist and measured the same readouts. Results: cBSA-induced MN was associated with increased glomerular expression of DAF. Genetic deletion of DAF resulted in increased complement activation and higher disease severity than in WT animals. Treating cBSA-injected DAF^−/− mice with a C3aR antagonist reduced disease severity. Similarly, C3aR^−/− animals were protected from cBSA-induced MN, despite IgG deposition in the glomeruli and complement activation. Evidence of C1q and C3b deposition in the glomeruli of these mice suggest that IgG-cBSA immune complex formation in the glomeruli activates complement through the classical pathway. Conclusions: On cBSA-induced injury, podocytes upregulate DAF expression, which restrains complement activation. However, after prolonged injury, complement activation overcomes DAF regulatory effects leading to the formation of soluble anaphylatoxin C3a that, by signaling through C3aR, promotes glomerular injury and cBSA-induced MN disease progression. Considering the growing number of complement targeting therapies, our findings may have major translational effect on the treatment of patients with MN.

Patients with autosomal dominant polycystic kidney disease (ADPKD), a genetic disease due to mutations of the PKD1 or PKD2 gene, show signs of complement activation in the urine and cystic fluid, but their pathogenic role in cystogenesis is unclear. We tested the causal relationship between complement activation and cyst growth using a Pkd1 ^KO renal tubular cell line and newly generated conditional Pkd1 ^–/– C3 ^–/– mice. Pkd1 -deficient tubular cells have increased expression of complement-related genes ( C3 , C5 , CfB , C3ar , and C5ar1 ), while the gene and protein expression of complement regulators DAF, CD59, and Crry is decreased. Pkd1 ^–/– C3 ^–/– mice are unable to fully activate the complement cascade and are characterized by a significantly slower kidney cystogenesis, preserved renal function, and reduced intrarenal inflammation compared with Pkd1 ^–/– C3 ^+/+ controls. Transgenic expression of the cytoplasmic C-terminal tail of Pkd1 in Pkd1 ^KO cells lowered C5ar1 expression, restored Daf levels, and reduced cell proliferation. Consistently, both DAF overexpression and pharmacological inhibition of C5aR1 (but not C3aR) reduced Pkd1 ^KO cell proliferation. In conclusion, the loss of Pkd1 promotes unleashed activation of locally produced complement by downregulating DAF expression in renal tubular cells. Increased C5a formation and C5aR1 activation in tubular cells promotes cyst growth, offering a new therapeutic target.

Key Points: Genetically induced decay-accelerating factor (DAF) overexpression prevents adriamycin (ADR)-induced focal segmental glomerulosclerosis (FSGS) in mice. Pharmacologic inhibition of DAF cleavage reduces complement activation in the glomeruli and albuminuria in murine ADR-induced FSGS. Inhibition of complement activation represents a valuable therapeutic strategy for FSGS and, potentially, other glomerular diseases.