Javier Perez-Hernandez1, Maria D Olivares1, Maria J Forner2, Felipe J Chaves3, Raquel Cortes1, Josep Redon2. 1. Genotyping and Genetic Diagnosis Unit, INCLIVA Biomedical Research Institute, Avd. Menéndez Pelayo, acceso 4, 46010 Valencia, Spain Cardiometabolic and Renal Unit, INCLIVA Biomedical Research Institute, Avd. Menéndez Pelayo, acceso 4, 46010 Valencia, Spain. 2. Cardiometabolic and Renal Unit, INCLIVA Biomedical Research Institute, Avd. Menéndez Pelayo, acceso 4, 46010 Valencia, Spain CIBER Physiopathology of Obesity and Nutrition (CIBEROBN), Institute of Health Carlos III, Minister of Health, Madrid, Spain. 3. Genotyping and Genetic Diagnosis Unit, INCLIVA Biomedical Research Institute, Avd. Menéndez Pelayo, acceso 4, 46010 Valencia, Spain CIBER of Diabetes and Associated Metabolic Diseases (CIBERDEM), Institute of Health Carlos III, Minister of Health, Barcelona, Spain.
Abstract
BACKGROUND: Currently, renal biopsy remains the gold standard for the diagnosis and prognosis of lupus nephritis (LN). However, it is an invasive method, and new non-invasive laboratory tests are needed to identify renal involvement without renal biopsy. Podocyte damage plays an important role in the pathogenesis and progression of systemic lupus erythematosus (SLE). We characterize whether the phenotype of urinary podocytes (viability, apoptosis, mRNA and protein levels of the podocyte-associated molecules) is a novel marker of clinical and histological features in SLE patients with or without LN. METHODS: We quantified in urinary sediments of 32 SLE patients and 20 controls, mRNA and protein levels of podocalyxin, synapotopodin, podocin, nephrin and WT-1 by quantitative real-time polymerase chain reaction and western blot analysis and correlated these with clinical and histological parameters. The viability of detached urine podocytes was analysed by flow cytometry with podocalyxin and annexin V/7-AAD double staining and immunofluorescence of urine podocyte cultures. RESULTS: The degree of a poptotic podocytes from urine samples was significantly decreased in patients with LN, especially in the active state (33% compared with 75% in controls, P < 0.001), and the majority of the detached podocytes in the urine of patients with active LN were viable (70% grew in culture). Furthermore, urinary mRNA of podocyte-associated molecules was significantly lower in patients with active LN (P < 0.05) compared with healthy controls, and protein levels of podocyte markers were significantly increased in SLE patients, especially with LN compared with SLE without LN (P < 0.05) and the healthy control group (P < 0.01). Finally, urinary protein levels of podocyte-related markers were associated with proteinuria and histological features (P < 0.05 and P < 0.01), and receiver operating characteristics curves of protein levels discriminate between LN and healthy controls with an area under the curve (AUC) between 0.91 and 0.77 (P < 0.001). CONCLUSIONS: Urinary dedifferentiated podocytes were shown in active LN, and their protein levels correlated with proteinuria and histological features in LN. These preliminary results suggest that it could be a potentially useful non-invasive marker for evaluating the progression of glomerular disease in SLE.
BACKGROUND: Currently, renal biopsy remains the gold standard for the diagnosis and prognosis of lupus nephritis (LN). However, it is an invasive method, and new non-invasive laboratory tests are needed to identify renal involvement without renal biopsy. Podocyte damage plays an important role in the pathogenesis and progression of systemic lupus erythematosus (SLE). We characterize whether the phenotype of urinary podocytes (viability, apoptosis, mRNA and protein levels of the podocyte-associated molecules) is a novel marker of clinical and histological features in SLEpatients with or without LN. METHODS: We quantified in urinary sediments of 32 SLEpatients and 20 controls, mRNA and protein levels of podocalyxin, synapotopodin, podocin, nephrin and WT-1 by quantitative real-time polymerase chain reaction and western blot analysis and correlated these with clinical and histological parameters. The viability of detached urine podocytes was analysed by flow cytometry with podocalyxin and annexin V/7-AAD double staining and immunofluorescence of urine podocyte cultures. RESULTS: The degree of a poptotic podocytes from urine samples was significantly decreased in patients with LN, especially in the active state (33% compared with 75% in controls, P < 0.001), and the majority of the detached podocytes in the urine of patients with active LN were viable (70% grew in culture). Furthermore, urinary mRNA of podocyte-associated molecules was significantly lower in patients with active LN (P < 0.05) compared with healthy controls, and protein levels of podocyte markers were significantly increased in SLEpatients, especially with LN compared with SLE without LN (P < 0.05) and the healthy control group (P < 0.01). Finally, urinary protein levels of podocyte-related markers were associated with proteinuria and histological features (P < 0.05 and P < 0.01), and receiver operating characteristics curves of protein levels discriminate between LN and healthy controls with an area under the curve (AUC) between 0.91 and 0.77 (P < 0.001). CONCLUSIONS: Urinary dedifferentiated podocytes were shown in active LN, and their protein levels correlated with proteinuria and histological features in LN. These preliminary results suggest that it could be a potentially useful non-invasive marker for evaluating the progression of glomerular disease in SLE.
Authors: Victor G Puelles; James W van der Wolde; Nicola Wanner; Markus W Scheppach; Luise A Cullen-McEwen; Tillmann Bork; Maja T Lindenmeyer; Lukas Gernhold; Milagros N Wong; Fabian Braun; Clemens D Cohen; Michelle M Kett; Christoph Kuppe; Rafael Kramann; Turgay Saritas; Claudia R van Roeyen; Marcus J Moeller; Leon Tribolet; Richard Rebello; Yu By Sun; Jinhua Li; Gerhard Müller-Newen; Michael D Hughson; Wendy E Hoy; Fermin Person; Thorsten Wiech; Sharon D Ricardo; Peter G Kerr; Kate M Denton; Luc Furic; Tobias B Huber; David J Nikolic-Paterson; John F Bertram Journal: JCI Insight Date: 2019-09-19
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