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AB - MP033PURE MICROSCOPIC HEMATURIA, ESPECIALLY FAMILIAL, IN EARLY LIFE, RESULTING FROM HETEROZYGOUS COL4A3/A4 MUTATIONS AND CFHR5 C3 NEPHRITIS MAY BE RESPONSIBLE FOR MANY CASES OF CHRONIC RENAL FAILURE AND ESKD IN LATER LIFE Yiannis Athanasiou Yiannis Athanasiou 1 Nicosia General Hospital, Nicosia, Cyprus Michalis Zavros Michalis Zavros 1 Nicosia General Hospital, Nicosia, Cyprus Maria Arsali Maria Arsali 2 Limassol General Hospital, Limassol, Cyprus Louiza Papazachariou Louiza Papazachariou 3 University of Cyprus, Nicosia, Cyprus Panayiota Demosthenous Panayiota Demosthenous 3 University of Cyprus, Nicosia, Cyprus Isavella Savva Isavella Savva 3 University of Cyprus, Nicosia, Cyprus Konstantinos Voskarides Konstantinos Voskarides 3 University of Cyprus, Nicosia, Cyprus Constantinos Deltas Constantinos Deltas 3 University of Cyprus, Nicosia, Cyprus Alkis Pierides Alkis Pierides 4 Hippocrateon Hospital, Nicosia, Cyprus Abstract Introduction and Aims: According to classical nephrology practice, pure microscopic hematuria [MH] in early life is not considered a serious problem and up to date diagnostic studies are not always carried out. Family urine studies to confirm or rule out a familial disease should always be carried out early and if positive, molecular genetics should be used to study the whole family and establish the underlying genetic defect. Methods: During the last 30 years we have searched systematically in a homogeneous population of 650,000 people for familial MH and some 120 such families are under study so far. Molecular genetic studies during the last 10 years have led to the correct genetic diagnosis in 57 such families with a) 28 heterozygous COL4A3/A4 mutations and TBMN, b) 23 CFHR5 and C3 nephritis and c) 6 families with Alport syndrome. Results: Our most striking finding refers to the COL4A3/A4 heterozygous mutations that turn out to be the commonest cause of familial MH and have been identified in 249 carriers in 30 families. Some of these families are very big indeed and two of these mutations, COL4A3-G1334E and COL4A3-G871C, are particularly common. Mutation COL4A3-G1334E was found in 15 families and it accounts for 174 patients. C3 nephritis with the CFHR5 Cypriot mutation appears to be the 2nd commonest cause of familial MH with 150 carriers in 23 families. Classical XLAS, COL4A5 Alport was identified in 3 families with 9 affected males and 8 female carriers, while ARAS was also present in 3 additional families affecting 8 patients. Of the 249 patients with COL4A3/A4 heterozygous mutations 33 have reached ESKD and of the 150 CFHR5 patients 21 have reached ESKD. Conclusions: Our results strengthen the great significance of the heterozygous COL4A3/A4 mutations, not really well understood until after 1996, less than 20 years ago. These mutations are common, lead to TBMN and are the commonest cause of familial MH. More importantly however, these mutations lead much later in life to proteinuria, hypertension, CRF and ESKD and long term care is mandatory with prompt attention to the addition of proteinuria that requires urgent treatment. Heterozygous mutations COL4A3/A4 cause twice more patients to reach ESKD compared to classical XLAS and ARAS but fortunately at a much older age. Molecular genetics should be used more widely for an early diagnosis of this entity and more renal attention should be given to these patients to preserve kidney function and avoid ESKD. MP034ANGIOTENSIN RECEPTOR BLOCKERS AND ANGIOTENSIN CONVERTING ENZYME INHIBITORS (ARB/ACE) SLOW THE PROGRESSION OF NEPHROPATHY IN MALES WITH FABRY DISEASE Sandro Feriozzi Sandro Feriozzi 1 Belcolle Hospital, Viterbo, Italy Amandine Perrin Amandine Perrin 2 Shire, Eysins, Switzerland Michael West Michael West 3 Dalhousie University, Halifax, NS, Canada Kathy Nicholls Kathy Nicholls 4 Royal Melbourne Hospital & University of Melbourne, Melbourne, Australia Gere Sunder-Plassmann Gere Sunder-Plassmann 5 Medical University of Vienna, Department of Nephrology and Dialysis, Vienna, Austria Joan Torras Joan Torras 6 Hospital of Bellvitge IDIBELL, Barcelona, Spain Pablo Neumann Pablo Neumann 7 Servicio de Nefrologia, Hospital Italiano de La Plata, La Plata, Argentina Markus Cybulla Markus Cybulla 8 FGM, Center of Internal Medicine, Müllheim, Germany On Behalf Of The Fabry Outcome Survey Renal Working Group On Behalf Of The Fabry Outcome Survey Renal Working Group Abstract Introduction and Aims: In general, enzyme replacement therapy (ERT) in males with Fabry nephropathy is effective in stabilizing the progression of renal damage. However, patients (pts) with significant proteinuria and arterial hypertension have a faster than average renal progression despite ERT. Angiotensin receptor blockers and/or angiotensin converting enzyme inhibitors (ARB/ACE) are known to reduce proteinuria, control blood pressure and slow the progression of nephropathy. In Fabry nephropathy there are few data on the role of ARB/ACE treatment. To investigate the potential beneficial effect of ARB/ACE on progression of Fabry nephropathy, a retrospective investigation was carried out using data collected in the Fabry Outcome Survey (FOS, Shire). Methods: The inclusion criteria for the study were: adult male pts >18 year on ERT for >5 years and with annually reported data of: glomerular filtration rate estimated with CKD-EPI formula (eGFR), proteinuria/24h, arterial blood pressure, and therapies beside ERT. At present, data regarding 609 males on ERT (agalsidase alfa 0.2mg/kg/every other week) are reported in FOS (data extracted November 2012); 121 of them met all criteria. Results: Data regarding 43 pts with hypertension and 78 with normal blood pressure were available. Of those with hypertension, 13 were receiving ARB/ACE (ARB/ACE+) and 30 were treated with other antihypertensive drugs (ARB/ACE-). The table shows eGFR and proteinuria changes. The annualized rates of change in eGFR (CKD-EPI) and total proteinuria between the 3 groups above were not significantly different (p=0.861 for CKD-EPI, p=0.873 for total protein). Interestingly, however, in ARB/ACE+ the yearly slope of eGFR was less than in ARB/ACE- and even lower than in normotensives; these results occurred despite the fact that baseline eGFR was lower in ARB/ACE+ than in the other 2 groups. There was a greater reduction of proteinuria in ARB/ACE+ pts compared with ARB/ACE-. Notably in normotensives, proteinuria increased. In ARB/ACE+ blood pressure mean was: systolic: 138.4±21.3 mmHg at baseline and 120.9±13.4 mmHg at 5 years; diastolic: 85.3±14.1 mmHg at baseline and 77.3±9.3 mmHg at 5 years. Blood pressure in ARB/ACE- was: systolic: 124.6±15.8 mmHg at baseline and 121.1±14.3 mmHg at 5 years; diastolic: 73.0±11.3 mmHg at baseline and 72.6±9.6 mmHg at 5 years. Conclusions: In Fabry nephropathy with ERT, ARB/ACE treatment, compared with other antihypertensives, and with equal BP control, is associated with reductions in proteinuria and in eGFR annual slope that are lower than in normotensive pts. Sample sizes were small, so caution is required; however, our findings suggest a potential clinical benefit from ARB/ACE in stabilizing renal disease in men with Fabry nephropathy treated with agalsidase alfa. Open in new tabDownload slide Open in new tabDownload slide MP035ECULIZUMAB REDUCES TERMINAL COMPLEMENT (TC) AND COMPLEMENT ALTERNATIVE PATHWAY (CAP) ACTIVATION, INFLAMMATION, ENDOTHELIAL DAMAGE, THROMBOSIS AND RENAL INJURY IN ATYPICAL HEMOLYTIC UREMIC SYNDROME (AHUS) PATIENTS Roxanne Cofiell Roxanne Cofiell 1 Alexion Pharmaceuticals, Inc., Cheshire, CT Anjli Kukreja Anjli Kukreja 1 Alexion Pharmaceuticals, Inc., Cheshire, CT Krystin Bedard Krystin Bedard 1 Alexion Pharmaceuticals, Inc., Cheshire, CT Yan Yan Yan Yan 1 Alexion Pharmaceuticals, Inc., Cheshire, CT Angela Mickle Angela Mickle 1 Alexion Pharmaceuticals, Inc., Cheshire, CT Masayo Ogawa Masayo Ogawa 1 Alexion Pharmaceuticals, Inc., Cheshire, CT Camille Bedrosian Camille Bedrosian 1 Alexion Pharmaceuticals, Inc., Cheshire, CT Susan Faas Susan Faas 1 Alexion Pharmaceuticals, Inc., Cheshire, CT Abstract Introduction and Aims: aHUS is a genetic, life-threatening disease of chronic, uncontrolled complement activation and systemic complement-mediated thrombotic microangiopathy (TMA). Eculizumab (Ecu) blocks TC activation, inhibits TMA, normalizes platelets (plts) and improves renal function in aHUS patients (pts). To elucidate mechanisms by which Ecu exerts clinical benefit in aHUS pts, we studied biomarkers of complement and endothelial (EC) activation, inflammation, EC damage, thrombosis and renal injury prior to and during Ecu treatment. Methods: Biomarkers were evaluated in normal healthy volunteers (NHV; N=10-20) and adult aHUS pts (N=26-38) at baseline (BL) and during 1 year of Ecu treatment in a prospective clinical trial. Results: Prior to Ecu (BL), all biomarker levels were significantly elevated in most pts (Table), including pts with prior plasma exchange/infusion (PE/PI) (Figure) or normal plts, LDH, and haptoglobin levels. With sustained Ecu, C5a, sC5b-9 and all renal injury markers rapidly normalized, sTNFR1, thrombomodulin, F1+2 and D-dimer levels significantly decreased by up to 77-99%, and Ba and sVCAM-1 (CAP and EC activation) decreased by 30% and up to 60%, respectively, but remained elevated. Conclusions: aHUS pts who were not receiving Ecu, including those with prior PE/PI or normal clinical laboratory values, show TC and CAP activation, elevated markers of inflammation and coagulation, EC activation and damage, thrombosis and renal injury. Ecu normalized TC markers, potently reduced EC damage markers to near-normal levels, markedly reduced inflammatory and thrombosis markers and eliminated signs of ongoing renal injury; these benefits were sustained with ongoing Ecu. Ecu reduced but did not normalize sVCAM-1 and plasma Ba, reflecting ongoing EC and CAP activation that was not pathogenic in the presence of ongoing Ecu. This further indicates that CAP activation was ongoing in aHUS, that CAP alone is not sufficient to cause thrombosis or organ injury, and that it is critical to maintain chronic Ecu to inhibit TC-mediated tissue injury. These data underscore the chronic complement dysregulation and ongoing risk of systemic TMA and organ damage in pts with aHUS, and the requirement for continued TC blockade with Ecu, even when clinical presentation and clinical laboratory values have improved. Open in new tabDownload slide Open in new tabDownload slide Open in new tabDownload slide Open in new tabDownload slide MP036MATERNAL EFFECTS ON THE CIRCADIAN GENE EXPRESSION IN FETAL KIDNEYS Krisztina Mészáros Krisztina Mészáros 1 Center for Pediatrics and Adolescent Medicine, Heidelberg University, Heidelberg, Germany Linda Pruess Linda Pruess 1 Center for Pediatrics and Adolescent Medicine, Heidelberg University, Heidelberg, Germany Matthias Gondan Matthias Gondan 2 Department of Psychology, University of Copenhagen, Copenhagen, Denmark Eberhard Ritz Eberhard Ritz 3 Department of Internal Medicine, Heidelberg University, Heidelberg, Germany Franz Schaefer Franz Schaefer 1 Center for Pediatrics and Adolescent Medicine, Heidelberg University, Heidelberg, Germany Abstract Introduction and Aims: The circadian rhythm of key kidney functions such as electrolyte and water excretion involving blood pressure control is driven by the clock gene network. In adults, broad adverse effects of the disturbed circadian rhythm are known, whereas potential role of the maternal circadian clock disturbances in prenatal programming of renal function has not been studied yet. We sought to explore the effect of altered light exposure and restricted feeding schedule on the development of circadian gene expression in the fetal kidney. Methods: Pregnant rats were randomly allocated into 5 groups. The rats were fed ab libitum under 12h:12h light-dark cycle (LD), constant light (LL), constant darkness (DD), or ultradian, 6h:6h light-dark cycle (6-6LD). Another group of pregnant rats was kept under 12h:12h light-dark cycle with food access restricted to the light period (FR). Mothers (6-10 per group) with their offspring were sacrificed at 4-h intervals one day before the expected delivery. Intrarenal circadian gene expression patterns were profiled by real-time rtPCR for the canonical clock genes Rev-erbα and Per2 and the clock-controlled genes NHE3, αENaC, SGK1, and AVPR2. Results: Kidneys of fetuses from mothers kept at LD displayed circadian expression of the investigated genes [Rev-erbα (p = 0.014), Per2 (p = 0.014), NHE3 (p < 0.001), αENaC (p < 0.001), SGK1 (p = 0.017), and AVPR2 (p = 0.003)]. Intrarenal gene expression of the fetuses from mothers exposed to LL exhibited robust circadian expression with phases similar to those observed at LD [Rev-erbα (p = 0.027), Per2 (p < 0.001), NHE3 (p < 0.001), αENaC (p< 0.001), SGK1 (p < 0.001), and AVPR2 (p = 0.022)]. In contrast, circadian fluctuations of intrarenal gene expression were completely absent in fetuses with maternal DD and 6-6LDexposure. In the animals exposed to maternal FR circadian rhythmicity persisted for Per2 (p < 0.001), was lost for Rev-erbα and showed a distinct phase shift for the clock-controlled genes [NHE3 (p < 0.001), αENaC (p < 0.001), SGK1 (p = 0.024), and AVPR2 (p = 0.021)]. Conclusions: Our findings show that in the developing kidney the circadian clock network becomes operational even before birth and oscillatory gene expression is affected differently by different types of light cue modification during pregnancy. In addition, the oscillatory expression of kidney specific clock-controlled genes responds to nutritional cues independently of canonical clock gene input. MP037A POLYMORPHISM IN THE MASTER REGULATOR GENE OF THE ANTIOXIDANT SYSTEM (NRF2) PREDICTS INCIDENT CARDIOVASCULAR EVENTS IN STAGE 2-5 CKD PATIENTS. Alessandra Testa Alessandra Testa 1 CNR-IFC, Reggio Calabria, Italy Belinda Spoto Belinda Spoto 1 CNR-IFC, Reggio Calabria, Italy Daniela Leonardis Daniela Leonardis 1 CNR-IFC, Reggio Calabria, Italy Maria Cristina Sanguedolce Maria Cristina Sanguedolce 1 CNR-IFC, Reggio Calabria, Italy Anna Pisano Anna Pisano 1 CNR-IFC, Reggio Calabria, Italy Maria Rosa Parlongo Maria Rosa Parlongo 1 CNR-IFC, Reggio Calabria, Italy Giovanni Tripepi Giovanni Tripepi 1 CNR-IFC, Reggio Calabria, Italy Francesca Mallamaci Francesca Mallamaci 1 CNR-IFC, Reggio Calabria, Italy Carmine Zoccali Carmine Zoccali 1 CNR-IFC, Reggio Calabria, Italy - On Behalf Of The MAURO Working Group - On Behalf Of The MAURO Working Group Abstract Introduction and Aims: The Nuclear factor erythroid-derived 2-like 2 (Nrf2) is a master regulator of genes encoding antioxidant and phase II detoxifying enzymes. Cytoplasmic Nrf2 is bound to a repressor molecule, the Kelch-like ECH-associated protein 1 (Keap-1), which facilitates its proteasome degradation. In the presence of ROS, Nrf2 dissociates from Keap1 and triggers the antioxidant response. Oxidative stress and inflammation are common features in chronic kidney disease (CKD), but the potential involvement of the Nrf2-Keap1 system in cardiovascular disease in CKD patients has not been investigated In this study we tested the hypothesis that an altered Nrf2-Keap1 system may be involved in the high risk for cardiovascular (CV) events in CKD. Methods: We studied 9 SNPs (Single Nucleotide Polymorphism) in Nrf2 gene and 3 SNPs in Keap1 gene characterizing the genetic variability of both the two genes and investigated the relationship between these SNPs and the incidence rate of CV events in a cohort of 759 patients with stage 2-5 CKD. Results: Over a 2.41 years follow-up 124 patients had incident (fatal and non-fatal) CV events. Of the 12 SNPs investigated, the rs11085735 of Keap1 gene was the sole to be associated with CV events. The genotypic distribution of Keap1 rs11085735 SNP did not deviate from Hardy Weinberg Equilibrium (P=0.88). The incidence rate of CV events was twice higher in in patients homozygous (AA) or heterozygous (AC) for the A allele of the rs11085735 SNP (HR: 1.85, 95% CI: 1.20-2.84, P=0.005) than in those with the CC genotype. This relationship was fully confirmed in a multivariate analyses adjusting for a series of traditional risk factors and factors peculiar to CKD (HR: 1.75, 95% CI: 1.13-2.71, P=0.01). Because genetic SNPs are transmitted randomly at mating (Mendelian randomization), most likely such an association is causal in nature. Conclusions: The A allele of the Keap1 rs11085735 SNP is an independent predictor of fatal and non-fatal CV events in patients with CKD of various severity. These data support the hypothesis that polymorphisms in the genes of the Nrf2-Keap1 system represent relevant genetic markers of a high risk for CV events by oxidative stress in this population. MP038DIAGNOSIS OF AUTOSOMAL DOMINANT POLYCYSTIC KIDNEY DISEASE USING EFFICIENT PKD1 AND PKD2 TARGETED NEXT GENERATION SEQUENCING Daniel Trujillano Daniel Trujillano 1 Centre for Genomic Regulation (CRG), Barcelona, Spain Gemma Bullich Gemma Bullich 2 Fundacio Puigvert, Barcelona, Spain Jose Ballarin Jose Ballarin 2 Fundacio Puigvert, Barcelona, Spain Roser Torra Roser Torra 2 Fundacio Puigvert, Barcelona, Spain Xavier Estivill Xavier Estivill 1 Centre for Genomic Regulation (CRG), Barcelona, Spain Elisabet Ars Elisabet Ars 2 Fundacio Puigvert, Barcelona, Spain Abstract Introduction and Aims: Molecular diagnostics of autosomal dominant polycystic kidney disease relies on mutation screening of PKD1 and PKD2, which is complicated by extensive allelic heterogeneity and the presence of six highly homologous sequences of PKD1 (exons 1-33). To date, specific sequencing of the repeated part of PKD1 requires laborious long-range amplifications. The high cost and long turnaround time of PKD1 and PKD2 mutation analysis using conventional techniques limits its widespread application in clinical settings. We developped a new technique for diagnosis of ADPKD based on next generation sequencing. Methods: We performed targeted next generation sequencing of PKD1 and PKD2. Pooled barcoded DNA patient libraries were enriched by in-solution hybridization with PKD1 and PKD2 probes. Bioinformatics analysis was performed using an in-house developed pipeline. We validated the assay in a cohort of 36 patients with previously known PKD1 and PKD2 mutations and five control individuals. Then, we used the same assay and bioinformatics analysis in a discovery cohort of 12 uncharacterized patients. Results: We detected 35 out of 36 known definitely, highly likely, and likely pathogenic mutations in the validation cohort, including two large deletions. In the discovery cohort, we detected 11 different pathogenic mutations in 10 out of 12 patients. Conclusions: Our study demonstrates that laborious long-range PCRs of the repeated PKD1 region can be avoided by in-solution enrichment of PKD1 and PKD2 and next generation sequencing. This strategy significantly reduces the cost and time for simultaneous PKD1 and PKD2 sequence analysis facilitating routine genetic diagnostics of autosomal dominant polycystic kidney disease. MP040ARE URIC ACIDS SERUM LEVELS CAUSALLY INVOLVED IN THE DEVELOPMENT OF CORONARY ARTERY DISEASE, CARDIOVASCULAR MORTALITY OR SUDDEN CARDIAC DEATH? A GENETIC STUDY USING THE MENDELIAN RANDOMISATION APPROACH Marcus E Kleber Marcus E Kleber 1 Heidelberg University, Mannheim, Germany Graciela Delgado Graciela Delgado 1 Heidelberg University, Mannheim, Germany Tanja B Grammer Tanja B Grammer 1 Heidelberg University, Mannheim, Germany Guenther Silbernagel Guenther Silbernagel 2 University of Bern, Bern, Switzerland Bernhard K Kraemer Bernhard K Kraemer 1 Heidelberg University, Mannheim, Germany Winfried Maerz Winfried Maerz 1 Heidelberg University, Mannheim, Germany 3 Synlab Academy, Mannheim, Germany 4 Medical University of Graz, Graz, Austria Abstract Introduction and Aims: Uric acid (UA) is the end product of purine metabolism in humans. Modern lifestyle and the Western diet rich in uric acid raising substances like sugar and especially fructose have led to a dramatic rise in UA serum levels which parallels the increase in lifestyle diseases like obesity, hypertension, diabetes, renal failure, coronary heart disease or stroke. Whether uric acid truly represents an independent risk factor for the development of these diseases is still a matter of debate. An approach to investigate the causality of biomarkers using genetic polymorphisms is Mendelian Randomization. Methods: UA was measured in patients of the Ludwigshafen Risk and Cardiovascular Health Study using a photometric colour test on a Hitachi 717 analyzer (Roche, Mannheim, Germany). We calculated a weighted genetic risk score for UA concentration based on 8 non-pleiotropic UA raising SNPs (GRS8).Continuous variables were compared between groups by univariate analysis of variance (ANOVA). Discrete data is shown as percentages and chi-square test was used for comparisons between groups. Association with disease or mortality was examined by logistic regression analysis and Cox proportional hazard regression analysis, respectively. SPSS 20.0 (IBM SPSS, USA) was used for all analyses. Results: The calculated risk score GRS8 was not associated with any of the investigated biochemical markers except for UA and hsCRP. While there was a significant association of UA with coronary artery disease, peripheral vascular disease, cardiomyopathy, valve disease, atrial fibrillation, hypertension, diabetes and especially chronic kidney disease there was no association of GRS8 with any disease condition.UA was associated with all-cause mortality, cardiovascular mortality (CVM) and sudden cardiac death (SCD), GRS8 was significantly associated with CVM and SCD in fully adjusted models with HR of 1.11 (1.02-1.21) and 1.16 (1.01-1.32) per 1 SD increase of GRS8, respectively. Conclusions: Regarding incident fatal events both serum concentration of UA and the GRS8 were significant predictors of CVM and SCD in Cox regression models adjusted for conventional risk factors. This supports the idea of UA playing a causal role in cardiovascular diseases. Open in new tabDownload slide Open in new tabDownload slide MP041SYNERGY BETWEEN THE PHARMACOLOGICAL CHAPERONE 1-DEOXYGALACTONOJIRIMYCIN AND THE HUMAN RECOMBINANT ALPHA-GALACTOSIDASE A IN CULTURED RENAL TUBULAR CELLS FROM PATIENTS WITH FABRY DISEASE Eleonora Riccio Eleonora Riccio 1 Federico II University, Naples, Italy Antonio Pisani Antonio Pisani 1 Federico II University, Naples, Italy Abstract Introduction and Aims: We recently demonstrated in fibroblast cell lines of Fabry Disease (FD) patients a synergistic effect between Enzyme Replacement Therapy (ERT) and Pharmacological Chaperone Therapy (PCT). However, although easy to obtain and to manipulate in cultures, the fibroblasts are not the preferred target of therapy in FD.Therefore, the aim of our study was to test the combination of recombinant human alpha-galactosidase A (rh-alpha-Gal A) with the chaperone molecule 1-deoxynojirimycin (DGJ) in cultured renal tubular cells of FD patients. Methods: To study the effect of the DGJ on rh-alpha-Gal A efficacy, the tubular cells of 3 FD patients, derived from renal biopsies, were incubated with 5 nmol/l rh-alpha-Gal A for 24 h, in the absence or in the presence of 20 μmol/l DGJ. Untreated cells or cells incubated with DGJ alone were used for comparison. Results: None of the cell lines showed significant increases in alpha-Gal A activity when incubated with the chaperone alone, confirming that the GLA gene mutations of these patients were non-responsive to chaperone therapy. The correction of alpha-Gal A activity in the cells incubated with the recombinant enzyme alone ranged from 267,41 to 314,37 nmoles 4-methylumbelliferone/mg protein/hour, from 9,78 to 15,02-fold compared to untreated cells. When the cells where co-incubated with DGJ and rh-alpha-Gal A a highly improved correction of intracellular activity was observed. Increases in alpha-Gal A activity ranged from 38,43 to 59,18-fold, compared to untreated cells (A). Compared to the effects of rh-alpha-Gal A alone, co-administration of DGJ and rh-alpha-Gal A resulted in better correction (3,26 to 5,08-fold) of intracellular alpha-Gal A activity. A representative western blot analysis of tubular cells from patient 3 (B) showed a large increase in alpha-Gal A protein in cells treated with rh-alpha-Gal A and DGJ, compared to the cells treated with rh-alpha-Gal A alone, consistent with the high levels of alpha-Gal A enzyme activity obtained with the combination therapy.The enhancing effect of DGJ was not directed to the endogenous defective alpha-Gal A. First, the cell lines studied were from patients with mutations non-responsive to the chaperone. Second, we observed an increase in the amount of fluorochrome-labelled rh-alpha-Gal A in the presence of DGJ, compared to cells incubated with fluorescent rh-alpha-Gal A alone (C): by this approach only the fluorescent exogenous enzyme is detectable and variations in the intensity of fluorescence reflect only the effects on the recombinant enzyme. Finally, the enhancing effect of DGJ on rh-alpha-Gal A was abolished in the presence of mannose-6-phosphate (D). Conclusions: Co-administration of DGJ and rh-alpha-Gal A resulted in all tubular cell lines in a substantial increase in the amount of intracellular enzyme compared with cells treated with rh-alpha-Gal A alone, suggesting improved intracellular stability of the enzyme. Moreover, because of none of the cell lines showed significant increases in alpha-Gal A activity when incubated with the chaperone alone, we provided evidence that this effect is directed towards the exogenous recombinant enzyme and not on the endogenous defective enzyme. MP042HEREDITARY SPASTIC QUADRIPARESIS ASSOCIATED WITH PROTEINURIC PROGRESSIVE KIDNEY DISEASE Ahad A Abdalla Ahad A Abdalla 1 University Hospital Limerick, Limerick, Ireland Andrew F Malone Andrew F Malone 2 Duke University Medical Center, Durham, NC Michelle P. Winn Michelle P. Winn 2 Duke University Medical Center, Durham, NC Tim Goodship Tim Goodship 3 Newcastle upon Tyne, Newcastle, United Kingdom Cornelius Cronin Cornelius Cronin 4 University hospital Limerick, Limerick, Ireland Peter J. Conlon Peter J. Conlon 5 Beaumont Hospital, Dublin, Ireland Liam F. Casserly Liam F. Casserly 1 University Hospital Limerick, Limerick, Ireland Abstract Introduction and Aims: In 1990 a familial association of proteinuric kidney disease with spastic quadriparesis was reported in two sisters. Both the proband and her sister presented with advanced chronic kidney disease and spastic quadriparesis in their mid-30s. They progressed to end-stage kidney disease 2 years after presentation. We report a follow-up of this family and describe the phenotype across 3 generations. Methods: We constructed a pedigree of all first degree relatives of the proband and their offspring. In December 2012 all family members were invited to participate in the study. 20 of 28 family members agreed to participate and provided informed consent. They underwent neurological clinical examination, urinalysis and quantitation of proteinuria. Serum creatinine and an eGFR by MDRD were measured if proteinuria was present. Medical, radiological and pathological records were reviewed. Results: The pedigree is presented in Figure 1 and Table 1 describes the clinical details of affected members. Seven members were identified as having a phenotype with subnephrotic range proteinuria, impaired renal function and features of spastic quadriparesis. Renal ultrasonography demonstrated small kidneys in cases with advanced renal disease but no structural abnormalities were present. Conclusions: The family described here is unique. There are seven affected members in three generations with potential autosomal dominant transmission. The phenotype of progressive kidney disease associated with spastic quadriparesis represents a unique disease association. Open in new tabDownload slide Open in new tabDownload slide Table 1 ID . Age at Diagnosis . Renal Status . Proteinuria on Urinalysis . Renal Pathology . Neurological Examination . 9001 18 CKD 3+ NA Spastic Quadriparesis 1002 40 ESKD 3+ NA Spastic Quadriparesis 1001 36 ESKD 3+ Interstitial Fibrosis & Tubular Cystic dilatation Spastic Quadriparesis 0103 31 ESKD 3+ Thrombotic Microangiopathy Normal 0104 32 ESKD 3+ NA Spastic Quadriparesis 1003 42 ESKD 3+ NA Spastic Quadriparesis 0105 30 CKD 3+ Thrombotic Microangiopathy Spastic Quadriparesis ID . Age at Diagnosis . Renal Status . Proteinuria on Urinalysis . Renal Pathology . Neurological Examination . 9001 18 CKD 3+ NA Spastic Quadriparesis 1002 40 ESKD 3+ NA Spastic Quadriparesis 1001 36 ESKD 3+ Interstitial Fibrosis & Tubular Cystic dilatation Spastic Quadriparesis 0103 31 ESKD 3+ Thrombotic Microangiopathy Normal 0104 32 ESKD 3+ NA Spastic Quadriparesis 1003 42 ESKD 3+ NA Spastic Quadriparesis 0105 30 CKD 3+ Thrombotic Microangiopathy Spastic Quadriparesis Table 1 ID . Age at Diagnosis . Renal Status . Proteinuria on Urinalysis . Renal Pathology . Neurological Examination . 9001 18 CKD 3+ NA Spastic Quadriparesis 1002 40 ESKD 3+ NA Spastic Quadriparesis 1001 36 ESKD 3+ Interstitial Fibrosis & Tubular Cystic dilatation Spastic Quadriparesis 0103 31 ESKD 3+ Thrombotic Microangiopathy Normal 0104 32 ESKD 3+ NA Spastic Quadriparesis 1003 42 ESKD 3+ NA Spastic Quadriparesis 0105 30 CKD 3+ Thrombotic Microangiopathy Spastic Quadriparesis ID . Age at Diagnosis . Renal Status . Proteinuria on Urinalysis . Renal Pathology . Neurological Examination . 9001 18 CKD 3+ NA Spastic Quadriparesis 1002 40 ESKD 3+ NA Spastic Quadriparesis 1001 36 ESKD 3+ Interstitial Fibrosis & Tubular Cystic dilatation Spastic Quadriparesis 0103 31 ESKD 3+ Thrombotic Microangiopathy Normal 0104 32 ESKD 3+ NA Spastic Quadriparesis 1003 42 ESKD 3+ NA Spastic Quadriparesis 0105 30 CKD 3+ Thrombotic Microangiopathy Spastic Quadriparesis MP043TRANSCATHETER ARTERIAL EMBOLIZATION WITH EMBOSPHERE FOR LIVER CYSTS IN AUTOSOMAL DOMINANT POLYCYSTIC KIDNEY DISEASE Saori Nishio Saori Nishio 1 Hokkaido University Graduate School of Medicine, Sappro, Japan Yusuke Sakuhara Yusuke Sakuhara 1 Hokkaido University Graduate School of Medicine, Sappro, Japan Naoko Matsuoka Naoko Matsuoka 1 Hokkaido University Graduate School of Medicine, Sappro, Japan Junya Yamamoto Junya Yamamoto 1 Hokkaido University Graduate School of Medicine, Sappro, Japan Daigo Nakazawa Daigo Nakazawa 1 Hokkaido University Graduate School of Medicine, Sappro, Japan Tasuku Nakagakaki Tasuku Nakagakaki 1 Hokkaido University Graduate School of Medicine, Sappro, Japan Daisuke Abo Daisuke Abo 1 Hokkaido University Graduate School of Medicine, Sappro, Japan Sekiya Shibazaki Sekiya Shibazaki 1 Hokkaido University Graduate School of Medicine, Sappro, Japan Tatuya Atsumi Tatuya Atsumi 1 Hokkaido University Graduate School of Medicine, Sappro, Japan Abstract Introduction and Aims: Polycystic liver disease (PLD) is the most common extrarenal manifestation associated with autosomal dominant polycystic kidney disease (ADPKD). Patients with PLD often suffer from abdominal discomfort, dyspepsia, or dyspnea. Transcatheter arterial embolization (TAE) for hepatic artery branches using metallic coil has been reported as an effective and less invasive treatment for PLD. The disadvantages of coils are the need to use many of them before achieving complete obstruction and high cost. Furthermore, it is difficult to re-treat a patient in whom a previous TAE procedure with metallic coils had failed as a result of recanalization.This study aimed to evaluate the technical safety and effectiveness of TAE using Embosphere for enlarged polycystic liver. Methods: Five PLD patients with severe symptom (1 male, 4 females) underwent TAE for hepatic artery branches using Embosphere100-300μm and 300-500μm. One patient had undergone TAE with metallic coils had failed as a result of recanalization. We evaluated change of hepatic volume and intra-hepatic cyst volume by MRI, symptoms by visual analog scale and FACT-Hep health-related QOL scores before TAE and at 3, 6, 12 months after treatment. Results: Total liver volume before hepatic TAE was 7518cm3(range, 3874 to 9915 cm3), representing marked hepatomegaly.TAE was considered technically successful when the target hepatic arteries were fully embolized, as demonstrated by hepatic arterial angiography performed at completion of the procedure. Technical success was achieved in all cases. No major complication related to TAE was found. Common adverse events were fever, epigastric pain, nausea, and vomiting. Two patients improved symptoms significantly one month after TAE. We found hepatic cyst volume reduction. No patient complained of worsening of the symptoms after the procedure. Conclusions: We suggest that TAE using Embosiphere is effective and safe in treating symptomatic polycystic liver in ADPKD patients, even who had treated by TAE using metallic coils. MP044HIGH THROUGHPUT SEQUENCING IN SPORADIC FORMS OF STEROID-RESISTANT NEPHROTIC SYNDROME IN CHILDREN FREQUENTLY IDENTIFIES HETEROGENEOUS GENETIC ALTERATIONS THAT PREDICT RESISTANCE TO IMMUNOSUPPRESSIVE TREATMENTS Benedetta Mazzinghi Benedetta Mazzinghi 1 Medical Genetics, Meyer Children's University Hospital, Florence, Italy Sabrina Giglio Sabrina Giglio 1 Medical Genetics, Meyer Children's University Hospital, Florence, Italy 2 Department of Clinical and Experimental Biomedical Sciences, University of Florence, Florence, Italy Aldesia Provenzano Aldesia Provenzano 2 Department of Clinical and Experimental Biomedical Sciences, University of Florence, Florence, Italy Francesca Becherucci Francesca Becherucci 3 Pediatric Nephrology Unit, Meyer Children's University Hospital, Florence, Italy Giulia Sansavini Giulia Sansavini 3 Pediatric Nephrology Unit, Meyer Children's University Hospital, Florence, Italy Fiammetta Ravaglia Fiammetta Ravaglia 3 Pediatric Nephrology Unit, Meyer Children's University Hospital, Florence, Italy Rosa Maria Roperto Rosa Maria Roperto 3 Pediatric Nephrology Unit, Meyer Children's University Hospital, Florence, Italy Luisa Murer Luisa Murer 4 Department of Pediatrics, University of Padua, Padua, Italy Laura Lasagni Laura Lasagni 5 Excellence Centre for Research, Transfer and High Education for the Development of de Novo Therapies (DENOTHE), University of Florence, Florence, Italy Marco Materassi Marco Materassi 3 Pediatric Nephrology Unit, Meyer Children's University Hospital, Florence, Italy Paola Romagnani Paola Romagnani 2 Department of Clinical and Experimental Biomedical Sciences, University of Florence, Florence, Italy 3 Pediatric Nephrology Unit, Meyer Children's University Hospital, Florence, Italy 5 Excellence Centre for Research, Transfer and High Education for the Development of de Novo Therapies (DENOTHE), University of Florence, Florence, Italy Abstract Introduction and Aims: Nephrotic syndrome (NS) constitutes one of the most common diagnoses in pediatric nephrology. Children with NS are usually treated with corticosteroids before any diagnostic procedure, and approximately 80% of them respond to such a treatment. According to this observation, pediatric NS has been separated into two broad categories, i.e., steroid-sensitive NS (SSNS) and steroid-resistant NS (SRNS), with the possibility that different pathophysiologic mechanisms are responsible for conferring the response to steroids. The SRNS can be related to a genetic cause such as mutations of genes responsible for ensuring the anatomical and/or functional entirety of the podocyte, but genetic tests are usually performed only in patients with a familial history, an early onset of disease or syndromic traits. Indeed, the burden of genetic alterations has never been specifically analyzed in sporadic cases, and to which extend genetic forms influence response to treatments in these patients is unknown. In this study, we try to establish the role of genetic alterations in determining resistance to steroid and immunosuppressive treatments by exploring the frequency of genetically-caused NS among sporadic patients that cannot be clinically distinguished among each other except for a differential response to therapy. Methods: We designed a custom sequencing array to target exons and part of flanking sequences for all known genes responsible for NS, that was subsequently applied through next generation sequencing to a selected cohort of 25 children affected by sporadic SRNS that was depleted by all patients that also exhibited extra-renal symptoms, that had a familial history of renal disease or of consanguinity, and that had congenital NS. As controls, we used 35 patients that exhibited a similar clinical phenotype but were responsive to steroids. We then retrospectively correlate the results of the genetic screening with clinical data and information about the response to steroid and immunosuppressive treatment. Results: This innovative, fast and cost-efficient strategy allowed to establish the existence of potentially causative genetic mutations in 36% of the sporadic patients that did not respond to steroids while, using the same technical approach, no one of 35 additional patients that exhibited a similar clinical phenotype but were responsive to steroids had genetic mutations in the analyzed genes (χ2 14.824; p<0.001).Genetic alterations also predicted lack of response to immunosuppressive agents; while none of the mutated patient responded to such drugs, there was a 76.4% of responsive patients among the not mutated ones.( χ2 11.471; p<0.005). Conclusions: These results suggest that genetic alterations are present in over one third of children affected by sporadic forms of SRNS and predict resistance to immunosuppressive treatments. In these patients, a comprehensive screening using such an array is required for genetic counseling to the family and may help clinical decision making, in a fast and cost-efficient manner. MP045INVESTIGATING THE MOLECULAR BASIS IN 174 CASES OF JEUNE ASPHYXIATING THORACIC DYSTROPHY AND OTHER NON MOTILE CILIOPATHIES Miriam Schmidts Miriam Schmidts 1 Institute of Child Health, London, United Kingdom Sonia Christou Sonia Christou 1 Institute of Child Health, London, United Kingdom Claudio Cortes Claudio Cortes 2 Institute for Molecular Bioscience, St Lucia, Australia Aideen McInerney-Leo Aideen McInerney-Leo 3 Diamantina Institute, Woolloongabba, Australia Hulya Kayserili Hulya Kayserili 4 Medical Faculty, Istanbul, Turkey Andreas Zankl Andreas Zankl 3 Diamantina Institute, Woolloongabba, Australia Scambler Peter Scambler Peter 1 Institute of Child Health, London, United Kingdom Emma Duncan Emma Duncan 3 Diamantina Institute, Woolloongabba, Australia Uk1ok Consortium Uk1ok Consortium Carol Wicking Carol Wicking 2 Institute for Molecular Bioscience, St Lucia, Australia Philip L Beales Philip L Beales 1 Institute of Child Health, London, United Kingdom Hannah Mitchison Hannah Mitchison 1 Institute of Child Health, London, United Kingdom Abstract Introduction and Aims: Mutations in genes encoding for cilia associated proteins lead to complex developmental defects and although individual ciliopathy diseases are very rare, together they represent a significant disease burden. Genetic heterogeneity and phenotypic as well as genetic overlap has hampered molecular diagnosis in the past but Next Generation Sequencing (NGS) techniques offer new tools.Jeune Asphyxiating Thoracic Dystrophy (JATD) is a rare ciliopathy characterised by short ribs and life-threatening pulmonary hypoplasia, rare polydactyly and sometimes renal, retinal or liver involvement. Aim of this study was to elucidate the molecular basis of JATD and other ciliopathies with renal phenptypes. Methods: Whole Exome Sequencing, Next Generation Sequencing, Immunofluorescence, zebrafish antisense morpholino. Results: We investigated 61 JATD and 53 other ciliopathy cases using Whole Exome Sequencing and further 60 JATD cases using a gene panel. We identified the disease causing gene in > 70% of cases revealing new phenotype-genotype associations. For JATD, we identified mutations in several new genes: WDR60 and WDR34 represent previously uncharacterised human cytoplasmic dynein-2 complex intermediate chains and mutations in WDR34 seem to be the second most common cause of JATD. Protein-protein interaction between human WDR34 and dynein-light chain LC8 as well as proteomics data suggest this is a new dynein-2 intermediate chain but could also represent a novel link to cytoplasmic dynein-1. WDR34 might further influence NF-kappa B signalling via inhibition of TAK-1, a pathway not previously associated with cilia. We also found human mutations in a potential novel mammalian cytoplasmic dynein-2 light chain for which a mouse model is currently under investigation. Last, we identified mutations in IFT172 as a new cause of JATD with severe liver involvement. Conclusions: In summary, mutations in IFT-A genes as well as the IFT-B component IFT172 cause significant extraskeletal symptoms while mutations the cytoplasmic dynein-2 genes DYNC2H1 as well as mutations in the IFT-B component IFT80 result in a predominant skeletal JATD phenotype. Additionally, identification of several new human dynein-2 light- and intermediate chains improve our understanding of the mammalian dynein-2 complex composition. MP046KIDNEY DYSFUNCTION AND HEALTHCARE RESOURCE UTILIZATION IN TUBEROUS SCLEROSIS COMPLEX PATIENTS IN THE NETHERLANDS Matthew Magestro Matthew Magestro 1 Novartis Pharmaceuticals Corporation, Florham Park, NJ Francis Vekeman Francis Vekeman 2 Groupe d’Analyse, Ltée, Montréal, QC, Canada Timothy Nichols Timothy Nichols 3 Analysis Group, Inc., Boston, MA Paul Karner Paul Karner 3 Analysis Group, Inc., Boston, MA Mei Sheng Duh Mei Sheng Duh 3 Analysis Group, Inc., Boston, MA Bhavini Srivastava Bhavini Srivastava 1 Novartis Pharmaceuticals Corporation, Florham Park, NJ Sahar Barjesteh Van Waalwijk Van Doorn-Khosrovani Sahar Barjesteh Van Waalwijk Van Doorn-Khosrovani 4 Novartis Pharmaceuticals Corporation, Arnhem, Netherlands Bernard A Zonnenberg Bernard A Zonnenberg 5 University Medical Center Utrecht, Utrecht, Netherlands Abstract Introduction and Aims: Tuberous sclerosis complex (TSC) is a genetic disorder that is associated with epilepsy, impaired cognition and multiple non-malignant tumors throughout the body (e.g., angiomyolipoma [AML], lymphangioleiomyomatosis [LAM]). AMLs are kidney lesions that affect the majority of TSC patients, increase in size over time, present risk of acute hemorrhage, and can lead to progressive chronic kidney disease (CKD). This study describes the extent of renal dysfunction, the potential roles of AML and embolization, and their implications for healthcare resource utilization (HCRU) prior to the use of everolimus. Methods: A retrospective, longitudinal cohort study was conducted using medical chart data from TSC patients primarily treated at a single center in the Netherlands from January 1990 to April 2012. Patients were followed longitudinally and classified into open cohorts based on CKD stage (estimated from serum creatinine levels), AML severity (using a staging scheme based on the number, size, and extent of AMLs assessed through imaging), and whether they underwent renal embolization, measured at baseline and during follow-up. Patient characteristics and HCRU rates per patient per year (PPPY) were assessed and compared across cohorts using rate ratios (RR) estimated by Poisson regressions. Results: 369 TSC patients were included (49% female; median [range] age 31 [9-82], follow-up time 13 years [1 day-22 years]). 60 patients (16%) were at CKD stage 3 or higher at baseline or during follow-up; these patients tended to have larger AMLs (65% of patients at CKD stage 3 had AML size ≥3.5 cm vs. 33% at CKD stage 1) and more LAM (among patients with LAM data, 50% at CKD stage 3 had LAM vs. 12% at CKD stage 1). Other attributes associated with advanced CKD stages included increased age, female sex, and LAM severity.HCRU rates of scans, surgery, kidney surgery, non-surgical procedures (including embolization), specialist visits, and medication for CKD stage 1 patients were 0.619, 0.073, 0.004, 0.198, 1.309, and 1.266 PPPY, respectively.Advanced CKD stages were associated with higher rates of HCRU. Compared with CKD stage 1 patients, CKD stage 3 patients had higher rates of scans (RR=2.4, P<0.001), kidney surgery (RR=4.6, P=0.007), non-surgical procedures (RR=2.8, P<0.001), specialist visits (RR=2.5, P<0.001), and medication (RR=1.2, P<0.001).HCRU rates for scans, kidney surgery, non-surgical procedures, specialist visits, medication, and dialysis were approximately twice or more as high among patients with AMLs ≥3.5cm in diameter compared to those whose largest AMLs were <3.5cm (RR=2.2, 2.2, 2.3, 2.0, 1.7, and 4.2, respectively, P≤0.035 for all). HCRU rates for the same healthcare resources were also higher among patients who underwent renal embolization vs. those who did not (RR=2.4, 3.0, 3.0, 1.9, 1.4, and 2.4, respectively, P≤0.021 for all). Conclusions: Results from this study suggest a strong association between the extent of renal dysfunction, worsened AML and LAM status, and increased HCRU. Impaired kidney function due to AMLs remains a key concern in TSC patients despite preventive embolization. The significant burden in terms of HRCU of TSC and its associated progressing complications highlights the need for alternative TSC treatments. MP047UNEXPECTEDLY HIGH PREVALENCE OF RARE GENETIC DISORDERS IN KIDNEY TRANSPLANT RECIPIENTS WITH AN UNKNOWN CAUSAL NEPHROPATHY Claudio Musetti Claudio Musetti 1 University of Eastern Piedmont “Amedeo Avogadro”, Department of Translational Medicine, Nephrology and Kidney Transplant, Novara, Italy Marco Quaglia Marco Quaglia 1 University of Eastern Piedmont “Amedeo Avogadro”, Department of Translational Medicine, Nephrology and Kidney Transplant, Novara, Italy Gian M. Ghiggeri Gian M. Ghiggeri 2 Division of Nephrology and Laboratory on Pathophysiology of Uremia, Istituto “G. Gaslini” Hospital, Genoa, Italy Giovanni B. Fogazzi Giovanni B. Fogazzi 3 Nephrology, Dialysis and Renal Tranplant Unit, Fondazione IRCCS “Ospedale Maggiore-Policlinico, Mangiagalli e Regina Elena, Milano, Italy Fabio Settanni Fabio Settanni 4 Endocrinology and Metabolic Disorders Unit, University of Torino, Turin, Italy Renzo L. Boldorini Renzo L. Boldorini 5 Department of Medical Sciences, University of Eastern Piedmont, Novara, Italy Elisa Lazzarich Elisa Lazzarich 1 University of Eastern Piedmont “Amedeo Avogadro”, Department of Translational Medicine, Nephrology and Kidney Transplant, Novara, Italy Andrea Airoldi Andrea Airoldi 1 University of Eastern Piedmont “Amedeo Avogadro”, Department of Translational Medicine, Nephrology and Kidney Transplant, Novara, Italy Cristina Izzo Cristina Izzo 1 University of Eastern Piedmont “Amedeo Avogadro”, Department of Translational Medicine, Nephrology and Kidney Transplant, Novara, Italy Mara Giordano Mara Giordano 5 Department of Medical Sciences, University of Eastern Piedmont, Novara, Italy Piero Stratta Piero Stratta 1 University of Eastern Piedmont “Amedeo Avogadro”, Department of Translational Medicine, Nephrology and Kidney Transplant, Novara, Italy Abstract Introduction and Aims: Patients with a rare genetic disease often reach end-stage renal disease (ESRD) without a correct diagnosis of causal nephropathy: therefore they can receive renal transplantation (KTx) and develop unexpected and even severe complications. Aim of the study was to describe the cases of rare genetic disorders diagnosed after KTx over the past 15 years in our Center, in order to draw clinical lessons for the transplant physician. Methods: We retrospectively assessed all patients who had received a diagnosis of a rare genetic disorder after KTx, focusing on their pre-KTx history and post-KTx complications. Results: More than 30% (278/911) of KTx performed at our Center had not been diagnosed with a causal nephropathy: prevalence of rare genetic disorders in this subgroup was 4.32% (12/278). These included: 2,8 dihydroxyadeninuria (2,8 DHA)-disease (n=2), HNF-1B associated nephropathy (n=2), UMOD-related nephropathy (n=5), Fabry disease (n=1), INF2 focal segmental glomerulosclerosis (n=1) and Senior-Loken syndrome (n=1). In all cases signs and symptoms of disease had been present long time before KTx. 2,8 DHA -nephropathy relapsed in both patients causing an acute renal failure and jeopardising the graft. Conclusions: KTx recipients without a diagnosis of causal nephropathy appear to be a very selected population in which rare genetic diseases might in fact be more common than expected. As even a belated diagnosis after KTx can have a significant impact on their graft and patient survival and on other affected family members, the possibility of a genetic disorder should be kept in mind when dealing with KTx recipients without a known causal nephropathy. MP048THE CRUCIAL ROLE OF IRON METABOLISM IN ERYHTROID HYPOPLASIA/PURE RED CELL APLASIA DUE TO THE DEVELOPMENT OF ANTI-EPO ANTIBODIES Patricia Garrido Patricia Garrido 1 IBILI, Faculty of Medicine, University of Coimbra, Coimbra, Portugal João C Fernandes João C Fernandes 1 IBILI, Faculty of Medicine, University of Coimbra, Coimbra, Portugal 2 Institute for Molecular and Cellular Biology, University of Porto, Porto, United Kingdom Sandra Ribeiro Sandra Ribeiro 3 Institute for Molecular and Cellular Biology, University of Porto, Porto, Portugal 4 Faculty of Pharmacy, University of Porto, Porto, Portugal Luís Belo Luís Belo 3 Institute for Molecular and Cellular Biology, University of Porto, Porto, Portugal 4 Faculty of Pharmacy, University of Porto, Porto, Portugal Elísio Costa Costa Elísio Costa Costa 3 Institute for Molecular and Cellular Biology, University of Porto, Porto, Portugal Flávio Reis Flávio Reis 1 IBILI, Faculty of Medicine, University of Coimbra, Coimbra, Portugal Alice Santos-Silva Alice Santos-Silva 3 Institute for Molecular and Cellular Biology, University of Porto, Porto, Portugal Abstract Introduction and Aims: The current therapeutic use of recombinant human erythropoietin (rHuEPO) to correct anemia in chronic renal failure patients has been associated to some cases of pure red cell aplasia (PRCA), due to the development of cross-reactive anti-EPO antibodies. This complication is very rare; however, the use of EPO biogenerics has increased the number of cases in the last years. The mechanisms underlying the break in immune tolerance remains poorly elucidated. Our aim was to clarify how, facing an erythroid hypoplasia/PRCA condition, the depression of marrow erythroid activity affects iron metabolism, by using an animal model. Methods: Male Wistar rats (200-250g) were divided into 3 groups (n=8): control - saline (s.c.); two rHuEPO groups, treated respectively with 50 and 200 IU/Kg bw/week of Recormon® (Roche Pharmaceuticals), during 12 weeks. At starting and along the protocol, blood samples were collected to assess renal function, hematological and biochemical data, and iron metabolism (iron, transferrin and ferritin). At the end of the protocol, kidney, liver and duodenum were collected for RT-qPCR studies. We studied the gene expression of EPO, EPOR, transferrin receptor 2 (TfR2), hepcidin (Hamp), ferroportin (SLC40A1), hemojuvelin (HJV), transferrin (TF), hemochromatosis (Hfe), divalent metal transporter 1 (DMT1), matriptase-2 (TMPRSS6), interleukin-6 (Il-6) and bone morphogenic protein 6 (BMP6), which was normalized to the housekeeping genes beta-actin and glyceraldehyde 3-phosphate dehydrogenase. Results: The hematologic changes and the detection of anti-EPO antibodies confirmed the development of an antibody-mediated eryhtroid hypoplasia model induced by administration of high rHuEPO doses (200 IU/Kg/week). In fact, erythropoietic stimuli fails due to inhibition of EPO by anti-EPO Abs and serum iron increased. Indeed, there was an overexpression of Tf, TfR2, BMP6, HFE, HJV in the liver and, therefore, in hepcidin; matriptase was downregulated, further contributing to overexpression of hepcidin. The increased serum iron, which results from increased absorption and mobilization, is supported by the overexpression of DMT1 and ferroportin in duodenum and liver, respectively. Considering the EPO inhibition through anti-EPO antibodies, and the consequent reduction in erythropoiesis, it seems that serum iron concentration is the key modulator in this type of erythroid hypoplasia. Actually, it has been recently reported that iron chelation therapy promotes an improvement in erythropoiesis in PRCA, though the mechanism whereby this is achieved is still unclear. Conclusions: This study provides new insights on how rHuEPO-induced erythroid hypoplasia affects iron metabolism, which could be useful to find new targets and improve therapeutic strategies against this increasingly common condition.Acknowledgements: FCT (PTDC/SAU-TOX/114253/2009, SFRH/BD/61020/2009 and PEST-C/SAU/UI3282/2011) and COMPETE. MP049TNFα POLYMORPHISMS IN CHILDREN WITH NEPHROTIC SYNDROME Doaa M Youssef Doaa M Youssef 1 Zagazig University, Cairo, Egypt Amal S Alshal Amal S Alshal 2 Zagazig University, Zagazig University, Egypt Khalid Salah Khalid Salah 3 Zagazig University, Zagazig, Egypt Abd Elrahman Rashed Abd Elrahman Rashed 3 Zagazig University, Zagazig, Egypt Abstract Introduction and Aims: Idiopathic nephrotic syndrome (NS) was proposed to be a disorder of T-cell dysfunction and its associated cytokines including tumor necrosis factor-alpha (TNF-α. our aim is To determine the association and clinical significance of tumor necrosis factor alpha gene polymorphism in childhood onset nephrotic syndrome. Methods: The study was designed as a case control one, evaluated 90 patients with idiopathic NS (60 patients (66.6 %) were steroid sensitive while 30 patients were steroid resistant), in addition to age and sex matched healthy controls. NS patients included 60 steroid sensitive patients and 30 steroid resistant patients. All participants were subjected to careful history taking, thorough clinical examination, laboratory investigations and genetic analysis of TNFα-308 Detection of tumour necrosis factor alpha (TNF-α) -308 G/A genetic polymorphism [by PCR-based restriction fragment length polymorphism (RFLP). Results: By comparing patients and controls regarding TNF-308 genotypes and alleles had no statistically significant differences. While in comparison between steroid sensitive, resistant patients and controls regarding TNF-308 genotypes and alleles had shown that steroid sensitive patients had statistically significant higher frequency of GG genotype and lower frequency of GA phenotype when compared with steroid resistant patients. This was the same when comparing steroid resistant patients with controls. Also, it was noted that steroid resistant patients had statistically significant higher frequency of AA phenotypes when compared when compared with steroid sensitive patients or controls. In addition, it was found that steroid resistant patients had statistically significant higher frequency of GA + AA genotypes when compared with steroid sensitive patients or controls. Furthermore, it was noted that steroid resistant patients had statistically significant less frequency of G alleles when compared with steroid sensitive patients or controls. Conclusions: We concluded that; TNF-α revealed a strong association at genotypic level, as well as at allelic level, demonstrating that it may share in steroid response in NS therapy MP050EFFECT OF EVEROLIMUS ON RENAL ANGIOMYOLIPOMA IN PEDIATRIC PATIENTS WITH TUBEROUS SCLEROSIS COMPLEX (TSC) BEING TREATED FOR SUBEPENDYMAL GIANT CELL ASTROCYTOMA (SEGA) J Chris Kingswood J Chris Kingswood 1 Royal Sussex County Hospital, Brighton, United Kingdom Sergiusz Jozwiak Sergiusz Jozwiak 2 The Children’s Memorial Health Institute, Warsaw, Poland Elena Belousova Elena Belousova 3 Moscow Research Institute of Pediatrics and Pediatric Surgery, Moscow, Russian Federation Michael Frost Michael Frost 4 Minnesota Epilepsy Group, St Paul, MN Rachel Kuperman Rachel Kuperman 5 Children’s Hospital & Research Center Oakland, Oakland, CA E Martina Bebin E Martina Bebin 6 University of Alabama School of Medicine, Birmingham, AL Bruce Korf Bruce Korf 7 University of Alabama at Birmingham, Birmingham, AL J Robert Flamini J Robert Flamini 8 Children’s Healthcare of Atlanta, Atlanta, GA Michael H Kohrman Michael H Kohrman 9 University of Chicago Medicine Comer Children’s Hospital, Chicago, IL Steven Sparagana Steven Sparagana 10 Texas Scottish Rite Hospital for Children, Dallas, TX Joyce Y Wu Joyce Y Wu 11 Mattel Children’s Hospital UCLA, Los Angeles, CA Noah Berkowitz Noah Berkowitz 12 Novartis Pharmaceuticals Corporation, East Hanover, NJ Sara Miao Sara Miao 12 Novartis Pharmaceuticals Corporation, East Hanover, NJ Scott Segal Scott Segal 12 Novartis Pharmaceuticals Corporation, East Hanover, NJ Antonia Ridolfi Antonia Ridolfi 13 Novartis Pharmaceuticals S.A.S., Rueil-Malmaison, France John J Bissler John J Bissler 14 Le Bonheur Children’s Hospital, Memphis, TN David Neal Franz David Neal Franz 15 Cincinnati Children’s Hospital Medical Center, Cincinnati, OH Abstract Introduction and Aims: Everolimus, an oral mTOR inhibitor, was assessed in the EXIST-1 trial (NCT00789828) for the treatment of TSC patients with SEGA. Everolimus was superior to placebo for the primary endpoint, SEGA response rate (35% vs 0%; P<0.0001), and yielded a safety profile consistent with previous reports (initial cutoff 2 March 2011). Following positive results for the initial cutoff, placebo patients were offered open-label everolimus in the study extension phase. In total, 111 patients received ≥1 dose of everolimus, and as of 11 Jan 2013, 95 patients continued to receive everolimus. A predefined exploratory endpoint of the trial was renal angiomyolipoma response rate in the subset of patients with ≥1 target angiomyolipoma (lesion longest diameter ≥1.0 cm) at baseline. Here we present the effect of everolimus on angiomyolipoma growth in TSC patients being treated for SEGA who were <18 years of age at the start of everolimus. Methods: Patients were randomized (2:1) to everolimus 4.5 mg/m2/d, titrated to blood trough levels of 5-15 ng/mL based on tolerability, or placebo. Angiomyolipoma response was defined as a reduction in sum of volumes of all target lesions ≥50% relative to baseline, with no new lesions ≥1 cm in diameter, no increase in kidney volume ≥20% from the lowest value obtained for the patient, and no angiomyolipoma-related bleeding of grade ≥2 (CTCAE, version 3.0). Adverse events (AEs) were assessed every visit and were graded using CTCAE, version 3.0. Results: As of 11 Jan 2013, 27/33 patients <18 years of age with target angiomyolipoma at baseline continued to receive everolimus, while 6 patients discontinued everolimus. The most common reasons for discontinuation were AEs (6.1%) and withdrawal of consent (6.1%). The median age was 11.5 years (range 5.4-17.5 years); median duration of everolimus exposure was 28.7 months (range 1.9-40.5 months). The angiomyolipoma response rate as per central review was 75.8% (95% confidence interval 57.7%-88.9%). Serious AEs occurred in 10 patients (30.3%). One grade 1 event of hematuria occurred. AEs that occurred in ≥20% of patients were mouth ulceration (42.4 %), convulsion (39.4%), stomatitis (39.4%), and nasopharyngitis (24.2%). Conclusions: Everolimus was efficacious in patients <18 years of age who had renal angiomyolipoma and were being treated for SEGA associated with TSC. AEs were consistent with the known safety profile of everolimus in TSC. MP051EARLY PRESENTATION OF CYSTIC KIDNEYS IN A FAMILY WITH A HOMOZYGOUS INVS MUTATION Machteld M Oud Machteld M Oud 1 Radboud University Medical Centre, Nijmegen, Netherlands Bregje W Van Bon Bregje W Van Bon 1 Radboud University Medical Centre, Nijmegen, Netherlands Ernie MHF Bongers Ernie MHF Bongers 1 Radboud University Medical Centre, Nijmegen, Netherlands Alexander Hoischen Alexander Hoischen 1 Radboud University Medical Centre, Nijmegen, Netherlands Carlo L Marcelis Carlo L Marcelis 1 Radboud University Medical Centre, Nijmegen, Netherlands Nicole De Leeuw Nicole De Leeuw 1 Radboud University Medical Centre, Nijmegen, Netherlands Suzanne JJ Mol Suzanne JJ Mol 2 Jeroen Bosch Hospital, Den Bosch, Netherlands Geert Mortier Geert Mortier 3 Antwerp University Hospital and University of Antwerp, Edegem, Belgium Nine VAM Knoers Nine VAM Knoers 4 University Medical Centre Utrecht, Utrecht, Netherlands Han G Brunner Han G Brunner 1 Radboud University Medical Centre, Nijmegen, Netherlands Ronald Roepman Ronald Roepman 1 Radboud University Medical Centre, Nijmegen, Netherlands Heleen H Arts Heleen H Arts 1 Radboud University Medical Centre, Nijmegen, Netherlands Abstract Introduction and Aims: Nephronophthisis (NPHP) is an autosomal recessive cystic kidney disease that is the most frequent monogenetic cause of end-stage renal disease in children. Infantile NPHP, often in combination with other features such as situs inversus, is regularly caused by mutations in the INVS gene. INVS encodes the ciliary protein inversin, and mutations cause dysfunction of the primary cilia. The aim is to find the causative genetic defect in two related fetuses with complex renal cystic disease. Methods: Single nucleotide polymorphism array analysis and whole-exome sequencing were applied to identify the causative gene defects. Expression levels of the gene were determined by qPCR in RNA that was extracted from fetal fibroblasts. The ciliary localization of inversin was analyzed through immunocytochemistry in fetal fibroblasts. Results: We detected a previously unreported homozygous nonsense mutation in the INVS gene in a family with two severely affected fetuses with enlarged cystic kidneys. After detection of this phenotype by ultrasonography the pregnancies were terminated. Both pregnancies were complicated by oligohydramnios andhypoplasia of the lungs. Mild skeletal abnormalities, such as shortening and bowing of long bones, were also detected in one of the fetuses. Analysis of the fibroblasts of one of the fetuses revealed a low expression of INVS. The inversin protein that was produced mislocalized in fibroblasts: in control fibroblasts, inversin is present in the ciliary axoneme as well as at the basal body, whereas in the fetus’ fibroblasts inversin was only detected at the basal body. Conclusions: The severe phenotypes of two fetuses with characteristics of infantile NPHP and Potter sequence are caused by a homozygous nonsense mutation in INVS, which causes nonsense-mediated decay. Nonetheless, a small amount of mutant protein still forms and mislocalizes to the basal body. The detected skeletal abnormalities possibly expand the phenotypic spectrum associated with INVS mutations. MP052GENE PANEL SEQUENCING IN NEPHROGENETIC DIAGNOSTICS AND RESEARCH Albertien M. Van Eerde Albertien M. Van Eerde 1 UMC Utrecht, Utrecht, Netherlands Bert Van Der Zwaag Bert Van Der Zwaag 1 UMC Utrecht, Utrecht, Netherlands Marc R. Lilien Marc R. Lilien 1 UMC Utrecht, Utrecht, Netherlands Kirsten Y Renkema Kirsten Y Renkema 1 UMC Utrecht, Utrecht, Netherlands Martin H. De Borst Martin H. De Borst 2 UMC Groningen, Groningen, Netherlands Gijs Van Haaften Gijs Van Haaften 1 UMC Utrecht, Utrecht, Netherlands Rachel H. Giles Rachel H. Giles 1 UMC Utrecht, Utrecht, Netherlands Gerjan J. Navis Gerjan J. Navis 2 UMC Groningen, Groningen, Netherlands Nine V.A.M. Knoers Nine V.A.M. Knoers 1 UMC Utrecht, Utrecht, Netherlands Abstract Introduction and Aims: Targeted Next Generation Sequencing allows large numbers of genes to be sequenced in a single experiment. For genetically heterogeneous disorders this approach greatly facilitates sequencing of multiple known disease genes. In early 2014 this method was implemented for 22 nephrogenetic indications at the Genetic Diagnostic Laboratory of the UMC Utrecht. We also apply this method to improve genetic diagnostics in Young CKD/ESRD patients (Project CKD Y). In the Netherlands, there are nearly 3000 patients with early onset (<30 years) renal replacement therapy (RRT). 25-40% RRT patients never received a primary renal disease (PRD) diagnosis according to the ERA-EDTA registry classification. This number is similar to other national registry data of early onset renal disease patients. It is likely that in these very young patients, the unknown PRD diagnoses represent CAKUT (congenital anomalies of the kidney and urinary tract) and other congenital/inherited diseases. If the PRD diagnosis is unknown, this hampers the classification of the patients, and the genetic counseling of family members. In CKD-Y two large cohorts of early onset renal disease (from REGaTTA and Parelsnoer) will be analyzed, to determine in how many patients likely monogenetic diagnoses can be reached. Methods: We have designed the “RENome” using SureSelect, Agilent technology including the coding and flanking intronic regions of 175 known renal disease genes, and an additional 200 genes involved in hereditary renal disease or urinary tract development. The regions are captured and subsequently sequenced (SOLiD™ 5500XL platform; Applied Biosystems, Inc). Examples of gene panels within the RENome are: nephrolithiasis (22 genes), renal fanconi syndrome (14 genes), congenital anomalies of the kidney and urinary tract (40 genes) and renal cysts (50 genes). For diagnostics, additional Sanger sequencing is performed when required to increase the percentage of genotyped nucleotides within a gene panel to > 98%. This strategy leads to a mutation detection sensitivity similar to Sanger sequencing. For diagnostic purposes, RENome genes that are not part of a specific gene panel can remain unanalyzed. In cases where a molecular diagnosis can’t be reached, despite a high suspicion of a genetic kidney or urinary tract disorder, the complete sequence data of the RENome can be analyzed on research basis.In the first half (2014) of CKD-Y we will perform RENome sequencing in 200 early onset (<30 years) RRT patients from the REGaTTA cohort. Variants detected by RENome sequencing will be validated by Sanger sequencing and for selected variants the pathogenicity will be functionally studied. Results: Initial diagnostic RENome sequencing results will be available by May 2014. Conclusions: Nephrogenetic gene panel sequencing will facilitate genetic diagnostics and research in patients with likely nephrogenetic disorders. RENome sequencing will enable us to determine in how many young RRT patients rare variants in known renal disease genes are likely to have contributed to their renal disease.AMvE is supported by the Dutch Kidney Foundation and Fonds NutsOhraNVK and RHG are supported by the Dutch Kidney Foundation and EURenOmicsMHdB is supported by the Dutch Kidney Foundation & Netherlands Organisation for Scientific Research MP053GENE-GENE INTERACTION IN RENIN-ANGIOTENSIN-ALDOSTERONE SYSTEM CONTRIBUTES TO END-STAGE RENAL DISEASE SUSCEPTIBILITY IN A HAN CHINESE POPULATION Kuo-Cheng Lu Kuo-Cheng Lu 1 Cardinal-Tien Hospital, New Taipei City, Taiwan 2 School of Medicine, Fu-Jen Catholic University, New Taipei City, Taiwan Sui-Lung Su Sui-Lung Su 3 School of Public Health, National Defense Medical Center, Taipei, Taiwan Abstract Introduction and Aims: Chronic kidney disease progression to end-stage renal disease (ESRD) and the development and severity of various complications are indirectly influenced by genetic and epigenetic factors. Several renin-angiotensin-aldosterone system (RAAS) gene polymorphisms are associated with ESRD. However, the influence of genetic interactions among these RAAS genes on ESRD susceptibility remains unknown. In this study, we investigated whether RAAS gene single nucleotide polymorphisms (SNPs) and their interactions were associated with ESRD. Methods: This was a case-control study for 647 ESRD cases and 644 controls. Angiotensinogen (AGT) [M235T (rs699) and T174M (rs4762)], angiotensin II type I receptor (AGTR1) [A1166C (rs5186) and C573T (rs5182)], angiotensin-converting enzyme (ACE) [I/D (rs1799752) and G2350A (rs4343)], and aldosterone synthase (CYP11B2) [C-344T (rs1799998)] were genotyped and compared between cases and controls to identify SNPs associated with ESRD susceptibility. Multifactor dimensionality reduction (MDR) was used to identify gene-gene interactions. Results: Several RAAS genes were associated with ESRD: AGT M235T, ACE I/D, ACE G2350A, and CYP11B2 C-344T. By MDR analysis, a three-locus model (ACE ID/ACE G2350A/CYP11B2 C-344T) of gene-gene interaction was the best for predicting ESRD risk, and its maximum testing accuracy was 56.08% and maximum cross validation consistency was 9/10. ESRD risk was higher with the simultaneous occurrence of ACE I/D DD-ACE G2350A AA. AGT, ACE, and CYP11B2 gene polymorphisms are associated with ESRD. Conclusions: The gene-gene interaction effects of ACE I/D, ACE G2350A, and CYP11B2 C-344T polymorphisms are more important than individual factors for ESRD development among Han Chinese. MP054MUTATIONS IN CYP24A1 GENE AND IDIOPATHIC INFANTILE HYPERCALCEMIA IN ITALIAN PATIENTS Maddalena Gigante Maddalena Gigante 1 University of Foggia, Foggia, Italy Luisa Santangelo Luisa Santangelo 2 University Hospital “Policlinico Consorziale - Giovanni XXIII”, Bari, Italy Sterpeta Diella Sterpeta Diella 1 University of Foggia, Foggia, Italy Lucia Argentiero Lucia Argentiero 3 University “Aldo Moro”, Bari, Italy Francesca Cianciotta Francesca Cianciotta 3 University “Aldo Moro”, Bari, Italy Marida Martino Marida Martino 3 University “Aldo Moro”, Bari, Italy Elena Ranieri Elena Ranieri 1 University of Foggia, Foggia, Italy Giuseppe Grandaliano Giuseppe Grandaliano 1 University of Foggia, Foggia, Italy Mario Giordano Mario Giordano 2 University Hospital “Policlinico Consorziale - Giovanni XXIII”, Bari, Italy Loreto Gesualdo Loreto Gesualdo 3 University “Aldo Moro”, Bari, Italy Abstract Introduction and Aims: Loss-of-function mutations in the CYP24A1 gene, which encodes the vitamin D-24 hydroxylase, have recently been recognized as a cause of elevated 1,25-dihydroxyvitamin D concentrations, hypercalcemia, hypercalciuria, nephrocalcinosis/nephrolithiasis in infants and adults. However, true prevalence and natural history of this condition are still to be defined. Although, a case report describing two adult patients has been reported in Italian population, to date a comprehensive molecular study of Idiopathic Infantile Hypercalcemia is still lacking. Here, we report our mutation analysis of CYP24A1 gene in 22 Italian patients. Methods: We studied a cohort of 22 pediatric and adult Italian patients with hypercalcemia, suppressed PTH and nephrocalcinosis from 20 families . Data on clinical symptoms and biochemical measures at the time of disease manifestation were collected retrospectively from medical charts. Patients were evaluated during follow-up and biochemical data were obtained with routine methods. Genomic DNA was extracted from peripheral blood leucocytes using standard methods and whole coding sequence of CYP24A1 gene was analyzed in all patients and family members by PCR and direct sequencing. The potential pathogenicity of the newly identified missense mutations was evaluated by combining different in silico methods: PolyPhen and Sorting Intolerant from Tolerant (SIFT). All genetic studies were approved by the ethics committee of Foggia and Bari Universities. Patients or their parents provided written informed consent. Results: Mutations in CYP24A1 gene were found in six analyzed patients (6/22) with a detection rate of 27%. Four out of these patients were compound heterozygotes for their specific mutations, one was homozygote and one presented only one mutation in a single allele. Overall four CYP24A1 mutations, including small deletions and missense mutations, were identified in four unrelated patients and in two siblings. Two out of these mutations were not previously described and were predicted to be non tolerated change affecting protein function with a score of 0.00 by the SIFT program (http://blocks.fhcrc.org/sift/SIFT.html) and to be “probably damaging” with a score of 1 by the PolyPhen program (www.bork.embl-heidelberg.de/PolyPhen). Conclusions: To the best of our knowledge, this is the first report of a CYP24A1 molecular analysis performed in Italian patients. Our study supports a recessive mode of inheritance in five out six patients and confirms that CYP24A1 plays a causal role in some but not all cases of Idiopathic Infantile Hypercalcemia, suggesting that genetic diagnosis could be helpful in a subset of these patients. This study represents a significant advance in the molecular diagnosis of Idiopathic Infantile Hypercalcemia in Italian population because it (1) analyzes the largest Italian cohort with hypercalcemia, hypercalciuria, nephrocalcinosis/nephrolithiasis to date; (2) expands the spectrum of known CYP24A1 pathogenic mutations. MP055TRANSCRIPTOR FACTORS RESPONSIBLE FOR EPO GENE REGULATION, TRIGGERED BY ADMINISTRATION OF RHUEPO HIGH DOSES João Fernandes João Fernandes 1 IBILI, Faculty of Medicine, University of Coimbra, Porto, Portugal 2 Institute for Molecular and Cellular Biology, University of Porto, Porto, Portugal Sandra Ribeiro Sandra Ribeiro 2 Institute for Molecular and Cellular Biology, University of Porto, Porto, Portugal 3 Faculty of Pharmacy, University of Porto, Porto, Portugal Patricia Garrido Patricia Garrido 4 IBILI, Faculty of Medicine, University of Coimbra, Coimbra, Portugal José Sereno José Sereno 4 IBILI, Faculty of Medicine, University of Coimbra, Coimbra, Portugal Elísio Costa Elísio Costa 2 Institute for Molecular and Cellular Biology, University of Porto, Porto, Portugal 3 Faculty of Pharmacy, University of Porto, Porto, Portugal Flávio Reis Flávio Reis 4 IBILI, Faculty of Medicine, University of Coimbra, Coimbra, Portugal Alice Santos-Silva Alice Santos-Silva 2 Institute for Molecular and Cellular Biology, University of Porto, Porto, Portugal 3 Faculty of Pharmacy, University of Porto, Porto, Portugal Abstract Introduction and Aims: Recombinant human erythropoietin (rHuEPO) therapy in hemodialysis patients corrects anemia. There is a marked variability in rHuEPO sensitivity (up to10-fold variability in dose requirements to correct anemia) and about 5-10% of patients show a marked resistance to rHuEPO therapy. The mechanism underlying the development of rHuEPO resistance remains poorly clarified. Since EPO and EPO receptors are expressed in non-renal tissues, the therapeutic induction of EPO expression pathways in those tissues could be a key step in the treatment of anemia. However, this could only be achieved after the elucidation of the cellular/molecular mechanisms that controls non-renal EPO expression. Our aim was to clarify the erythropoietin gene regulation and expression in renal and liver tissue, using an animal model treated high rHuEPO doses, as used in case of resistance to rHuEPO therapy. Methods: Male Wistar rats, 12 weeks old, were divided in 2 groups: rHuEPO (treated 6 weeks with 400 IU/Kg/week), and the control (administrated with saline solution). Hematological and biochemical data were evaluated at starting, middle and at the end of the protocol. Analysis on the expression of several mRNA-encoding transcriptor factors (namely: hypoxia inducible factors - HIF-1α, -2α and -3α; aryl hydrocarbon receptor nuclear translocators - HIF-1β, HIF-2β and HIF-3β; prolyl hydroxylase 2 - PHD2; factor inhibiting HIF - FIH; and, GATA binding proteins among others) involving EPO regulation was also performed in both kidney and liver. Furthermore, EPO concentration (in serum, in liver and kidney) was also determined. Results: The rats treated with rHuEPO presented along the entire protocol a continuous increase of hemoglobin, red blood cells and reticulocyte counts (P < 0.05 for all); the endogenous EPO levels decreased, resulting from a significant down regulation of EPO gene, observed in both liver and kidney. The rHuEPO group didn’t show any significant difference in what concerns to HIF-2α expression, however a significant down-regulation of HIF-1α was seen at kidney level. Moreover, HIF-3α also showed a strong down-regulation in the live. while FIH and PHD2 were significantly up-regulated in both organs. Conclusions: Our results suggest that the transcriptor factors responsible of EPO gene modulation seem to differ in the case of high EPO concentrations, but also between different tissues (liver vs kidney). This study provides new insights on EPO expression pathway, which could be of great value in the future for therapeutic induction of EPO expression (or repression), in the treatment of anaemia or clinical states associated with aberrant EPO gene expression.Acknowledgements: FCT (PTDC/SAU-TOX/114253/2009, SFRH/BD/61020/2009 and PEST-C/SAU/UI3282/2011) and COMPETE. MP056THE PRESENCE OF PLASMID-MEDIATED ESBLS IN UROPATHOGENS ISOLATED FROM PATIENTS WITH CHRONIC PYELONEPHRITIES Olga Chub Olga Chub 1 Kharkiv Medical Academy for Postgraduate Education, Kharkiv, Ukraine Abstract Introduction and Aims: Resistance to beta-lactams has been increasing in the treatment of urinary tract infections, particularly of chronic pyelonephritis. The production of plasmid-mediated extended-spectrum beta-lactamases (ESBLs), that hydrolyze extended spectrum β-lactams, is the major cause of resistance to these drugs. The aim of this study was to detect and determine the genes encoding the ESBLs including bla(TEM), bla(SHV), and bla(CTX-M) among uropathogens, isolated from patients with chronic pyelonephrities. Methods: Forty-eight strains were isolated from patients with chronic pyelonephritis. The commonest isolated organism was Escherichia coli (24/48, 50%). Susceptibility of isolates to 15 different antimicrobial agents was determined using agar disk diffusion method. ESBLs were determined by polymerase chain reaction (PCR) and characterize by direct sequencing of PCR products. Results: Among 48 isolates, 14 (29.3 %) were ESBL producers. Of the 24 E. coli isolates, the beta-lactamase genes were bla(TEM) (2/24, 8.3%), bla(SHV) (2/24, 8.3%), bla(CTX-M) (3/24, 12.5%). Two K. pneumoniae isolates and two E. faecalis isolates were positive for bla(CTX-M) 40% (2/5) and 22.2% (2/9), respectively. P. mirabilis strain carried bla(TEM) and bla(CTX-M). S. aureus strain harboured bla(CTX-M). Of the 14 ESBL producing isolates, 2 (14.3%) coharboured at least two different bla genes. CTX-M ESBLs were the most common. Conclusions: Was determined the prevalence of plasmid-mediated ESBLs in patients with chronic pyelonephritis, who were treated in one of the hospitals in Kharkov, Ukraine. The presence of resistance genes among uropathogens indicates a high incidence between bacteria all over the world due to plasmids. MP057DETERMINATION OF THE RENAL THRESHOLD FOR GLUCOSE EXCRETION IN FAMILIAL RENAL GLUCOSURIC PATIENTS Ines Aires Ines Aires 1 Hospital Curry Cabral, Lisbon, Portugal 2 Nova Medical School, Lisbon, Portugal David Polidori David Polidori 3 Janssen Research and Development, San Diego, CA Ana Rita Santos Ana Rita Santos 1 Hospital Curry Cabral, Lisbon, Portugal Ana Brito Costa Ana Brito Costa 1 Hospital Curry Cabral, Lisbon, Portugal Carlos Simoes Carlos Simoes 1 Hospital Curry Cabral, Lisbon, Portugal José Rueff José Rueff 2 Nova Medical School, Lisbon, Portugal Fernando Nolasco Fernando Nolasco 1 Hospital Curry Cabral, Lisbon, Portugal Joaquim Calado Joaquim Calado 1 Hospital Curry Cabral, Lisbon, Portugal 2 Nova Medical School, Lisbon, Portugal Abstract Introduction and Aims: The relationship between plasma glucose and renal glucose filtration, reabsorption and excretion is usually described as a threshold type. Recently, a simple method to estimate the renal threshold for glucose excretion (RTG), using serial blood glucose sampling and urinary glucose collection in a standardized Mixed Meal Tolerance Test (MMTT) was validated for SGLT2 pharmacological inhibition in patients with diabetes mellitus type 2 (T2DM).We have utilized this method to determine RTG in a small cohort of Familial Renal Glucosuria (FRG) patients. Methods: Five female patients (aged 31 to 61 years) were included in the study. One was diabetic and in 4 we were able to identify SGLT2 mutations (including the T2DM patient). Plasma glucose and urinary glucose excretion were measured and used to calculate RTG. Results: Twenty-four hour urinary glucose excretion (24hUGE) ranged from 0.95 to 70.4 g/1.73m2/d. All patients displayed an abnormally low RTG, the lowest being 43 mg/dl (patient with an 24hUGE of 70.4 g/1.73m2/d) and the highest 124.6 mg/dl (for the diabetic patient with an 24hUGE of 26 g/1.73m2/d). There was no correlation between 24hUGE and RTG in these subjects, due to the amount of 24hUGE depending on plasma glucose and glomerular filtration rate in addition to renal reabsorptive capacity. Conclusions: Determining RTG provides a better characterization of the pathogenicity of mutations and, as such, serves a more appropriate measure for the study of genotype/phenotype correlations in FRG. MP058CONSENSUS RECOMMENDATION ON FABRY DISEASE DIAGNOSIS IN ADULT PATIENTS WITH KIDNEY DISEASE Linda Van Der Tol Linda Van Der Tol 1 Academic Medical Center, Amsterdam, Netherlands Marieke Biegstraaten Marieke Biegstraaten 1 Academic Medical Center, Amsterdam, Netherlands Sandrine Florquin Sandrine Florquin 1 Academic Medical Center, Amsterdam, Netherlands Liffert Vogt Liffert Vogt 1 Academic Medical Center, Amsterdam, Netherlands Marius A Van Den Bergh Weerman Marius A Van Den Bergh Weerman 1 Academic Medical Center, Amsterdam, Netherlands Carla EM Hollak Carla EM Hollak 1 Academic Medical Center, Amsterdam, Netherlands Derralynn A Hughes Derralynn A Hughes 2 Royal Free & University College Medical School, London, United Kingdom Robin H Lachmann Robin H Lachmann 3 National Hospital for Neurology and Neurosurgery, London, United Kingdom João P Oliveira João P Oliveira 4 Hospital São João, Porto, Portugal Alberto Ortiz Alberto Ortiz 5 IIS-Fundacion Jimenez Diaz, Madrid, Spain Einar Svarstad Einar Svarstad 6 Haukeland University Hospital, Bergen, Norway Wim Terryn Wim Terryn 7 Ghent University Hospital, Ghent, Belgium Camilla Tøndel Camilla Tøndel 6 Haukeland University Hospital, Bergen, Norway Stephen Waldek Stephen Waldek 8 Independant Medical Consultant, Manchester, United Kingdom Christoph Wanner Christoph Wanner 9 University of Würzburg, Würzburg, Germany Michael L West Michael L West 10 Dalhousie University, Halifax, NS, Canada Gabor E Linthorst Gabor E Linthorst 1 Academic Medical Center, Amsterdam, Netherlands Abstract Introduction and Aims: Fabry disease (FD) is an X-linked lysosomal storage disorder, characterized by angiokeratoma, neuropathic pain, cornea verticillata, an- or hypohidrosis, and in males by (near) absent α-galactosidase A (AGAL-A) enzyme activity and very high (lyso)globotriaosylceramide (Gb3). Kidneys, heart and brain are often involved, especially in males where chronic kidney disease may start at early adulthood. Screening for FD in patients with kidney disease reveals a large number of individuals with a mutation in the α-galactosidase A (GLA) gene. Characteristic FD features are often absent; the mutation is a genetic variant of unknown significance. These individuals possibly do not suffer from FD and have an uncertain diagnosis of FD. The societal impact of a wrong diagnosis is huge, as it causes inappropriate counseling and initiation of extremely expensive and burdensome enzyme therapy. We present a diagnostic algorithm on how to approach an individual with kidney disease and an uncertain diagnosis of FD. Methods: A systematic review was performed to identify imaging and laboratory criteria that could confirm or exclude FD. A modified Delphi procedure was conducted among 11 FD experts. We aimed to reach consensus on pre-selected criteria from the literature and criteria that were added by the experts. Results: The international guideline on chronic kidney disease was adopted to define chronic kidney disease. Full agreement was reached on the current gold standard, defined as characteristic storage on electron microscopy (EM) in a kidney biopsy, in the absence of medication that may induce similar storage. The criteria ‘renal cysts’ and ‘small kidneys’ on imaging, as well as the ‘Maltese Cross sign’, ‘immunohistochemical staining of Gb3’, ‘high Gb3’ (in the range of classical males) and ‘high protein excretion’ in urine were rejected because of low or uncertain specificity. Since data suggest that Gb3 may be increased in other diseases, its specificity was debated. There was no agreement that this criterion can confirm FD, although 36% of the panelists indicated that ‘high urine Gb3’, in the context of a GLA mutation, is sufficient. In addition, the ‘Maltese Cross sign’ and ‘high urine Gb3’ were selected as red flags, indicating that its presence makes FD more likely, but further assessments are needed. Further research was recommended to gain insight in the specificity of several items and ‘high Gb3’ in particular. Conclusions: In adults with kidney disease, with a GLA mutation and an uncertain diagnosis of FD, a kidney biopsy with EM analysis should be performed to confirm or reject the diagnosis of FD. Other criteria currently cannot substitute for a biopsy in these cases. MP059VITAMIN K ACCUMULATES IN LDL−/− AND APOE−/− MICE Nadine Kaesler Nadine Kaesler 1 University Hospital of the RWTH Aachen, Aachen, Germany Vincent Brandenburg Vincent Brandenburg 1 University Hospital of the RWTH Aachen, Aachen, Germany Elke Theuwissen Elke Theuwissen 2 Maastricht University, Maastricht, Netherlands Cees Vermeer Cees Vermeer 2 Maastricht University, Maastricht, Netherlands Jürgen Floege Jürgen Floege 1 University Hospital of the RWTH Aachen, Aachen, Germany Georg Schlieper Georg Schlieper 1 University Hospital of the RWTH Aachen, Aachen, Germany Thilo Krüger Thilo Krüger 1 University Hospital of the RWTH Aachen, Aachen, Germany Abstract Introduction and Aims: Vitamin K plays a pivotal role in inhibition of vascular calcification via activation of MGP, a potent inhibitor of calcifying arteriosclerosis. After absorption, vitamin K can be found in all lipoprotein fractions; vitamin K1 is mainly found in chylomicron remnants and VLDL whereas vitamin K2 is mainly transported by LDL. Interaction of apolipoprotein E (ApoE) with the LDL receptor mediates cellular uptake. We investigated the cellular uptake of vitamin K in mice deficient for ApoE (ApoE-/-) or the LDL receptor (LDL-R-/-). Methods: Male, 8 week old wildtype, ApoE-/- and LDL-R-/- mice were fed a diet supplemented with 5 g/kg vitamin K1 and 500 mg/kg vitamin K2 (MK4) for 4 weeks. After sacrifice, contents of vitamin K1 and MK4 were determined by isocratic reversed-phase HPLC in liver, kidney, aorta and pooled serum. Carboxylated and uncarboxylated osteocalcin and t-ucMGP were measured in serum by ELISA. Results: After 4 weeks of the vitamin K diet, the vitamin K1 content in liver from wildtype mice was not different from that of ApoE-/- mice (median 10.2 µg/g tissue). In LDL-R-/- mice the hepatic vitamin K1 content increased markedly (60.9 ± 16.8 ng/g). The liver MK4 content was not affected by the genetic background. LDL-R-/- mice also had significantly higher vitamin K1 concentrations in kidney compared to wildtype. Notably, in the aortic wall the vitamin K tissue content increased 9-fold (K1) and 170-fold (MK4) in ApoE-/- mice (K1: 36.6 ± 12.4 vs 4.0 ± 2.1; MK4: 2600 ± 330 vs 125.7 ± 70.3 ng/g). The increase in aortic vitamin K2 content in LDLR-/- was 7-fold. Functional activity of high dose vitamin K supplementation was evidenced by carboxylation (i.e. activation) of osteocalcin in sera of both knockout strains. No significant effects were observed for t-ucMGP, a marker for vascular calcification. Conclusions: Our data demonstrate tissue-specific uptake of vitamin K isoforms (with K1 preferentially accumulating in liver and K2 in extrahepatic tissues). In contrast to current theories, this uptake is independent of ApoE or the LDLR. Open in new tabDownload slide Open in new tabDownload slide MP060GENOTYPE (A) OF ENOS GENE AND R229Q MUTATION OF NPHS2 APPEARS TO BE ASSOCIATED WITH A WORSE OUTCOME IN PATIENTS WITH IGA NEPHROPATHY Dimitris Xydakis Dimitris Xydakis 1 Venizeleio Hospital, Herakleio, Greece George Goulielmos George Goulielmos 2 University of Crete, Heraklion, Greece Ergini Antonaki Ergini Antonaki 3 Venizeleio Hospital, Heraklio, Greece Kostas Stylianoy Kostas Stylianoy 4 University Hospital, Heraklio, Greece Marita Sfakianaki Marita Sfakianaki 5 Internal Medicine, Heraklio, Greece Apostolos Papadogiannakis Apostolos Papadogiannakis 3 Venizeleio Hospital, Heraklio, Greece Eugene Dafnis Eugene Dafnis 6 University of Crete, Heraklio, Greece Abstract Introduction and Aims: IgA Nephropathy, the most common glomerulonephritis throughout the world, is characterized by an IgA1 deposit of in the glomerular mesangium and it has a very variable clinical course from asymptomatic microscopic hematuria till ESRD. It can be attributed to genetic modifiers that increase the risk of progression of the disease. The NO role in glomerular microcirculation and function of mesangial cells may be altered by endothelial NO synthesis gene polymorphism. Several studies have shown the influence of eNOS gene polymorphism on the progression of kidney disease. Podocin, a membrane protein is located exclusively at the podocyte slit diaphragm, part of the glomerular filtration barrier and is encoded by NPHS2 on chromosome 1q25- 31.The aim of this study is to investigate the NPHS2 R229Q mutation and eNOS gene intron 4a/b polymorphism possible association with clinical presentation and progression of IgA nephropathy in a define geographical region (the island of Crete - 0.6 million population) with the same genetic background. Methods: 103 patients with biopsy-proven IgA glomerulonephritis were included in our study and a median follow up period of 5.1±2.8 years. All subjects where genotyped for the intron 4a/b polymorphism and screened by sequencing for NPHS2-R229Q mutation. The end-points were doubling in baseline serum creatinine and/or relapse of proteinuria and/or initiation of dialysis. Results: At baseline, M/F 68/35, age was 41.5±13.5 years, creatinine 1.21±0.74 mg/dl, proteinuria was 1094±1201 mg/24H, 78% of the patients had a baseline creatinine of ≤ 1.3 mg/dl, 57% had a proteinuria of <1000mg/24H. During follow up time, 12 patients (11%) have doubled serum creatinine and 7 of them (8%) started dialysis in a period of 5.1±4.2 years.40 (39%) patients presented eNOS intron 4 polymorphism a (6 aa,34 ab) and 63 (64%) polymorphism b. Patients reaching the outcome exhibited higher systolic blood pressure (144±20 vs 130±20 mmHg, p=0.02), diastolic blood pressure (82,1±13 vs 72,3±15mmHg, p=0.028), higher baseline creatinine (2.18±2,7 vs 1.34±0,8 mg/dl, p=0.009) and more extended tubulointertitial fibrosis (21% vs 13% , p=0.1)15 (14%) patients presented R229Q mutation and it was more common in patients with proteinuria ≥ 500 mg/24H.Combination of polymorphism a (eNOS) and R229Q (NPHS2) was related to an unfavorable outcome, compared with other combinations (HR 3.9 - 95% Cl 1.63-12.5) to 7.2 (95% Cl 2.3-24.4 ) after adjusting for proteinuria, blood pressure and pathological carateristics.In a multivariate Cox proportional hazard analysis, the presence of a allele of eNOS gene, showed an 8-fold hazard to reach the outcome (HR=8.09, p=0.04) after adjusting for age, sex, presence of hypertension, proteinuria and ACE inhibitors treatment. ACE inhibitors treatment was associated with a hazard ratio of five times less (HR=0.17, p=0,05) in reaching the outcome. Conclusions: In our population, the genotype (a) of eNOS gene and R229Q mutation of NPHS2 appears to be associated with a worse outcome. Their interaction in a patient, is a negative prognostic factor for IgA nephropathy. Additionally, The early detection of (a) allele could indicate those patients at increased risk of renal impairment but more extensive studies need to be done. MP061PRIMARY HYPEROXALURIA TYPE I: RESULTS OF A TUNISIAN COHORT Saoussen Mdimegh Saoussen Mdimegh 1 Sahloul University Hospital, Sousse, Tunisia Ibtihel Ben Hadj Mbarek - Fredj Ibtihel Ben Hadj Mbarek - Fredj 1 Sahloul University Hospital, Sousse, Tunisia Amira Moussa Amira Moussa 1 Sahloul University Hospital, Sousse, Tunisia Asma Omezzine Asma Omezzine 1 Sahloul University Hospital, Sousse, Tunisia Dorsaf Zellama Dorsaf Zellama 2 Nephrology Departement, Sahloul University Hospital, Sousse, Tunisia Sameh Mabrouk Sameh Mabrouk 3 Pediatric Department, Sahloul University Hospital, Sousse, Tunisia Noura Zouari Noura Zouari 3 Pediatric Department, Sahloul University Hospital, Sousse, Tunisia Saida Hassayoun Saida Hassayoun 3 Pediatric Department, Sahloul University Hospital, Sousse, Tunisia Jalel Chemli Jalel Chemli 3 Pediatric Department, Sahloul University Hospital, Sousse, Tunisia Abdelattif Achour Abdelattif Achour 2 Nephrology Departement, Sahloul University Hospital, Sousse, Tunisia Ali Bouslama Ali Bouslama 1 Sahloul University Hospital, Sousse, Tunisia Saoussen Abroug Saoussen Abroug 3 Pediatric Department, Sahloul University Hospital, Sousse, Tunisia Abstract Introduction and Aims: Primary hyperoxaluria type I (PH1) is an autosomal recessive inborn error of glyoxlate metabolism, characterized by the accumulation of renal oxalate and often resulting in end-stage renal disease (ESRD). Rare in Europe, It accounts for 13, 5% of cases of ESRD in children Tunisian. The aim of this study was to describe the clinical presentation, molecular analysis and evolution of HP1 in a Tunisian cohort. Methods: The study includes a cohort of 133 patients (71 infantile form and 62 adult form) suspected suffering from PH 1, from 126 unrelated families. Ten of them were diagnosed after family screening. The genetic analysis was made by PCR/RFLP to detect mutations( I244T,33-34 insC, G170R, F152I, W108R,976delG and G190R) described as the most common.Clinical, biochemical, and radiological data were obtained by an information sheet review or provided by questionnaires sent to clinicians in charge of PHI patient. Results: Infantile onset (<1 years old) of symptoms occurred in 34 %, whereas 27% of patients experienced the first manifestation of disease at an adult age. Median age at diagnosis for infantile form was 7 years against 32 years for the adult form. The initial clinical presentation includes recurrent urolithiasis in 52% of cases, nephrocalcinosis in 38% and systemic oxalosis in 37%. ESRD was present at the time of diagnosis in 61 % of pediatric diagnoses patients versus 43 % of adult diagnosed patients. The diagnosis was based on urinary oxalate level in 24 h (32%), stone analysis (28%), family history (27%), urinary oxalate-to-creatinine molar ratio (25%), renal biopsy (14%) and urine glycolate excretion (3%).Molecular analysis revealed three mutations causing PH1 in Tunisia. The most predominant mutation was the Maghrebian”I244T” (30%). The 33_34insC was detected in 6% and the G190R was found in 3% of patients. About 60% of patients did not show any mutation of the seven studied, this would be to look for other mutation by sequencing. Conclusions: Our results underscore the heterogeneity of the HP 1, delayed diagnosis and poor prognosis. The disease is probably underdiagnosed, particularly in the adult form. The molecular identification could provide an accurate tool for prenatal diagnosis, preserve kidney function and prevent diagnosis of adult diagnosed patients in ESRD. MP062THE INTERLEUKIN-6 PATHWAY AND CARDIOVASCULAR DISEASE IN CKD: A MENDELIAN RANDOMIZATION STUDY Belinda Spoto Belinda Spoto 1 CNR-IFC, Reggio Calabria, Italy Daniela Leonardis Daniela Leonardis 1 CNR-IFC, Reggio Calabria, Italy Cristina Politi Cristina Politi 1 CNR-IFC, Reggio Calabria, Italy Anna Pisano Anna Pisano 1 CNR-IFC, Reggio Calabria, Italy Sebastiano Cutrupi Sebastiano Cutrupi 1 CNR-IFC, Reggio Calabria, Italy Alessandra Testa Alessandra Testa 1 CNR-IFC, Reggio Calabria, Italy Rosa Maria Parlongo Rosa Maria Parlongo 1 CNR-IFC, Reggio Calabria, Italy Graziella D'Arrigo Graziella D'Arrigo 1 CNR-IFC, Reggio Calabria, Italy Giovanni Tripepi Giovanni Tripepi 1 CNR-IFC, Reggio Calabria, Italy Francesca Mallamaci Francesca Mallamaci 1 CNR-IFC, Reggio Calabria, Italy Carmine Zoccali Carmine Zoccali 1 CNR-IFC, Reggio Calabria, Italy - On Behalf Of The MAURO Working Group - On Behalf Of The MAURO Working Group Abstract Introduction and Aims: High serum interleukin 6 (IL6) is a strong predictor of cardiovascular (CV) disease in large scale cohort studies in the general population. This cytokine is substantially increased in chronic kidney disease (CKD) patients but the relationship between IL6 and CV complications has been investigated just in one relatively small cohort study. Furthermore, whether the link between IL6 and CV disease in CKD is causal in nature is yet unknown. Methods: We studied the relationship between serum IL6 levels and background and incident CV events in a cohort of 755 CKD stage 2-5 patients (follow up: 31±10 months). We adopted the functional polymorphism (-174 C/G) in the promoter region of the IL-6 gene as un unbiased marker of long term exposure to circulating IL6 in order to investigate whether the IL6-CV events link is causal in nature. Results: High serum IL6 (> the median value=2.5 pg/mL) associated with background CV events both in unadjusted (OR: 2.14, 95% CI: 1.55-2.95, P<0.001) and multivariate adjusted analyses (OR: 1.58, 95%CI: 1.12-2.24, P=0.01). One hundred and twenty-four patients had incident CV events and the incidence rate of these events was by the 67% higher (HR=1.67 95% CI: 1.12-2.50; P=0.01) in patients with IL6 above the median as compared with patients with IL6 below this threshold in a fully adjusted analysis.Patients homozygous for the risk allele (G) of the -174 C/G polymorphism had higher levels of IL6 than those with other genotypes (GG: 2.90 pg/ml, IQR: 1.7-6.9 pg/ml; CG: 2.40 pg/ml, 1.6-3.8 pg/ml; CC: 2.50 pg/ml, 1.7-4.0 pg/ml, P=0.04). Homozygous GG patients had more frequently background CV events as well as a higher rate of incident events in unadjusted and fully adjusted (background CV events OR: 2.15, 95%CI: 1.15-4.0 and incident CV events HR:1.88, 95%CI: 1.02-3.44) analyses. Conclusions: High serum IL6 levels associate with background CV events and predict an increased risk of CV outcomes in stage 2-5 CKD. The parallel association of the risk allele for high IL6 in the -174 C/G polymorphism with background and incident CV events supports the contention that the serum IL6-CV events relationship is causal in nature. MP063PHENOTYPE-GENOTYPE CORRELATIONS IN PRIMARY HYPEROXALURIA TYPE 1: THE I244T, 33_34INSC AND G190R AGXT MUTATION Saoussen Mdimegh Saoussen Mdimegh 1 Biochemistry Department, Sahloul University Hospital, Sousse, Tunisia Ibtihel Ben Hadj Mbarek - Fredj Ibtihel Ben Hadj Mbarek - Fredj 1 Biochemistry Department, Sahloul University Hospital, Sousse, Tunisia Amira Moussa Amira Moussa 1 Biochemistry Department, Sahloul University Hospital, Sousse, Tunisia Asma Omezzine Asma Omezzine 1 Biochemistry Department, Sahloul University Hospital, Sousse, Tunisia Sameh Mabrouk Sameh Mabrouk 2 Pediatric Department, Sahloul University Hospital, Sousse, Tunisia Noura Zouari Noura Zouari 2 Pediatric Department, Sahloul University Hospital, Sousse, Tunisia Saida Hassayoun Saida Hassayoun 2 Pediatric Department, Sahloul University Hospital, Sousse, Tunisia Jalel Chemli Jalel Chemli 2 Pediatric Department, Sahloul University Hospital, Sousse, Tunisia Dorsaf Zellama Dorsaf Zellama 3 Nephrology Departement, Sahloul University Hospital, Sousse, Tunisia Abdelattif Achour Abdelattif Achour 3 Nephrology Departement, Sahloul University Hospital, Sousse, Tunisia Ali Bouslama Ali Bouslama 1 Biochemistry Department, Sahloul University Hospital, Sousse, Tunisia Saoussen Abroug Saoussen Abroug 2 Pediatric Department, Sahloul University Hospital, Sousse, Tunisia Abstract Introduction and Aims: Primary hyperoxaluria type I (PH1) caused by mutations in the AGXT gene, coding for the enzyme alanine-glyoxylate aminotransferase (AGT), resulting a wide spectrum of phenotypes ranging from renal failure in infancy to occur renal stones in late adulthood. We aimed to investigate a possible relationship between outcome of PH1 and AGXT mutation type. Methods: We present a retrospective study of 53 Patients who have been diagnosed with PH1, carriers of I244T, 33_34insC and G190R AGXT mutations. Ten patients, sibling of confirmed PH1 patients were included in the analysis. Male to female ratio was 0, 82(24/29).The clinical data were compiled and genetic testing was done by determining haplotype (Minor or Major) and mutations analysis using PCR/RFLP. Results: Haplotype analysis showed that each mutation co segregates with a specific haplotype. The Maghrebian mutation”I244T” was most frequently involved (80%), co segregates with the Minor allele, followed by the 33_34insC mutation (15%) and the G190R (6%) which cosegregates with the Major allele. Differences in age at diagnosis between PH1 mutation types was highly significant(p=0,012). Onset of symptoms occurred later in 33_34insC homozygotes (range: 0,2-61 years) compared with homozygote I244T (range: 0,1-40 years). The I244Thomozygotes were associated with slower progression to end-stage renal disease (ESRD) compared with 33_34insC homozygotes. Systemic manifestations were more frequently observed in the infantile form at the time of diagnosis and after evolution, in particular, ocular manifestations (11%), bone damaging (9%), cardiovascular (7%). In the I244T group, the mutation was found in 42 patients, 15 of them were diagnosed at an adult age and 11 after family screening. The median age of disease detection was 7 years. ESRD was reached in 45% homozygous patients. The I244T mutation has been associated with various renal symptoms: 47% urolithiasis, 31% nephrocalocinosis. In the 33_34insC group, a total of seven patients were homozygous, coding for a truncated protein, and 50% of them had infantile form, presented with nephrocalcinosis (60%) and five (62%) experienced ESRD. Three patients were carring the G190R mutation, all of them derived from the same family. It is presented with variable age at onset (range: 0,5- 16 years) and systemic manifestations at diagnosis and this especially for digestive (5%). No genotype phenotype correlation was seen. Most patients received chronic hemodialysis Conclusions: The HP1 is a rare disease which has the particularity to be more frequent in Tunisia. It is characterized by a great phenotypic variability therefore no consistent relationship between mutant genotype and phenotype. It is important to make an early diagnosis to prevent the devastating effects of systemic oxalosis. MP064MONOGENETIC AUTOINFLAMMATORY SYNDROMES AND KIDNEY FAILURE Katharina Hohenstein-Scheibenecker Katharina Hohenstein-Scheibenecker 1 Medical University of Vienna, Vienna, Austria Alice Schmidt Alice Schmidt 1 Medical University of Vienna, Vienna, Austria Abstract Introduction and Aims: For some years, the identification of genes involved in the modulation of inflammatory and apoptotic processes have allowed the delineation of a new group of diseases called "monogenetic autoinflammatory syndromes - MAISs".These syndromes are disorders of innate immunity characterized by recurring episodes of fever and systemic inflammatory symptoms, affecting the serosal surfaces, joints, skin and eyes and bear the serious risk of developing reactive systemic (AA) amyloidosis due to excessive production of serum amyloid-A (SAA). SAA is deposited in various organs, particularly the kidneys, with the consequent progressive development of proteinuria and kidney failure. Methods: We report three cases of kidney failure due to MAIS from our out-patient department. Results: Case Report 1A 54-year old dialysis patient with atrophic kidneys (reason unknown) underwent renal transplant in October 2012.A few days after discharge a fever of unknown origin occured with an increase of c-reactiv protein (CRP), creatinin, severe abdominal pain and diarrhoea without detection of pathogens. As familial Mediterranean fever (FMF) was suspected, patient underwent a colchicin therapy. The creatinin, CRP and SAA value decreased at normal range within 6 weeks of treatment. To confirm the diagnosed FMF, a mutation analysis was carried out. We found a heterocygous mutation in the MEFV gene, which confirms the FMF diagnosis.Case Report 2We report a now 26-year old patient, who was first admitted aged 5 months due to periodic fever. Additionally, patient’s history included polyarthritis, uveitis and a worsening of renal function over the years due to histological signs of sarcoidosis in the kidney. Different immunosuppressive treatments with steroids, azathioprin, methotrexat and cyclosporin did not prevent renal failure requiring dialysis in 2005. In 2010 our patient underwent renal transplantation. Despite standard triple immunosuppression, patient’s symptoms - especially uveitis - occurred even after transplantation whenever steroids were reduced. The initial diagnosis “juvenile sarcoidosis” was corrected to "Blau syndrom" due to mutation analysis carried out in April 2012 (mutation on NOD2 gene). Due to treatment with adalimumab since 2012, uveitis and polyarthritis are under control. Kidney function has remained stable with creatinin values within normal range up to this day.Case Report 3We report a woman born in 1971 who was diagnosed with reduced kidney function in 2008. In addition the patient has been suffering from cold-dependent urticaria and joint pain without known reason since childhood. The kidney biopsy shows signs of amyloidosis. Mutation analysis in June 2013 confirms the diagnose "Familial Cold-Autoinflammatory Syndrome - FACS". Due to treatment with anakinra, the cold-dependent urticaria and joint pain are eliminated, SAA is normalized and serum kreatinin is stabilized. Conclusions: Monogenetic Autoinflammatory Syndromes are rare causes of kidney failure. The detection by means of mutation analysis allowed a disease-specific therapy with stabilization of serum creatinin and SAA in addition to minimization of disease-specific symptoms even after renal transplantation. MP065HOMOZYGOUS SLC2A9 MUTATION CAUSING SEVERE HYPOURICEMIA AND REVERSIBLE EXERTIONAL ACUTE RENAL FAILURE Kostas G Stylianou Kostas G Stylianou 1 Heraklion University Hospital, Heraklion, Greece John Kyriazis John Kyriazis 2 General Hospital of Chios, Chios, Greece Ariadni Androvitsanea Ariadni Androvitsanea 1 Heraklion University Hospital, Heraklion, Greece Michael Tzanakakis Michael Tzanakakis 1 Heraklion University Hospital, Heraklion, Greece Eleftheria Maragkaki Eleftheria Maragkaki 1 Heraklion University Hospital, Heraklion, Greece John Petrakis John Petrakis 1 Heraklion University Hospital, Heraklion, Greece Stavros Stratakis Stavros Stratakis 1 Heraklion University Hospital, Heraklion, Greece Rafaela Poulidaki Rafaela Poulidaki 1 Heraklion University Hospital, Heraklion, Greece Eleftheria Vardaki Eleftheria Vardaki 1 Heraklion University Hospital, Heraklion, Greece Christina Petra Christina Petra 1 Heraklion University Hospital, Heraklion, Greece Spyros Statigis Spyros Statigis 1 Heraklion University Hospital, Heraklion, Greece Kostas Perakis Kostas Perakis 1 Heraklion University Hospital, Heraklion, Greece Eugene Daphnis Eugene Daphnis 1 Heraklion University Hospital, Heraklion, Greece Abstract Introduction and Aims: We present a case with homozygous mutation of glucose transporter SLC2A9 (GLUT9) causing severe hereditary renal hypouricemia (HRH)complicated by exercise induced acute renal failure (EIARF). Our findings provide further evidence for the key role played by GLUT9 in renal uric acid (UA) handling Methods: Identification of point mutations in coding exons in SLC2A9 gene. PCR and direct sequencing Results: A 30 year old man presented in the emergency department with severe back pain and nausea. The clinical examination revealed only mild dehydration with postural hypotension. His past medical history was remarkable only for repeated episodes of EIARF in the age of 15, 17, 28 and 30, which completely resolved after re-hydration. Family history was unremarkable. He was admitted in the renal ward for investigation and treatment of a new episode of EIARF. Blood tests were remarkable only for urea (52mg/dl), creatinine (2mg/dl) and uric acid (0.2mg/dl). Urine tests showed impaired concentrating ability with a specific gravity of 1009 despite dehydration. The fractional excretion of UA (FeUA) was remarkably high at 150%. Renal ultrasound revealed a mildly increased echogenicity. He was treated with normal saline which resulted in a rapid restoration of kidney function. A genetic analysis was performed to explore the possibility of renal hypouricemia as the triggering factor for EIARF. He was found to carry a homozygous deletion of exons 8-12 in the solute carrier family 2 member 9 (SLC2A9, OMIM 606142) gene coding for GLUT9 (Figure), along with 4 already known polymorphisms: c.49G>A (+/+), c.235T>C (+/+), c.288G>A (+/+), c.480T>C (+/+). This homozygous mutation was highly compatible with the phenotype of HRH. Patients usually present with symptoms such as nausea, abdominal pain, mild fever and acute renal failure after exercise. Two types of HRH have been described. Type-1, caused by SLC22A12 gene mutations coding for the urate transporter URAT1, is manifested by serum UA levels of 1-2mg/dl and FeUA of 40-90%. Type-2 is caused by SLC2A9 mutations and is characterized by very high FeUA (90-150%) and very low serum UA levels (<1mg/dl). Both types cause nephrolithiasis and EIARF, which has been attributed to ischemic kidney injury and aggregation of organic anions in the renal tubules. Conclusions: The reported mutation in SLC2A9 gene, causing severe loss of function of GLUT9, results in HRH associated with repeated episodes of EIARF Open in new tabDownload slide Open in new tabDownload slide MP066EFFICACY AND SAFETY OF ENZYME REPLACEMENT THERAPY (ERT) IN A COHORT OF KIDNEY TRANSPLANT RECIPIENTS WITH FABRY DISEASE Markus Cybulla Markus Cybulla 1 FGM, Center of Internal Medicine, Müllheim, Germany Michael West Michael West 2 Dalhousie University, Halifax, NS, Canada Kathy Nicholls Kathy Nicholls 3 Royal Melbourne Hospital & University of Melbourne, Melbourne, Australia Joan Torras Joan Torras 4 Hospital of Bellvitge IDIBELL, Barcelona, Spain Pablo Neumann Pablo Neumann 5 Servicio de Nefrologia, Hospital Italiano de La Plata, La Plata, Argentina Gere Sunder-Plassmann Gere Sunder-Plassmann 6 Medical University of Vienna, Department of Nephrology and Dialysis, Vienna, Austria Sandro Feriozzi Sandro Feriozzi 7 Belcolle Hospital, Viterbo, Italy On Behalf Of The Fabry Outcome Survey Renal Working Group On Behalf Of The Fabry Outcome Survey Renal Working Group Abstract Introduction and Aims: Enzyme replacement treatment (ERT) was introduced in 2001 as a treatment option for patients with Fabry disease. The Fabry Outcome Survey (FOS, sponsored by Shire) is a worldwide outcome database, which was established to monitor the long-term effectiveness and safety of ERT with agalsidase alfa. This study presents clinical data of a follow-up analysis in kidney transplant recipients (KTRs) with Fabry disease. Methods: The effects of long-term ERT with agalsidase alfa (0.2mg/kg every other week; Shire) on renal outcomes were analyzed using Fabry patients’ data in FOS (data extraction November, 2012). Estimated glomerular filtration rate (eGFR) was calculated using the Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) equation. For the purposes of this analysis, blood pressure values above normotensive levels (≥120/80 mmHg) were considered hypertensive. Results: Renal function was analysed in 51 KTRs (47 treated with ERT before or after transplant surgery; 4 untreated). For the 47 treated KTRs, the median (range) time on ERT until the last visit was 2.9 (0.0-13.4) years. Open in new tabDownload slide Open in new tabDownload slide During the observation time (start of ERT and last visit), the average annualized eGFR slopes were not significantly different for treated and untreated patients, although there was a numerically steeper decline in untreated than in treated male patients, whose renal function was relatively stable. The impact of proteinuria on outcomes was difficult to assess as patients’ preoperative and donors’ proteinuria histories were not known. Finally, dialysis treatment before transplantation had no negative impact on later graft function. Only minor side effects or infusion-related reactions were reported. Conclusions: Conclusions from this study should be made carefully because of the low number of patients, particularly in the KTR without ERT group. Our findings, however, suggest that graft function in KTR Fabry patients remains stable under ERT andthat the treatment seems to be well tolerated. MP067POSSIBLE INVOLVEMENT OF MICRORNAs IN VASCULAR DAMAGE IN EXPERIMENTAL CHRONIC KIDNEY DISEASE Valerie Metzinger-Le Meuth Valerie Metzinger-Le Meuth 1 Université Picardie Jules Verne, Amiens, France Fatiha Taïbi Fatiha Taïbi 2 UPJV, Amiens, France Eleonore M'Baya-Moutoula Eleonore M'Baya-Moutoula 2 UPJV, Amiens, France Loic Louvet Loic Louvet 2 UPJV, Amiens, France Ziad Massy Ziad Massy 2 UPJV, Amiens, France Laurent Metzinger Laurent Metzinger 2 UPJV, Amiens, France Abstract Introduction and Aims: Chronic kidney disease (CKD) is associated with vascular calcificationsand atherosclerosis. There is a need for novel predictors to allow earlier diagnosis of these disorders, predict disease progression, and improve assessment of treatment response.We focused on microRNAs since they are implicated in a variety of cellular functions in cardiovascularpathology. Methods: We examined changes of microRNA expression by RT-qPCR in aortas of CKD and non-CKD wild type mice and apolipoprotein E knock-out mice, respectively. Results: Both vascular smooth muscle-specific miR-143 and miR-145 expressions were decreased in states of atherosclerosis and/or CKD or both, and the expression level of protein target Myocardin was increased. The inflammatory miR-223 was increased in more advanced stages of CKD, and specific protein targets NFI-A and GLUT-4 were dramatically decreased. Expression of miR-126 was markedly increased and expression of protein targets VCAM-1 and SDF-1 was altered during the course of CKD.The drug sevelamer, commonly used in CKD, corrected partially these changes in microRNA expression, suggesting a directlink between the observed microRNA alterations and uremic vascular toxicity.Finally, miR-126, -143 and -223 expression levels were deregulated in murine serum during the course of experimental CKD. Conclusions: In conclusion, these miRNAs could have role(s) in CKD vascular remodeling and may therefore represent useful targets to prevent or treat complications of CKD. MP068ASSOCIATION OF FOCAL AND SEGMENTAL GLOMERULOSCLEROSIS AND CHARCOT-MARIE-TOOTH DISEASE: IDENTIFICATION OF A NEW MUTATION IN INVERTED FORMIN 2-GENE Laila-Yasmin Mani Laila-Yasmin Mani 1 University Clinic for Nephrology, Hypertension and Clinical Pharmacology, Bern, Switzerland Daniel Sidler Daniel Sidler 1 University Clinic for Nephrology, Hypertension and Clinical Pharmacology, Bern, Switzerland Bruno Vogt Bruno Vogt 1 University Clinic for Nephrology, Hypertension and Clinical Pharmacology, Bern, Switzerland Abstract Introduction and Aims: We report the case of an adult patient having received a kidney transplant during childhood for focal and segmental glomerulosclerosis (FSGS) diagnosed with Charcot-Marie-Tooth (CMT) disease. A heterozygous mutation of the inverted formin 2 (INF2)-gene was identified that has not been described yet, occurring in the gene region where mutations with the identical clinical context were found recently (Boyer et al. 2011). Methods: A 43-year old female patient is regularly followed since kidney transplantation at the age of 17 years for end stage renal disease (ESRD) due to FSGS with an uncomplicated clinical evolution. Nephrotic-range proteinuria was first detected during medical school screening at age 15, associated with impaired renal function and arterial hypertension. Renal biopsy at the age of 16 years was consistent with a FSGS lesion. Kidney disease progressed rapidly under short treatment with ciclosporin and steroids and reached a terminal level at age 16 with initiation of chronic hemodialysis. In addition, the patient suffered from hereditary motor and sensory neuropathy type I (CMT disease) diagnosed at the age of nine years; electromyoneurogram showed slowed nerve conduction velocities of N. peroneus; a sural nerve biopsy at the age of nine years revealed marked reduction in myelinized nerve fibers and the presence of mast cells; genetic testing was not performed. The phenotype included stumbling and falls since the age of three years and progressive distal muscle weakness of upper and lower extremities leading to foot (pes equinovarus and cavus) and hand deformities requiring repeated orthopedic interventions. Symmetric sensorineural hearing loss at 2000 Hz was objectified at age 19 leading to bilateral treated deafness since the age of 38 years. Familial history revealed no cases in family members; there are no siblings or offspring of the index case. Review of the native kidney biopsy with performance of electron microscopy confirmed the FSGS lesion, not otherwise specified (NOS)-variant. In the clinical context of FSGS with CMT disease, a mutation in the INF2 gene was suspected. Results: Direct sequencing of exons 2 and 3 of the INF2 gene revealed the heterozygous mutation c.367_375del (p.Gly123_Val125) with deletion of nine base pairs and three amino-acids in exon 2 respectively. This mutation has not been previously described and localizes in the gene region where mutations in cases with the association of CMT disease and FSGS were described recently (N-terminal diaphanous-inhibitory domain, second and third armadillo repeats; exon 2, nucleotides 300-500). Conclusions: We identified a new heterozygous likely spontaneous mutation in the INF2 gene. As localized in the gene region where mutations leading to this phenotype have been described until now, it is most likely to explain the renal and neurological presentation in this patient. It can however not be excluded that the mutation found is merely responsible for the renal disease as genetic testing for classical genes for CMT disease has not been performed. MP069CKD IN TSC PATIENTS WITH DIFFERENT RENAL PHENOTYPES Natalya Nikolskaya Natalya Nikolskaya 1 Sussex Kidney Unit, Brighton, United Kingdom Jane A Cox Jane A Cox 1 Sussex Kidney Unit, Brighton, United Kingdom John C Kingswood John C Kingswood 1 Sussex Kidney Unit, Brighton, United Kingdom Abstract Introduction and Aims: Renal disease due to haemorrhage and cancer is a major cause of morbidity and mortality in adult patients with tuberous sclerosis complex (TSC). However, chronic kidney disease (CKD) has not been systematically studied in these patients.The role of different renal phenotypes, as is the influence of gender factors, hereditary factors, surgical treatment and the combinations of these factors in progression of renal failure is still unclear. To determine the influence of renal phenotypes in patient with TSC and analyze the risk factors for development and progression of CKD. Methods: A database of 251 patients with TSC was analyzed. 113 adult patients with different renal phenotypes had at least one serum creatinine measurement. Estimated GFR were calculated using the 4-variable MDRD equation. Results: The sample consisted of three groups: 98 patients with angiomyolipomata (AML), average eGFR was 76.8 ml/min/1.73 m², mean age 38.5 years; 6 with polycystic kidney disease (PKD), average eGFR was 27 ml/min, mean age 32.3 years; and 9 with both AML and PKD average eGFR was 62.8 ml/min/1.73 m², mean age 42.8 years.Renal phenotype, family history, previous surgical renal intervention and female sex were all associated with a significant difference in GFR. Conclusions: Monitoring renal function is vital in TSC patients, as pre-emptive treatment of problems may preserve GFR. MP070URINARY microRNA-21 EXPRESSION IN NEPHROPATHIES Alexey Smirnov Alexey Smirnov 1 First Pavlov State Medical University of St. Petersburg, Saint Petersburg, Russian Federation Mikhail Zarayski Mikhail Zarayski 1 First Pavlov State Medical University of St. Petersburg, Saint Petersburg, Russian Federation Ivan Kayukov Ivan Kayukov 1 First Pavlov State Medical University of St. Petersburg, Saint Petersburg, Russian Federation Hanna Karunnaya Hanna Karunnaya 1 First Pavlov State Medical University of St. Petersburg, Saint Petersburg, Russian Federation Vassiliy Sipovski Vassiliy Sipovski 1 First Pavlov State Medical University of St. Petersburg, Saint Petersburg, Russian Federation Larisa Kukoleva Larisa Kukoleva 1 First Pavlov State Medical University of St. Petersburg, Saint Petersburg, Russian Federation Vladimir Dobronravov Vladimir Dobronravov 1 First Pavlov State Medical University of St. Petersburg, Saint Petersburg, Russian Federation Abstract Introduction and Aims: MicroRNAs (miRNAs) are a class of noncoding RNA acting at a post-transcriptional level to control the expression of large sets of target mRNAs. While there is evidence that miRNAs deregulation plays a causative role in various complex disorders, their role in fibrotic kidney diseases is largely unexplored. However some date suggest, that increased circulating miR-21 levels are associated with kidney fibrosis. In this regard, we attempted to compare the level of miR-21 expression in the urine with signs of kidney lesions in patients (Pts) with nephropathy (Neph). Methods: Seventeen Pts with different Neph confirmed by kidney biopsy were examined: IgA nephropathy (n=4), focal segmental glomerulosclerosis (n=3), minimal change disease (n=2), membranoproliferative glomerulonephritis (n=2), AL amyloidosis (n=2), other (n=4). miR-21 expression in the urine was determined by a RT-PCR assay (EvaGreen florescence dye) and calculated using the 2-delta Ct protocol. In Pts glomerular filtration rate (GFR) was assessed by creatinine clearance (CCr), Cockcroft&Gault, MDRD7 and CRD-EPI formulas. Daily urinary protein (UPE, g/24), creatinine (UCr, mmol/24), urea (UUr, mmol/24), potassium (UK, mmol/24), sodium (UNa, mmol/24), calcium (UCa, mmol/24), chloride (UCl, mmol/24) and inorganic phosphorus (UP, mmol/24) has been establish. Fractional excretion (relative to CCr; FE, %) of all the previously mentioned chemical components of urine, except for protein and creatinine, was calculated. Total protein (TP, g/l) and albumin (Alb, g/l) serum concentrations also been taken into account. Percent of total sclerosed glomeruli and glomeruli with segmental sclerosis were calculated in the kidney bioptates (late microscopy). The degree of tubular atrophy (ТА), diffuse (DF) and segmental (SF) interstitial fibrosis were evaluated semi-quantitatively on a scale from one to three stages (1 - mild changes, 2 - moderate changes, 3 - severe changes). miR-21 expression in the urine of healthy donors (n=11) was taken as control (C). All date are presented as mean±SE. Unpaired Student t-test and Spearman rank correlation coefficient (RS) were used. Results: The urine levels of miR-21 expression in Pts with Neph were significantly higher than in C (0.370±0.0002 vs 0.031±0.017; respectively, p=0.0011). There was strong positive correlation between urine miR-21 (RS=0.570, p=0.017) in Pts. In Neph the value of RS (-0.463, p=0.061) between miR-21 and Alb did not achieve statistical significance. In third stage (0.568±0.122; n=7) of TA urine miR-21 was significant more than second (0.202±0.067; n=7, p=0.021). Pts with first stage of TA were only three, that did not allow perform adequate statistical comparisons. There were no any significant associations between urine miR-21 and other functional or morphological parameters studied. Conclusions: These data suggest that the level of miR-21 expression in the urine to a certain degree can be associated with the severity of renal damage in patients with nephropathies. However, to clarify the role of miR-21 in the development and progression of renal lesions further investigations are needed. © The Author 2014. Published by Oxford University Press on behalf of ERA-EDTA. All rights reserved
TI - GENETIC DISEASES AND MOLECULAR GENETICS
JO - Nephrology Dialysis Transplantation
DO - 10.1093/ndt/gfu162
DA - 2014-05-01
UR - https://www.deepdyve.com/lp/oxford-university-press/genetic-diseases-and-molecular-genetics-8JTTAcA6OB
SP - iii339
EP - iii350
VL - 29
IS - suppl_3
DP - DeepDyve
ER -