bluuebunny/crossref_metadata_2025_split huggingface dataset:
#2928 Fabry disease may imbalance integrin α3 related to endothelial dysfunction
Authors: Santana Aline Aparecida , Gregorio Paulo Cézar , Da Cunha Regiane Stafim , de Almeida Guilherme Miniskiskosky , Azevedo Carolina Amaral Bueno , Franco Célia Regina Cavichiolo , Barreto Fellype , Stinghen Andréa E M 
Published: May 2024

Abstract

Background and Aims
Fabry Disease (FD) is a rare disease characterized by a mutation in the GLA gene and consequently by the functional anomaly of the α-Galactosidase A (α-Gal) enzyme. The lack of this functional enzyme leads to lysosomal accumulation of glycosphingolipids, mainly globotriaosylceramide (Gb3), in different cell types, causing several symptoms throughout the body, which aggravate during the patient's longevity. We already demonstrated that exposure of endothelial cells to chloroquine, to mimic FD effects, leads to lysosomal accumulation in human endothelial cells. In the present study, we analyzed the endothelial changes in ultrastructural morphology and cell adhesion under evaluation of Integrin α3 expression, an important integrin that stabilizes cell adhesion and is involved in several signaling pathways in endothelial cells.

Method
Human endothelial cells (EA.hy926, ATCC CRL-2922) were exposed to chloroquine diphosphate (0.5 µg/mL) for 72 hours. The ultrastructural morphology was analyzed through scanning electron microscopy (SEM) and the Integrin α3 expression was analyzed through Western Blot and RT-qPCR.

Results
Through SEM (Fig. 1C-F) it was observed that after the exposure to chloroquine diphosphate, there was a decrease in cell expansion and adhesion and an increase in vesicular content. Curiously, the analysis of the Integrin α3 gene (ITGA3) and protein expression (Fig. 1A-B) demonstrated a significant increase (P &lt; 0.05) in cells that were exposed to chloroquine diphosphate compared to non-exposed cells (control cells).

Conclusion
The in vitro condition of the FD in human endothelial cells demonstrated an increase in Integrin α3; however, there was a decrease in cell adhesion and expansion. The evaluation of the Integrin α3 expression could be important for monitoring the endothelial dysfunction in FD, as it is intrinsically involved in cell adhesion, remodeling, and fibrosis. These important changes can lead to renal impairment, proteinuria, and decreased glomerular filtration rate.
#2416 Bicc1 and PKD: linking RNA metabolism to kidney disease
Authors: Constam Daniel B 
Published: May 2024

Abstract

Background and Aims
Mutations in PKD1 or PKD2 lead to autosomal dominant polycystic kidney disease (PKD) by allowing urine flow-sensing primary cilia to activate tissue growth signals. However, the responsible cilia-associated factors and a strategy to block them and cure PKD are elusive. Cilia also specify organ laterality. In this process, flow-stimulated cilia activate the cytosolic protein Bicaudal-C (Bicc1) to curb the translation and/or the stability of specific target mRNAs. To identify candidate regulatory factors, we investigated if Bicc1 ribonucleoprotein particle (RNP) formation involves known proteins linked to cilia and cystic kidney diseases.

Method
To address this question, we here used AlphaFold structure modeling of protein complexes, combined with in vitro reconstitution assays and live imaging of Bicc1 RNP assembly. To begin to validate these interactions, we tested by genetic approaches if specific interacting factors also regulate Bicc1 targets in cells and mice.

Results
We found that multivalent interactions with structured domains of the ankyrin repeat and SAM domain containing (ANKS) 3 and their allosteric regulation by the related ANKS6 protein inhibit or restore Bicc1 access to target mRNAs, respectively. Concomitantly, ANKS3 alone increased the solubility and fluidity of Bicc1, whereas co-recruitment of ANKS6 had the opposite effect, liberating Bicc1 to self-polymerize and to induce phase-transitioning of target mRNAs in gel-like meshworks.

Conclusion
Perturbations in Bicc1-ANKS3-ANKS6 RNP dynamics and altered translation of Bicc1 target mRNAs emerge as likely culprits regulating cyst formation downstream of ciliary defects. ANKS6 can be sequestered in cilia by inversin, and mutations in these proteins provoke kidney cysts in nephronophthisis patients. Therefore, besides systematically profiling Bicc1 RNPs, future studies should test if ANKS6 mediates their dynamic regulation by flow-stimulated cilia.
#3020 A novel copy number analysis identifies human patients with NPHP1 whole gene deletions in previously genetically unsolved cases
Authors: Sayer John , Olinger Eric , Wilson Ian 
Published: May 2024

Abstract

Background and Aims
Genome sequencing is rapidly becoming a routine approach to investigating inherited human diseases in clinical practice. Nevertheless, data generated from genome sequencing requires extensive bioinformatic analysis and filtering of variants to determine pathogenicity and remains challenging. Despite well-established workflows and software in place to process raw data, the analysis of genome sequencing data is still complex. Copy Number Variations (CNVs) are increasingly recognised to be pathogenic, especially where this leads to a whole gene being deleted. Software tools to decipher CNVs across genome sequencing datasets are being utilised but are not completely accurate.

Method
Using a bespoke CNV analysis of Genomics England 100,000 Genomes Project (100 kGP) WGS data from 11754 parent-child trios, we searched for homozygous deletions overlapping with annotated genes, across the 22 autosomes.

Results
Homozygous ∼150 kb whole gene deletion of NPHP1-MALL on chromosome 1, which aligned with a known region containing segmental duplications (SD), was the most frequent observation among all autosomal recessive (AR) OMIM-morbid genes. Clinical phenotypes were consistent with this genotype including nephronophthisis and retinal dystrophy. We extended our analysis to look for NPHP1 whole gene deletions in 18877 additional singletons within the 100 kGP. This reverse genetics approach identified additional patients, up to this point genetically unsolved, with whole gene deletions and matching phenotypes. The frequency of heterozygous NPHP1 whole gene deletion was assessed in a control cohort of 15440 cancer patients within 100 kGP and was found to be 0.35%.

Conclusion
In conclusion, NPHP1 was the most frequently observed AR OMIM-morbid gene knockout in our analysis. Homozygous whole gene deletions in NPHP1 may be easily missed using standard WGS pipelines which don't assess fully for CNVs and a targeted read-depth approach to identify CNVs in parent-patient trios and singletons is valuable in identifying these deletions.
#1699 The role of interleukin 12 cytokine family in autosomal dominant polycystic kidney disease physiopathology
Authors: Ene Corina-Daniela , Penescu Mircea , Nicolae Ilinca 
Published: May 2024

Abstract

Background and Aims
Autosomal dominant polycystic kidney disease (ADPKD) is an inherited, complex disease, characterized by cysts formation and phenotypic changes. ADPKD physiopathology is under research, the role of proinflammatory chemokines responsible for perpetuating the cycle of chronic inflammation leading to kidney damage being intensively studied. We aimed to explore the individualized contribution of IL-12 interleukin family members in ADPKD physiopathology.

Method
We developed a prospective, case control study, in Carol Davila Clinical Hospital of Nephrology, including a group of 66 ADPKD subjects (56.1 years old, men:women 35:32), and a group of 40 healthy subjects, similar as sex, mean age. ADPKD diagnosis was based on familial history, clinical exam and CT or MRI scan. We excluded from the study subjects that presented cysts in other organs except the kidney, with eGFR&lt;60 mL/min/1.73 mp, with history of hematuria, cysts infection, urinary tract infection, renal lithiasis, with unstable blood pressure in the last 6 months.
Laboratory tests: serum levels of monomers IL-12 p40 and IL-20 p35 and heterodimers IL-12 p70, IL-23, IL 35, assessed by ELISA method.
Statistical analysis. All the results were analyzed using IBM SPSS Statistics 2015 and were presented as average value ± standard deviation. p &lt; 0.05 was considered with statistical significance.

Results
In ADPKD patients, we detected altered production of IL-12 cytokine, with low production of IL-12 p35 and IL-35, respectively high production of IL-12 p70, IL-23 and IL-12 p40 versus control. IL-12 p35 and IL-35 had statistically significant lower levels in ADPKD patients with eGFR&lt;90 ml/min/1.73 mp, when compared with ADPKD patients with eGFR&gt;90 ml/min/1.73 mp (Table 1). IL-12 p70, IL-23 and IL-12 p40 had statistically significant higher levels in ADPKD patients with eGFR&lt;90 ml/min/1.73 mp, when compared with ADPKD patients with eGFR&gt;90 ml/min/1.73 mp (Table 1). IL-12p70, IL-23 and IL-12 p40 had a negative significant correlation with eGFR, while IL-12p35 and IL-35 had a positive significant correlation with eGFR. Though, IL-12p35 and IL-35 serum levels correlated negatively with IL-12p70, IL-23 and IL-12 p40 serum levels in ADPKD patients.

Conclusion
The results of the present study suggest that ADPKD is a cellular source of IL-12 family, cytokines produced by antigenic stimulation. IL-12 family programming is abnormal in ADPKD patients. We detected synergic and divergent action of IL-12 cytokine family members, relevant for ADPKD physiopathology.
#1493 Upregulated miR-205 as a biomarker of diet-induced obesity-related glomerulopathy
Authors: Navarro Maru , López-Martínez Marina , Armengol Maria Pilar , Pey Irina , Farré Xavi , Rodríguez-Rodríguez Paula , Ferrer Mireia , Porrini Esteban , Luis-Lima Sergio , Martin Laura Díaz , Ampudia Rosa 
Published: May 2024

Abstract

Background and Aims
Obesity is a modifiable risk factor for the development and progression of chronic kidney disease (CKD). Pathways through which obesity might cause renal disease are not completely understood, as not all obese subjects develop CKD. In terms of deepening the knowledge of the preliminary steps of obesity-related renal damage, our group has developed a diet-induced obese murine model with obesity related glomerulopathy (ORG). The objective of this study was to investigate ORG at a cellular, structural and transcriptomic level as a way to discover the triggering pathogenic phenomena of ORG and evaluate potential biomarkers.

Method
Thirty Wistar rats were randomized into two groups: control rats (n = 15), which were fed with standard diet (SD) and study rats (n = 15), which were fed with high-fat diet (HFD). After 10 weeks, weight, parameters of kidney function, renal histological features, transcriptomic changes, miRNA and mRNA isolation were compared.

Results
HFD gained significantly more weight (55.8%) than SD(29.2%), p = 0.001. Albuminuria was also significantly higher in HFD (10,384.04) compared with SD (5,845.45), p = 0.001. HFD rats showed typical lesions of early-stages of ORG, with a predominance of mesangial matrix increase (MMI) and podocyte hypertrophy (PH). All histologic lesions correlated with genes differentially expressed (DE) in kidneys of HFD group and, both MMI and PH, also correlated with specific miRNAs DE in the urine of HFD group. The functional analysis showed 4 miRNAs DE in the kidneys of HFD group that negatively regulates PTEN gene, which promotes podocyte endocytosis of lipids in ORG. The electronic microscope confirmed the spaces of lipid vacuoles in the podocytes of HFD rats. Between those 4 miRNAs DE, miR-205 was also found to be upregulated in the urine of HFD group.

Conclusion
Wistar rats fed with HFD for 10 weeks developed early-stages of ORG, with a specific targetome of miRNAs and gene expression. The upregulation of miR-205 in kidney and its isolation in urine is associated with lipid endocytosis of podocytes, which takes place in the early-stages of ORG and could become a plausible biomarker of early-stages of ORG and open new avenues for future therapeutics research.
#2858 Immunohistochemical study reveals increased GLUT1 expression in proximal tubules of patients with acute interstitial nephritis
Authors: Shabaka Amir , Garcia-Fernandez Eugenia , Yébenes Laura , Alonso Patricia , Mendoza Ana , Triska Eugenia Landaluce , León-Póo Mariana , Garcia-Bermejo Laura , Juarez Gema Maria Fernandez 
Published: May 2024

Abstract

Background and Aims
Interstitial inflammation in acute interstitial nephritis can advance into tubular epithelial cells and produce proximal tubular dysfunction, which may be clinically expressed as renal glycosuria. Previous studies have shown that the presence of renal glycosuria in acute kidney injury is a diagnostic clue for acute interstitial nephritis (AIN) with an excellent specificity. Sodium-independent glucose transporter 1 (GLUT1) is localized in the basolateral membrane of epithelial cells along the entire length of the proximal tubule, mainly in the S3 segment, and it is involved in the transcellular glucose transport. The aim of this study was to examine renal expression of GLUT1 in the proximal tubules of patients with biopsy-proven AIN.

Method
We identified patients with biopsy-proven AIN. After excluding patients with diabetes mellitus and patients who had received steroid treatment previous to the kidney biopsy, we selected 10 patients with biopsy-proven AIN, and we selected 10 patients with acute tubular necrosis (ATN) matched for age, proteinuria and glomerular filtration rate as the control group. An immunohistochemical study for GLUT1 was performed on the biopsies of patients with AIN and ATN. Staining intensity was graded as 0 (negative), 1+ (weak), 2+ (moderate), and 3+ (strong). GLUT1 expression was defined as focal when &lt;50% of epithelial cells stained positive, and diffuse when &gt;50% of the tissue showed strong staining.

Results
Renal glycosuria was present in 60% of the selected patients with AIN, with a median glycosuria of 150 mg/dl (IQR 0-162 mg/dl), while only 1 patient (10%) with ATN presented glycosuria (p = 0.037). All patients with AIN showed moderate or strong cytoplasmic GLUT1 staining in the epithelial cells of proximal tubules, which was significantly more intense (2+; 20% vs 11%, 3+; 80% vs 33.3%, p = 0.008) and diffuse (80% vs 22.2%, p = 0.002) than in patients with ATN. There was a tendency for positive correlation between the concentration of glycosuria and the extension of GLUT1 staining (r = 0.316, p = 0.078).

Conclusion
We found higher expression of GLUT1 in the cytoplasm of proximal tubular cells in patients with AIN in comparison to ATN. These findings suggest that proximal tubular injury plays a key role in the pathogenesis of AIN, leading to renal glycosuria in some cases. We hypothesize that the localization of strong GLUT1 staining in the cytoplasm of proximal tubular cells may indicate a misallocation of GLUT1 from plasma membrane where it is functional and a reversed cell polarity which might account for the observed renal glycosuria.
#1978 Proteomic analysis of exosomes derived from mesangial cells treated with galactose-deficient IgA1
Authors: Narasimhan Gayathri , Ebefors Kerstin , Boi Roberto , Johansson Alva , Nyström Jenny 
Published: May 2024

Abstract

Background and Aims
IgA nephropathy (IgAN) is an autoimmune disease characterized by IgA deposits that build up in the kidneys, inflame and damage glomeruli causing hematuria and proteinuria. It is a leading cause of chronic kidney disease, which often results in end-stage kidney disease. There is no disease specific treatment for IgAN. The glomerulus, in which IgA1 lacking galactose residues (gdIgA1) are deposited, consists of mesangial cells, podocytes, and endothelial cells. Crosstalk between the cell types allows the glomerulus to act as a unit and react to its dynamic environment. There is very little evidence on mesangial cell derived exosomes and their cargo to understand the pathological crosstalk signaling in the glomerulus of IgAN. We isolated exosomes from mesangial cells that were treated with patient-derived gdIgA1 and IgA1 complexes. Proteomic analysis was performed to identify the exosome cargo and delineate the signaling pathways leading to maladaptive paracrine crosstalk. From our study, we aim to understand the molecular targets for therapeutic strategies in IgAN.

Method
Human mesangial cells were treated with/without IgA1, gdIgA1 or IgG for 24 h. Cell culture supernatant was collected for exosome isolations. Exosomes were analyzed by size, morphology and number using TEM and NanoFCM. Exosome protein extracts were processed for LC-Tandem mass spectrometry analysis using filter-aided sample preparation protocol. Proteomic analysis was performed using Qlucore omics explorer and pathway analysis using Ingenuity Pathway Analysis (IPA). Exosome marker expressions such as CD63, CD81 and FLOT1 and proteome validation was performed using Western blotting. Statistical analysis was performed using GraphPad Prism and values &lt;0.05 were considered significant.

Results
We confirmed the release of exosomes by mesangial cells in response to IgA1/gdIgA1 complexes. Exosomes were fewer in unstimulated cells as compared to IgA1/gdIgA1 treated cells. They showed round morphology with a dense core and uniform size distribution. Exosomes from treated cells showed a mean size of about 75 nm and concentration in the range of 1-2 × 10^10 particles/ml. While untreated cells showed a mean size of about 100 nm and concentration of about 1 × 10^9 particles/ml. We confirmed the protein expression of exosome markers such as CD63, CD81 and FLOT1.
For the first time, we demonstrated global proteomics on exosomes from mesangial cells followed by pathway analysis. We have identified 89 unique proteins and pathways in exosomes derived from gdIgA1 treated mesangial cells that could mediate pathological signaling in the glomerulus in comparison to exosomes derived from IgA1 treated mesangial cells.

Conclusion
Our study provides an understanding of the pathological crosstalk signaling through exosomes and key molecular targets for IgAN.
#2549 Real world data of dapagliflozin effect on proteinuria in primary glomerulopathies in Croatia
Authors: Brinar Ivana Vukovic , Laganovic Mario , Zagorec Nikola , Šimunov Bojana , Radić Josipa , Gulin Tonko , Altabas Karmela , Bubic Ivan , Šimundić Tihana , Ivandic Ema , Pranjić Ita Jelić , Horvatic Ivica 
Published: May 2024

Abstract

Background and Aims
SGLT2 inhibitors, namely dapagliflozin, proved to slow progression of kidney disease in randomized clinical trials. Our aim was to assess real world efficacy of dapagliflozin on proteinuria level in primary glomerulopathies (GN).

Method
adult patients with biopsy proven glomerulonephritis from all university hospitals in Croatia who started dapagliflozin as a standard of care were enrolled. Data were analyzed if patients had at least six months of follow up. None of the patients were on immunosuppression. Data are shown as N (%) or median with interquartile range (IQR) and accompanied p levels.

Results
Overall 118 patients, predominantly male 84 (71.8%) of average age of 52 years (min–max 20–76), were enrolled. Most of the patients had IgA nephropathy (IgAN) (N = 55;46.6%), followed by membranous nephropathy (N = 27;22.9%), FSGS (N = 19;16.1%), membranoproliferative GN (N = 8;6.8%), Alport syndrome (N = 8;6.8%). 95.8% of patients were on ACE inhibitors or angiotensin receptor blockers and 10.7% were on mineralocorticoid receptor inhibitors and RAAS blockage was not significantly upscaled during follow up. During the median follow up of 12.2 (7.5-18.6) months, proteinuria was significantly reduced from 1139 (534–2265) to 701 (305-1543) mg/dU; p &lt; 0.001. When divided by glomerulonephritis type, statistically significant reduction of proteinuria was observed in patients with IgAN and Alport syndrome, while trend of reduction of proteinuria was observed in all types of GN. There was a slight reduction in eGFR levels (54 (43-78) vs. 52 (39–73) ml/min/1,73 m²; p &lt; 0.001) and rise in creatinine values (122 (89–149) vs. 128 (91–159) μmol/l; p &lt; 0.001). There was a significant reduction in systolic (130 (120-140) vs.125 (115-134) mmHg; p &lt; 0.001) and diastolic blood pressure (80 (75-90) vs. 80 (70-85) mmHg; p &lt; 0.001). Reduction of uric acid level was noticed (377 (313–460) vs.343 (294–422); p &lt; 0.001), while there was no effect on LDL levels (2.6 (2.12–3.53) vs.2.3 (1.9–3.30); p = 0.073). Dapagliflozin was well tolerated, one patient discontinued the therapy due to intolerance, one due to urinary infection and two due to relapse of GN which needed immunosuppressive therapy. One patient was excluded from analysis due to non-adherence.

Conclusion
SGLT2 inhibitor dapagliflozin has significant effect on reduction of proteinuria in patients with primary glomerulonephritis. Our data support a growing body of evidence that SGLT2 inhibitors should be used as a standard of care in patients with glomerular kidney diseases, especially IgAN. Future research should address earlier use of SGLT2 inhibitors during the treatment of glomerulonephritis alongside immunosuppressive therapy as well as look into the effect of SGLT2 inhibitors on mesangial cells.