Understanding the beginning of diabetic nephropathy with electric potentials and repulsion forces around the renal glomerulus in the human kidney

2017 ◽  
Author(s):  
Huber Nieto-Chaupis
Keyword(s):  
Diabetic Nephropathy ◽  
Human Kidney ◽  
Renal Glomerulus ◽  
Electric Potentials

scholarly journals SO026HUMAN KIDNEY PROXIMAL TUBULAR 3D SPHEROIDS TO STUDY THE ROLE OF ADAM17 IN DIABETIC NEPHROPATHY

2020 ◽  
Vol 35 (Supplement_3) ◽  
Author(s):  
Vanesa Palau ◽  
Bramasta Nugraha ◽  
Maximilian Emmert ◽  
Simon Hoerstrup ◽  
Julio Pascual Santos ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cell Culture ◽  
Diabetic Nephropathy ◽  
High Glucose ◽  
Human Kidney ◽  
Collagen Iv ◽  
Control Group ◽  
Wild Type ◽  
Factor Α ◽  
3D Spheroids

Abstract Background and Aims ADAM17 is a disintegrin and metalloproteinase initially described to cleave the tumor necrosis factor α (TNFα). Currently, it is known that it can also release ectodomains of a diverse variety of molecules such as, transforming growth factor α (TGFα), L-selectin, and angiotensin-converting enzyme 2 (ACE2). It has been shown that ADAM17 protein expression increases in kidney mesangial cells after incubation with high glucose media mimicking what has been observed in diabetic patients and experimental models of diabetic nephropathy. We now studied the effect ADAM17 deletion on human kidney cells (HKC-8) in a 3D spheroids in vitro cell culture incubated with high glucose, low glucose and mannitol medium resembling the in vivo human kidney diabetic environment. Method ADAM17 deletion was performed using the CRISPR/Cas9 technology. HKC8 cells grew inside a RGD-functionalized dextran hydrogel to obtain 3D spheroids. 13 days post-seeding, the spheroids were incubated with 35mM of D-glucose (HG), 5mM of D-glucose (LG) or 35mM of mannitol as osmotic control for 6h, 24h or 72h. The quality of the established 3D cell culture of mature HKC-8 spheroids was assessed by Aquoporin-1 and Glut-1 staining. After incubations quantitative-PCR analyses were performed for fibrotic and inflammatory markers. Immunofluorescence for fibrotic markers was performed on HKC-8 spheroids incubated for 72h. Results High glucose (HG) medium induced CCL5 gene expression on wild-type HKC-8 spheroids after 6h and 24h of incubation in comparison with the control group. Interestingly, in the ADAM17-deleted spheroids, CCL5 gene expression maintained similar to control after 6h of incubation with HG medium and tended to decrease after 24h of incubation in comparison with the wild-type. Collagen IV gene expression was increased in the wild-type spheroids incubated with HG in comparison with the control group. In ADAM17-deleted spheroids, Collagen IV gene expression was significantly decreased in the cells incubated with HG in comparison with the wild-type cells incubated with HG. HG increased the expression of α-SMA, fibronectin and Collagen IV in wild-type spheroids. Adam17 deletion blocked the increase of α-SMA, fibronectin and Collagen IV expression compared with wild-type cells after 72h of incubation. Conclusion ADAM17 blockade protects against fibrosis and inflammation in human kidney tubular spheroids under high glucose.

scholarly journals CXCR4 Promotes Renal Tubular Cell Survival in Male Diabetic Rats: Implications for Ligand Inactivation in the Human Kidney

Endocrinology ◽  
2015 ◽  
Vol 156 (3) ◽  
pp. 1121-1132 ◽  
Author(s):  
Ferhan S. Siddiqi ◽  
Li-Hao Chen ◽  
Suzanne L. Advani ◽  
Kerri Thai ◽  
Sri N. Batchu ◽  
...  
Keyword(s):  
Diabetic Nephropathy ◽  
Cell Survival ◽  
Cultured Cells ◽  
Human Kidney ◽  
Diabetic Rats ◽  
Cxcr4 Expression ◽  
Receptor Expression ◽  
Tubular Cell ◽  
Renal Tubular Cell ◽  
Tubulointerstitial Injury

Abstract Binding of the receptor CXCR4 to its ligand stromal cell–derived factor 1 (SDF-1) promotes cell survival and is under the influence of a number of regulatory processes including enzymatic ligand inactivation by endopeptidases such as matrix metalloproteinase 9 (MMP-9). In light of the pivotal role that the SDF-1/CXCR4 axis plays in renal development and in the pathological growth of renal cells, we explored the function of this pathway in diabetic rats and in biopsies from patients with diabetic nephropathy, hypothesizing that the pro-survival effects of CXCR4 in resident cells would attenuate renal injury. Renal CXCR4 expression was observed to be increased in diabetic rats, whereas antagonism of the receptor unmasked albuminuria and accelerated tubular epithelial cell death. In cultured cells, CXCR4 blockade promoted tubular cell apoptosis, up-regulated Bcl-2-associated death promoter, and prevented high glucose/SDF-1-augmented phosphorylation of the pro-survival kinase, Akt. Although CXCR4 expression was also increased in biopsy tissue from patients with diabetic nephropathy, serine 339 phosphorylation of the receptor, indicative of ligand engagement, was unaffected. Coincident with these changes in receptor expression but not activity, MMP-9 was also up-regulated in diabetic nephropathy biopsies. Supporting a ligand-inactivating effect of the endopeptidase, exposure of cultured cells to recombinant MMP-9 abrogated SDF-1 induced Akt phosphorylation. These observations demonstrate a potentially reno-protective role for CXCR4 in diabetes that is impeded in its actions in the human kidney by the coincident up-regulation of ligand-inactivating endopeptidases. Therapeutically intervening in this interplay may limit tubulointerstitial injury, the principal determinant of renal decline in diabetes.

scholarly journals AEBP1 is Upregulated in Diabetic Nephropathy Biopsies

2021 ◽  
Vol 4 (1) ◽  
Author(s):  
Benjamin Freije ◽  
Ricardo Melo Ferreira ◽  
Ying-Hua Cheng ◽  
Samir Parikh ◽  
Michael Eadon
Keyword(s):  
Diabetic Nephropathy ◽  
Kidney Disease ◽  
Interstitial Fibrosis ◽  
Treatment Strategies ◽  
Human Kidney ◽  
Sequencing Data ◽  
Specific Expression ◽  
Enhancer Binding Protein ◽  
Organ Specific ◽  
Matrix Organization

Background: Worldwide, one in eleven adults have diabetes mellitus and 30% to 40% will develop diabetic kidney disease (DKD). A mechanistic understanding of DKD is crucial to develop treatment strategies. To unravel DKD’s pathogenesis, single cell (scRNA) sequencing has proven a powerful tool, but is limited by a lack of localization. Spatial transcriptomics allows the mapping of scRNA sequencing data back to histology. Methods: Frozen human nephrectomy and biopsy samples were processed according to Visium spatial gene expression protocols, stained with H&E, and imaged. Samples were permeabilized for RNA capture, reverse transcribed and sequenced on an Illumina NovaSeq 6000. Mapping and counting were completed in Space Ranger and data was processed in Seurat. Samples were laser microdissected, protein was isolated, and protein was quantified by HPLC-MS. Results: Clusters from scRNAseq were mapped upon reference and DKD spatial transcriptomic images (N=4 reference, 2 DKD). Differentially expressed genes were identified in diabetic kidneys, including the upregulation of Adipocyte Enhancer Binding Protein (AEBP1).  Pathway analysis revealed enrichment of extracellular matrix organization and immune process pathways. To increase the confidence of these findings, glomeruli and the tubulointerstitium were laser microdissected (N=7 diseased, 4 reference) for proteomic analysis. AEBP1 was upregulated in the tubular interstitium of diseased kidneys and selectively upregulated in the glomeruli of Diabetic Nephropathy samples (N=2). AEBP1 localized to the interstitium by spatial transcriptomics and was expressed in highly fibrotic regions. Glomerular expression was not observed due to glomerulosclerosis. Conclusion: AEBP1 upregulation is a marker of interstitial fibrosis, with specific expression in the glomeruli of diabetic nephropathy specimens with glomerulosclerosis. Impact: This is the first study utilizing spatial transcriptomics to define and localize markers of human kidney disease. Confirmatory studies are required in larger sample sizes. AEBP1 is a previously unidentified marker of DKD previously associated with fibrosis in other organ-specific diseases.

scholarly journals Autophagy in diabetic nephropathy

2014 ◽  
Vol 224 (1) ◽  
pp. R15-R30 ◽  
Author(s):  
Yan Ding ◽  
Mary E Choi
Keyword(s):  
Type 2 Diabetes ◽  
Diabetic Nephropathy ◽  
Transforming Growth Factor ◽  
Renin Angiotensin System ◽  
Human Kidney ◽  
Nutrient Sensing ◽  
Signal Pathways ◽  
End Stage

Diabetic nephropathy (DN) is the most common cause of end-stage kidney disease worldwide, and is associated with increased morbidity and mortality in patients with both type 1 and type 2 diabetes. Increasing prevalence of diabetes has made the need for effective treatment of DN critical and thereby identifying new therapeutic targets to improve clinical management. Autophagy is a highly conserved ‘self-eating’ pathway by which cells degrade and recycle macromolecules and organelles. Autophagy serves as an essential mechanism to maintain homeostasis of glomeruli and tubules, and plays important roles in human health and diseases. Impairment of autophagy is implicated in the pathogenesis of DN. Emerging body of evidence suggests that targeting the autophagic pathway to activate and restore autophagy activity may be renoprotective. In this review, we examine current advances in our understanding of the roles of autophagy in diabetic kidney injury, focusing on studies in renal cells in culture, human kidney tissues, and experimental animal models of diabetes. We discuss the major nutrient-sensing signal pathways and diabetes-induced altered intracellular metabolism and cellular events, including accumulation of advanced glycation end-products, increased oxidative stress, endoplasmic reticulum stress, hypoxia, and activation of the renin–angiotensin system, which modulate autophagic activity and contribute to the development of DN. We also highlight recent studies of autophagy and transforming growth factor-β in renal fibrosis, the final common response to injury that ultimately leads to end-stage kidney failure in both type 1 and type 2 diabetes. These findings suggest the possibility that autophagy can be a therapeutic target against DN.

scholarly journals A receptor for the third component of complement in the human renal glomerulus

1975 ◽  
Vol 142 (4) ◽  
pp. 1029-1034 ◽  
Author(s):  
MC Gelfand ◽  
MM Frank ◽  
I Green
Keyword(s):  
Human Kidney ◽  
Complement C3 ◽  
Renal Diseases ◽  
Renal Glomerulus ◽  
Sheep Erythrocytes ◽  
Tissue Sections ◽  
The Third ◽  
Third Component Of Complement

In the course of studying the nature of mononuclear cellular infiltrates in tissue sections of human kidney it was noted that indicator sheep erythrocytes densely coated with the third component of complement (C3) specifically adhered to all of the glomeruli in the tissue sections. The deposition of complement (C) within the glomerulus is a feature of many immunologically related renal diseases (1,2), yet the precise mechanism by which C is deposited remains unexplained. We feel that this observation, suggesting the presence of a receptor for C, is, therefore, of particular interest.

scholarly journals Endoglin Promotes Myofibroblast Differentiation and Extracellular Matrix Production in Diabetic Nephropathy

2020 ◽  
Vol 21 (20) ◽  
pp. 7713
Author(s):  
Tessa Gerrits ◽  
Malu Zandbergen ◽  
Ron Wolterbeek ◽  
Jan A. Bruijn ◽  
Hans J. Baelde ◽  
...  
Keyword(s):  
Diabetic Nephropathy ◽  
Cell Function ◽  
Interstitial Fibrosis ◽  
Human Kidney ◽  
Fibroblast Cell ◽  
Diabetic Patients ◽  
Endothelial Cell Function ◽  
Intestinal Fibrosis ◽  
Kidney Fibroblast ◽  
Sirius Red

Diabetic nephropathy (DN) is a complication of diabetes mellitus that can lead to proteinuria and a progressive decline in renal function. Endoglin, a co-receptor of TGF-β, is known primarily for regulating endothelial cell function; however, endoglin is also associated with hepatic, cardiac, and intestinal fibrosis. This study investigates whether endoglin contributes to the development of interstitial fibrosis in DN. Kidney autopsy material from 80 diabetic patients was stained for endoglin and Sirius Red and scored semi-quantitatively. Interstitial endoglin expression was increased in samples with DN and was correlated with Sirius Red staining (p < 0.001). Endoglin expression was also correlated with reduced eGFR (p = 0.001), increased creatinine (p < 0.01), increased systolic blood pressure (p < 0.05), hypertension (p < 0.05), and higher IFTA scores (p < 0.001). Biopsy samples from DN patients were also co-immunostained for endoglin together with CD31, CD68, vimentin, or α-SMA Endoglin co-localized with both the endothelial marker CD31 and the myofibroblast marker α-SMA. Finally, we used shRNA to knockdown endoglin expression in a human kidney fibroblast cell line. We found that TGF-β1 stimulation upregulated SERPINE1, CTGF, and ACTA2 mRNA and α-SMA protein, and that these effects were significantly reduced in fibroblasts after endoglin knockdown. Taken together, these data suggest that endoglin plays a role in the pathogenesis of interstitial fibrosis in DN.

EXPRESSION OF CYCLOOXYGENASE-2 IN THE MACULA DENSA OF HUMAN KIDNEY IN HYPERTENSION, CONGESTIVE HEART FAILURE, AND DIABETIC NEPHROPATHY

Renal Failure ◽  
2001 ◽  
Vol 23 (3-4) ◽  
pp. 321-330 ◽  
Author(s):  
Kanwar Nasir M. Khan ◽  
Allen Burke ◽  
Kristina M. Stanfield ◽  
Richard K. Harris ◽  
David A. Baron
Keyword(s):  
Heart Failure ◽  
Congestive Heart Failure ◽  
Diabetic Nephropathy ◽  
Human Kidney ◽  
Cyclooxygenase 2 ◽  
Macula Densa

scholarly journals Glomerular Function and Structural Integrity Depend on Hyaluronan Synthesis by Glomerular Endothelium

2019 ◽  
Vol 30 (10) ◽  
pp. 1886-1897 ◽  
Author(s):  
Bernard M. van den Berg ◽  
Gangqi Wang ◽  
Margien G. S. Boels ◽  
M. Cristina Avramut ◽  
Erik Jansen ◽  
...  
Keyword(s):  
Diabetic Nephropathy ◽  
Barrier Function ◽  
Structural Integrity ◽  
Specific Binding ◽  
Kidney Tissue ◽  
Human Kidney ◽  
Endothelial Glycocalyx ◽  
Adult Mice ◽  
Filtration Barrier ◽  
And Function

BackgroundA glycocalyx envelope consisting of proteoglycans and adhering proteins covers endothelial cells, both the luminal and abluminal surface. We previously demonstrated that short-term loss of integrity of the luminal glycocalyx layer resulted in perturbed glomerular filtration barrier function.MethodsTo explore the role of the glycocalyx layer of the endothelial extracellular matrix in renal function, we generated mice with an endothelium-specific and inducible deletion of hyaluronan synthase 2 (Has2), the enzyme that produces hyaluronan, the main structural component of the endothelial glycocalyx layer. We also investigated the presence of endothelial hyaluronan in human kidney tissue from patients with varying degrees of diabetic nephropathy.ResultsEndothelial deletion of Has2 in adult mice led to substantial loss of the glycocalyx structure, and analysis of their kidneys and kidney function showed vascular destabilization, characterized by mesangiolysis, capillary ballooning, and albuminuria. This process develops over time into glomerular capillary rarefaction and glomerulosclerosis, recapitulating the phenotype of progressive human diabetic nephropathy. Using a hyaluronan-specific probe, we found loss of glomerular endothelial hyaluronan in association with lesion formation in tissue from patients with diabetic nephropathy. We also demonstrated that loss of hyaluronan, which harbors a specific binding site for angiopoietin and a key regulator of endothelial quiescence and maintenance of EC barrier function results in disturbed angiopoietin 1 Tie2.ConclusionsEndothelial loss of hyaluronan results in disturbed glomerular endothelial stabilization. Glomerular endothelial hyaluronan is a previously unrecognized key component of the extracelluar matrix that is required for glomerular structure and function and lost in diabetic nephropathy.

scholarly journals Mice deficient in PAPP-A show resistance to the development of diabetic nephropathy

2013 ◽  
Vol 219 (1) ◽  
pp. 51-58 ◽  
Author(s):  
Jessica R Mader ◽  
Zachary T Resch ◽  
Gary R McLean ◽  
Jakob H Mikkelsen ◽  
Claus Oxvig ◽  
...  
Keyword(s):  
Diabetic Nephropathy ◽  
Mononuclear Cell ◽  
Human Kidney ◽  
Cell Infiltration ◽  
Mononuclear Cell Infiltration ◽  
Specific Staining ◽  
Age Related ◽  
Mesangial Area ◽  
Glomerular Size ◽  
Bowman's Capsule

We investigated pregnancy-associated plasma protein-A (PAPP-A) in diabetic nephropathy. Normal human kidney showed specific staining for PAPP-A in glomeruli, and this staining was markedly increased in diabetic kidney. To assess the possible contribution of PAPP-A in the development of diabetic nephropathy, we induced diabetes with streptozotocin in 14-month-old WT and Papp-A knockout (KO) mice. Renal histopathology was evaluated after 4 months of stable hyperglycemia. Kidneys from diabetic WT mice showed multiple abnormalities including thickening of Bowman's capsule (100% of mice), increased glomerular size (80% of mice), tubule dilation (80% of mice), and mononuclear cell infiltration (90% of mice). Kidneys of age-matched non-diabetic WT mice had similar evidence of tubule dilation and mononuclear cell infiltration to those of diabetic WT mice, indicating that these changes were predominantly age-related. However, thickened Bowman's capsule and increased glomerular size appeared specific for the experimental diabetes. Kidneys from diabetic Papp-A KO mice had significantly reduced or no evidence of changes in Bowman's capsule thickening and glomerular size. There was also a shift to larger mesangial area and increased macrophage staining in diabetic WT mice compared with Papp-A KO mice. In summary, elevated PAPP-A expression in glomeruli is associated with diabetic nephropathy in humans and absence of PAPP-A is associated with resistance to the development of indicators of diabetic nephropathy in mice. These data suggest PAPP-A as a potential therapeutic target for diabetic nephropathy.

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