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Molecular and functional characterization of urine‐derived podocytes from patients with Alport syndrome
Alport syndrome (AS) is a genetic disorder involving mutations in the genes encoding collagen IV α3, α4 or α5 chains, resulting in the impairment of glomerular basement membrane. Podocytes are responsible for production and correct assembly of collagen IV isoforms; however, data on the phenotypic ch...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley & Sons, Ltd
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7589231/ https://www.ncbi.nlm.nih.gov/pubmed/32652570 http://dx.doi.org/10.1002/path.5496 |
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author | Iampietro, Corinne Bellucci, Linda Arcolino, Fanny O Arigoni, Maddalena Alessandri, Luca Gomez, Yonathan Papadimitriou, Elli Calogero, Raffaele A Cocchi, Enrico Van Den Heuvel, Lambertus Levtchenko, Elena Bussolati, Benedetta |
author_facet | Iampietro, Corinne Bellucci, Linda Arcolino, Fanny O Arigoni, Maddalena Alessandri, Luca Gomez, Yonathan Papadimitriou, Elli Calogero, Raffaele A Cocchi, Enrico Van Den Heuvel, Lambertus Levtchenko, Elena Bussolati, Benedetta |
author_sort | Iampietro, Corinne |
collection | PubMed |
description | Alport syndrome (AS) is a genetic disorder involving mutations in the genes encoding collagen IV α3, α4 or α5 chains, resulting in the impairment of glomerular basement membrane. Podocytes are responsible for production and correct assembly of collagen IV isoforms; however, data on the phenotypic characteristics of human AS podocytes and their functional alterations are currently limited. The evident loss of viable podocytes into the urine of patients with active glomerular disease enables their isolation in a non‐invasive way. Here we isolated, immortalized, and subcloned podocytes from the urine of three different AS patients for molecular and functional characterization. AS podocytes expressed a typical podocyte signature and showed a collagen IV profile reflecting each patient's mutation. Furthermore, RNA‐sequencing analysis revealed 348 genes differentially expressed in AS podocytes compared with control podocytes. Gene Ontology analysis underlined the enrichment in genes involved in cell motility, adhesion, survival, and angiogenesis. In parallel, AS podocytes displayed reduced motility. Finally, a functional permeability assay, using a podocyte–glomerular endothelial cell co‐culture system, was established and AS podocyte co‐cultures showed a significantly higher permeability of albumin compared to control podocyte co‐cultures, in both static and dynamic conditions under continuous perfusion. In conclusion, our data provide a molecular characterization of immortalized AS podocytes, highlighting alterations in several biological processes related to extracellular matrix remodelling. Moreover, we have established an in vitro model to reproduce the altered podocyte permeability observed in patients with AS. © 2020 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of Pathological Society of Great Britain and Ireland.. |
format | Online Article Text |
id | pubmed-7589231 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | John Wiley & Sons, Ltd |
record_format | MEDLINE/PubMed |
spelling | pubmed-75892312020-10-30 Molecular and functional characterization of urine‐derived podocytes from patients with Alport syndrome Iampietro, Corinne Bellucci, Linda Arcolino, Fanny O Arigoni, Maddalena Alessandri, Luca Gomez, Yonathan Papadimitriou, Elli Calogero, Raffaele A Cocchi, Enrico Van Den Heuvel, Lambertus Levtchenko, Elena Bussolati, Benedetta J Pathol Original Papers Alport syndrome (AS) is a genetic disorder involving mutations in the genes encoding collagen IV α3, α4 or α5 chains, resulting in the impairment of glomerular basement membrane. Podocytes are responsible for production and correct assembly of collagen IV isoforms; however, data on the phenotypic characteristics of human AS podocytes and their functional alterations are currently limited. The evident loss of viable podocytes into the urine of patients with active glomerular disease enables their isolation in a non‐invasive way. Here we isolated, immortalized, and subcloned podocytes from the urine of three different AS patients for molecular and functional characterization. AS podocytes expressed a typical podocyte signature and showed a collagen IV profile reflecting each patient's mutation. Furthermore, RNA‐sequencing analysis revealed 348 genes differentially expressed in AS podocytes compared with control podocytes. Gene Ontology analysis underlined the enrichment in genes involved in cell motility, adhesion, survival, and angiogenesis. In parallel, AS podocytes displayed reduced motility. Finally, a functional permeability assay, using a podocyte–glomerular endothelial cell co‐culture system, was established and AS podocyte co‐cultures showed a significantly higher permeability of albumin compared to control podocyte co‐cultures, in both static and dynamic conditions under continuous perfusion. In conclusion, our data provide a molecular characterization of immortalized AS podocytes, highlighting alterations in several biological processes related to extracellular matrix remodelling. Moreover, we have established an in vitro model to reproduce the altered podocyte permeability observed in patients with AS. © 2020 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of Pathological Society of Great Britain and Ireland.. John Wiley & Sons, Ltd 2020-08-19 2020-09 /pmc/articles/PMC7589231/ /pubmed/32652570 http://dx.doi.org/10.1002/path.5496 Text en © 2020 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of Pathological Society of Great Britain and Ireland. This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Original Papers Iampietro, Corinne Bellucci, Linda Arcolino, Fanny O Arigoni, Maddalena Alessandri, Luca Gomez, Yonathan Papadimitriou, Elli Calogero, Raffaele A Cocchi, Enrico Van Den Heuvel, Lambertus Levtchenko, Elena Bussolati, Benedetta Molecular and functional characterization of urine‐derived podocytes from patients with Alport syndrome |
title | Molecular and functional characterization of urine‐derived podocytes from patients with Alport syndrome |
title_full | Molecular and functional characterization of urine‐derived podocytes from patients with Alport syndrome |
title_fullStr | Molecular and functional characterization of urine‐derived podocytes from patients with Alport syndrome |
title_full_unstemmed | Molecular and functional characterization of urine‐derived podocytes from patients with Alport syndrome |
title_short | Molecular and functional characterization of urine‐derived podocytes from patients with Alport syndrome |
title_sort | molecular and functional characterization of urine‐derived podocytes from patients with alport syndrome |
topic | Original Papers |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7589231/ https://www.ncbi.nlm.nih.gov/pubmed/32652570 http://dx.doi.org/10.1002/path.5496 |
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