Cargando…

Urinary Extracellular Vesicles for Renal Tubular Transporters Expression in Patients With Gitelman Syndrome

Background: The utility of urinary extracellular vesicles (uEVs) to faithfully represent the changes of renal tubular protein expression remains unclear. We aimed to evaluate renal tubular sodium (Na(+)) or potassium (K(+)) associated transporters expression from uEVs and kidney tissues in patients...

Descripción completa

Detalles Bibliográficos
Autores principales: Sung, Chih-Chien, Chen, Min-Hsiu, Lin, Yi-Chang, Lin, Yu-Chun, Lin, Yi-Jia, Yang, Sung-Sen, Lin, Shih-Hua
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8219937/
https://www.ncbi.nlm.nih.gov/pubmed/34179047
http://dx.doi.org/10.3389/fmed.2021.679171
_version_ 1783711048106246144
author Sung, Chih-Chien
Chen, Min-Hsiu
Lin, Yi-Chang
Lin, Yu-Chun
Lin, Yi-Jia
Yang, Sung-Sen
Lin, Shih-Hua
author_facet Sung, Chih-Chien
Chen, Min-Hsiu
Lin, Yi-Chang
Lin, Yu-Chun
Lin, Yi-Jia
Yang, Sung-Sen
Lin, Shih-Hua
author_sort Sung, Chih-Chien
collection PubMed
description Background: The utility of urinary extracellular vesicles (uEVs) to faithfully represent the changes of renal tubular protein expression remains unclear. We aimed to evaluate renal tubular sodium (Na(+)) or potassium (K(+)) associated transporters expression from uEVs and kidney tissues in patients with Gitelman syndrome (GS) caused by inactivating mutations in SLC12A3. Methods: uEVs were isolated by ultracentrifugation from 10 genetically-confirmed GS patients. Membrane transporters including Na(+)-hydrogen exchanger 3 (NHE3), Na(+)/K(+)/2Cl(−) cotransporter (NKCC2), NaCl cotransporter (NCC), phosphorylated NCC (p-NCC), epithelial Na(+) channel β (ENaCβ), pendrin, renal outer medullary K1 channel (ROMK), and large-conductance, voltage-activated and Ca(2+)-sensitive K(+) channel (Maxi-K) were examined by immunoblotting of uEVs and immunofluorescence of biopsied kidney tissues. Healthy and disease (bulimic patients) controls were also enrolled. Results: Characterization of uEVs was confirmed by nanoparticle tracking analysis, transmission electron microscopy, and immunoblotting. Compared with healthy controls, uEVs from GS patients showed NCC and p-NCC abundance were markedly attenuated but NHE3, ENaCβ, and pendrin abundance significantly increased. ROMK and Maxi-K abundance were also significantly accentuated. Immunofluorescence of the representative kidney tissues from GS patients also demonstrated the similar findings to uEVs. uEVs from bulimic patients showed an increased abundance of NCC and p-NCC as well as NHE3, NKCC2, ENaCβ, pendrin, ROMK and Maxi-K, akin to that in immunofluorescence of their kidney tissues. Conclusion: uEVs could be a non-invasive tool to diagnose and evaluate renal tubular transporter adaptation in patients with GS and may be applied to other renal tubular diseases.
format Online
Article
Text
id pubmed-8219937
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher Frontiers Media S.A.
record_format MEDLINE/PubMed
spelling pubmed-82199372021-06-24 Urinary Extracellular Vesicles for Renal Tubular Transporters Expression in Patients With Gitelman Syndrome Sung, Chih-Chien Chen, Min-Hsiu Lin, Yi-Chang Lin, Yu-Chun Lin, Yi-Jia Yang, Sung-Sen Lin, Shih-Hua Front Med (Lausanne) Medicine Background: The utility of urinary extracellular vesicles (uEVs) to faithfully represent the changes of renal tubular protein expression remains unclear. We aimed to evaluate renal tubular sodium (Na(+)) or potassium (K(+)) associated transporters expression from uEVs and kidney tissues in patients with Gitelman syndrome (GS) caused by inactivating mutations in SLC12A3. Methods: uEVs were isolated by ultracentrifugation from 10 genetically-confirmed GS patients. Membrane transporters including Na(+)-hydrogen exchanger 3 (NHE3), Na(+)/K(+)/2Cl(−) cotransporter (NKCC2), NaCl cotransporter (NCC), phosphorylated NCC (p-NCC), epithelial Na(+) channel β (ENaCβ), pendrin, renal outer medullary K1 channel (ROMK), and large-conductance, voltage-activated and Ca(2+)-sensitive K(+) channel (Maxi-K) were examined by immunoblotting of uEVs and immunofluorescence of biopsied kidney tissues. Healthy and disease (bulimic patients) controls were also enrolled. Results: Characterization of uEVs was confirmed by nanoparticle tracking analysis, transmission electron microscopy, and immunoblotting. Compared with healthy controls, uEVs from GS patients showed NCC and p-NCC abundance were markedly attenuated but NHE3, ENaCβ, and pendrin abundance significantly increased. ROMK and Maxi-K abundance were also significantly accentuated. Immunofluorescence of the representative kidney tissues from GS patients also demonstrated the similar findings to uEVs. uEVs from bulimic patients showed an increased abundance of NCC and p-NCC as well as NHE3, NKCC2, ENaCβ, pendrin, ROMK and Maxi-K, akin to that in immunofluorescence of their kidney tissues. Conclusion: uEVs could be a non-invasive tool to diagnose and evaluate renal tubular transporter adaptation in patients with GS and may be applied to other renal tubular diseases. Frontiers Media S.A. 2021-06-09 /pmc/articles/PMC8219937/ /pubmed/34179047 http://dx.doi.org/10.3389/fmed.2021.679171 Text en Copyright © 2021 Sung, Chen, Lin, Lin, Lin, Yang and Lin. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Medicine
Sung, Chih-Chien
Chen, Min-Hsiu
Lin, Yi-Chang
Lin, Yu-Chun
Lin, Yi-Jia
Yang, Sung-Sen
Lin, Shih-Hua
Urinary Extracellular Vesicles for Renal Tubular Transporters Expression in Patients With Gitelman Syndrome
title Urinary Extracellular Vesicles for Renal Tubular Transporters Expression in Patients With Gitelman Syndrome
title_full Urinary Extracellular Vesicles for Renal Tubular Transporters Expression in Patients With Gitelman Syndrome
title_fullStr Urinary Extracellular Vesicles for Renal Tubular Transporters Expression in Patients With Gitelman Syndrome
title_full_unstemmed Urinary Extracellular Vesicles for Renal Tubular Transporters Expression in Patients With Gitelman Syndrome
title_short Urinary Extracellular Vesicles for Renal Tubular Transporters Expression in Patients With Gitelman Syndrome
title_sort urinary extracellular vesicles for renal tubular transporters expression in patients with gitelman syndrome
topic Medicine
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8219937/
https://www.ncbi.nlm.nih.gov/pubmed/34179047
http://dx.doi.org/10.3389/fmed.2021.679171
work_keys_str_mv AT sungchihchien urinaryextracellularvesiclesforrenaltubulartransportersexpressioninpatientswithgitelmansyndrome
AT chenminhsiu urinaryextracellularvesiclesforrenaltubulartransportersexpressioninpatientswithgitelmansyndrome
AT linyichang urinaryextracellularvesiclesforrenaltubulartransportersexpressioninpatientswithgitelmansyndrome
AT linyuchun urinaryextracellularvesiclesforrenaltubulartransportersexpressioninpatientswithgitelmansyndrome
AT linyijia urinaryextracellularvesiclesforrenaltubulartransportersexpressioninpatientswithgitelmansyndrome
AT yangsungsen urinaryextracellularvesiclesforrenaltubulartransportersexpressioninpatientswithgitelmansyndrome
AT linshihhua urinaryextracellularvesiclesforrenaltubulartransportersexpressioninpatientswithgitelmansyndrome