Perturbation of ATP6V0A1 as a driver and druggable target for renal proximal tubular cellular injury in nephropathic cystinosis

Understanding the unique susceptibility of the human kidney to pH dysfunction and injury in cystinosis is paramount to develop new therapies to preserve renal function. Renal proximal tubular epithelial cells (RPTECs) and fibroblasts, isolated from patients with cystinosis were transcriptionally pro...

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Autores: Sur, Swastika, Kerwin, Maggie, Pineda Sanjuan, Silvia, Sansanwal, Poonam, Sigdel, Tara K., Sirota, Marina, Sarwal, Minnie M.
Tipo de recurso: artículo
Fecha de publicación:2022
País:España
Institución:Universidad Complutense de Madrid (UCM)
Repositorio:Docta Complutense
Idioma:inglés
OAI Identifier:oai:docta.ucm.es:20.500.14352/102622
Acceso en línea:https://hdl.handle.net/20.500.14352/102622
Access Level:acceso abierto
Palabra clave:611.61
616.61
57.087.1
519.22-76
Cystinosis
kidney disease ESRD,
Genetic renal
ESRD
Autophagy
CKD
Nefrología y urología
Genética médica
Estadística
3205.06 Nefrología
2404.01 Bioestadística
3205 Medicina Interna
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oai_identifier_str oai:docta.ucm.es:20.500.14352/102622
network_acronym_str ES
network_name_str España
repository_id_str
spelling Perturbation of ATP6V0A1 as a driver and druggable target for renal proximal tubular cellular injury in nephropathic cystinosisSur, SwastikaKerwin, MaggiePineda Sanjuan, SilviaSansanwal, PoonamSigdel, Tara K.Sirota, MarinaSarwal, Minnie M.611.61616.6157.087.1519.22-76Cystinosiskidney disease ESRD,Genetic renalESRDAutophagyCKDNefrología y urologíaGenética médicaEstadística3205.06 Nefrología2404.01 Bioestadística3205 Medicina InternaUnderstanding the unique susceptibility of the human kidney to pH dysfunction and injury in cystinosis is paramount to develop new therapies to preserve renal function. Renal proximal tubular epithelial cells (RPTECs) and fibroblasts, isolated from patients with cystinosis were transcriptionally profiled. Lysosomal fractionation, immunoblotting, confocal microscopy, intracellular pH, TEM, mitochondrial stress test, and membrane integrity assays were performed for validation. CRISPR, CTNS-/- RPTECs were generated. A new compound, Astaxanthin (ATX), was evaluated for rescue of the CTNS -/- phenotype. Alterations in cell stress, pH, autophagic turnover, and mitochondrial energetics, highlighted key changes in the vacuolar (V)-ATPases in patient derived and CTNS-/- RPTECs. ATP6V0A1 was significantly downregulated in cystinosis and highly co-regulated with loss of CTNS. Overexpression of ATP6V0A1 rescued cell stress and mitochondrial function. Treatment of CTNS -/- RPTECs with ATX, induced ATP6V0A1 expression and the resulting rescue of the RPTE cell and mitochondrial injury. In conclusion, our exploratory transcriptional and in vitro cellular and functional studies confirm that loss of cystinosin in RPTEC, results in a reduction in ATP6V0A1 expression, with changes in intracellular pH, mitochondrial integrity, mitochondrial function, and autophagosome-lysosome clearance. ATX can rescue the cystinotic RPTEC injury, at least partially mediated by upregulating ATP6V0A1 expression. The availability of ATX as a well-tolerated oral supplement, offers its further clinical evaluation as a potential novel therapeutic to limit renal tubular injury in cystinosis, independent of cysteine depletion alone. Competing Interest Statement The authors have declared no competing interest.Universidad Complutense de Madrid20222022-05-1320222022-05-13journal articlehttp://purl.org/coar/resource_type/c_6501AOhttp://purl.org/coar/version/c_b1a7d7d4d402bcceinfo:eu-repo/semantics/articleapplication/pdfhttps://hdl.handle.net/20.500.14352/102622reponame:Docta Complutenseinstname:Universidad Complutense de Madrid (UCM)Inglésengopen accesshttp://purl.org/coar/access_right/c_abf2Attribution-NonCommercial-NoDerivatives 4.0 Internationalhttp://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessoai:docta.ucm.es:20.500.14352/1026222026-06-02T12:44:21Z
dc.title.none.fl_str_mv Perturbation of ATP6V0A1 as a driver and druggable target for renal proximal tubular cellular injury in nephropathic cystinosis
title Perturbation of ATP6V0A1 as a driver and druggable target for renal proximal tubular cellular injury in nephropathic cystinosis
spellingShingle Perturbation of ATP6V0A1 as a driver and druggable target for renal proximal tubular cellular injury in nephropathic cystinosis
Sur, Swastika
611.61
616.61
57.087.1
519.22-76
Cystinosis
kidney disease ESRD,
Genetic renal
ESRD
Autophagy
CKD
Nefrología y urología
Genética médica
Estadística
3205.06 Nefrología
2404.01 Bioestadística
3205 Medicina Interna
title_short Perturbation of ATP6V0A1 as a driver and druggable target for renal proximal tubular cellular injury in nephropathic cystinosis
title_full Perturbation of ATP6V0A1 as a driver and druggable target for renal proximal tubular cellular injury in nephropathic cystinosis
title_fullStr Perturbation of ATP6V0A1 as a driver and druggable target for renal proximal tubular cellular injury in nephropathic cystinosis
title_full_unstemmed Perturbation of ATP6V0A1 as a driver and druggable target for renal proximal tubular cellular injury in nephropathic cystinosis
title_sort Perturbation of ATP6V0A1 as a driver and druggable target for renal proximal tubular cellular injury in nephropathic cystinosis
dc.creator.none.fl_str_mv Sur, Swastika
Kerwin, Maggie
Pineda Sanjuan, Silvia
Sansanwal, Poonam
Sigdel, Tara K.
Sirota, Marina
Sarwal, Minnie M.
author Sur, Swastika
author_facet Sur, Swastika
Kerwin, Maggie
Pineda Sanjuan, Silvia
Sansanwal, Poonam
Sigdel, Tara K.
Sirota, Marina
Sarwal, Minnie M.
author_role author
author2 Kerwin, Maggie
Pineda Sanjuan, Silvia
Sansanwal, Poonam
Sigdel, Tara K.
Sirota, Marina
Sarwal, Minnie M.
author2_role author
author
author
author
author
author
dc.contributor.none.fl_str_mv Universidad Complutense de Madrid
dc.subject.none.fl_str_mv 611.61
616.61
57.087.1
519.22-76
Cystinosis
kidney disease ESRD,
Genetic renal
ESRD
Autophagy
CKD
Nefrología y urología
Genética médica
Estadística
3205.06 Nefrología
2404.01 Bioestadística
3205 Medicina Interna
topic 611.61
616.61
57.087.1
519.22-76
Cystinosis
kidney disease ESRD,
Genetic renal
ESRD
Autophagy
CKD
Nefrología y urología
Genética médica
Estadística
3205.06 Nefrología
2404.01 Bioestadística
3205 Medicina Interna
description Understanding the unique susceptibility of the human kidney to pH dysfunction and injury in cystinosis is paramount to develop new therapies to preserve renal function. Renal proximal tubular epithelial cells (RPTECs) and fibroblasts, isolated from patients with cystinosis were transcriptionally profiled. Lysosomal fractionation, immunoblotting, confocal microscopy, intracellular pH, TEM, mitochondrial stress test, and membrane integrity assays were performed for validation. CRISPR, CTNS-/- RPTECs were generated. A new compound, Astaxanthin (ATX), was evaluated for rescue of the CTNS -/- phenotype. Alterations in cell stress, pH, autophagic turnover, and mitochondrial energetics, highlighted key changes in the vacuolar (V)-ATPases in patient derived and CTNS-/- RPTECs. ATP6V0A1 was significantly downregulated in cystinosis and highly co-regulated with loss of CTNS. Overexpression of ATP6V0A1 rescued cell stress and mitochondrial function. Treatment of CTNS -/- RPTECs with ATX, induced ATP6V0A1 expression and the resulting rescue of the RPTE cell and mitochondrial injury. In conclusion, our exploratory transcriptional and in vitro cellular and functional studies confirm that loss of cystinosin in RPTEC, results in a reduction in ATP6V0A1 expression, with changes in intracellular pH, mitochondrial integrity, mitochondrial function, and autophagosome-lysosome clearance. ATX can rescue the cystinotic RPTEC injury, at least partially mediated by upregulating ATP6V0A1 expression. The availability of ATX as a well-tolerated oral supplement, offers its further clinical evaluation as a potential novel therapeutic to limit renal tubular injury in cystinosis, independent of cysteine depletion alone. Competing Interest Statement The authors have declared no competing interest.
publishDate 2022
dc.date.none.fl_str_mv 2022
2022-05-13
2022
2022-05-13
dc.type.none.fl_str_mv journal article
http://purl.org/coar/resource_type/c_6501
AO
http://purl.org/coar/version/c_b1a7d7d4d402bcce
dc.type.openaire.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv https://hdl.handle.net/20.500.14352/102622
url https://hdl.handle.net/20.500.14352/102622
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
Attribution-NonCommercial-NoDerivatives 4.0 International
http://creativecommons.org/licenses/by-nc-nd/4.0/
dc.rights.openaire.fl_str_mv info:eu-repo/semantics/openAccess
rights_invalid_str_mv open access
http://purl.org/coar/access_right/c_abf2
Attribution-NonCommercial-NoDerivatives 4.0 International
http://creativecommons.org/licenses/by-nc-nd/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.source.none.fl_str_mv reponame:Docta Complutense
instname:Universidad Complutense de Madrid (UCM)
instname_str Universidad Complutense de Madrid (UCM)
reponame_str Docta Complutense
collection Docta Complutense
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,300724

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