Cell-Based Phenotypic Drug Screening Identifies Luteolin as Candidate Therapeutic for Nephropathic Cystinosis.
Amino Acid Transport Systems, Neutral
/ genetics
Animals
Antioxidants
/ adverse effects
Apoptosis
/ drug effects
Autophagy
/ drug effects
Cells, Cultured
Cystinosis
/ drug therapy
Disease Models, Animal
Drug Evaluation, Preclinical
/ methods
Endocytosis
/ drug effects
Humans
Kidney Tubules, Proximal
/ pathology
Low Density Lipoprotein Receptor-Related Protein-2
/ metabolism
Luteolin
/ adverse effects
Lysosomes
/ drug effects
Mice
Oxidative Stress
/ drug effects
Phenotype
RNA, Messenger
/ metabolism
Sequestosome-1 Protein
/ genetics
Zebrafish
apoptosis
autophagy
endocytosis
lysosomal storage disease
reactive
Journal
Journal of the American Society of Nephrology : JASN
ISSN: 1533-3450
Titre abrégé: J Am Soc Nephrol
Pays: United States
ID NLM: 9013836
Informations de publication
Date de publication:
07 2020
07 2020
Historique:
received:
23
09
2019
accepted:
04
04
2020
pubmed:
7
6
2020
medline:
25
2
2021
entrez:
7
6
2020
Statut:
ppublish
Résumé
Mutations in the gene that encodes the lysosomal cystine transporter cystinosin cause the lysosomal storage disease cystinosis. Defective cystine transport leads to intralysosomal accumulation and crystallization of cystine. The most severe phenotype, nephropathic cystinosis, manifests during the first months of life, as renal Fanconi syndrome. The cystine-depleting agent cysteamine significantly delays symptoms, but it cannot prevent progression to ESKD and does not treat Fanconi syndrome. This suggests the involvement of pathways in nephropathic cystinosis that are unrelated to lysosomal cystine accumulation. Recent data indicate that one such potential pathway, lysosome-mediated degradation of autophagy cargoes, is compromised in cystinosis. To identify drugs that reduce levels of the autophagy-related protein p62/SQSTM1 in cystinotic proximal tubular epithelial cells, we performed a high-throughput screening on the basis of an in-cell ELISA assay. We then tested a promising candidate in cells derived from patients with, and mouse models of, cystinosis, and in preclinical studies in cystinotic zebrafish. Of 46 compounds identified as reducing p62/SQSTM1 levels in cystinotic cells, we selected luteolin on the basis of its efficacy, safety profile, and similarity to genistein, which we previously showed to ameliorate other lysosomal abnormalities of cystinotic cells. Our data show that luteolin improves the autophagy-lysosome degradative pathway, is a powerful antioxidant, and has antiapoptotic properties. Moreover, luteolin stimulates endocytosis and improves the expression of the endocytic receptor megalin. Our data show that luteolin improves defective pathways of cystinosis and has a good safety profile, and thus has potential as a treatment for nephropathic cystinosis and other renal lysosomal storage diseases.
Sections du résumé
BACKGROUND
Mutations in the gene that encodes the lysosomal cystine transporter cystinosin cause the lysosomal storage disease cystinosis. Defective cystine transport leads to intralysosomal accumulation and crystallization of cystine. The most severe phenotype, nephropathic cystinosis, manifests during the first months of life, as renal Fanconi syndrome. The cystine-depleting agent cysteamine significantly delays symptoms, but it cannot prevent progression to ESKD and does not treat Fanconi syndrome. This suggests the involvement of pathways in nephropathic cystinosis that are unrelated to lysosomal cystine accumulation. Recent data indicate that one such potential pathway, lysosome-mediated degradation of autophagy cargoes, is compromised in cystinosis.
METHODS
To identify drugs that reduce levels of the autophagy-related protein p62/SQSTM1 in cystinotic proximal tubular epithelial cells, we performed a high-throughput screening on the basis of an in-cell ELISA assay. We then tested a promising candidate in cells derived from patients with, and mouse models of, cystinosis, and in preclinical studies in cystinotic zebrafish.
RESULTS
Of 46 compounds identified as reducing p62/SQSTM1 levels in cystinotic cells, we selected luteolin on the basis of its efficacy, safety profile, and similarity to genistein, which we previously showed to ameliorate other lysosomal abnormalities of cystinotic cells. Our data show that luteolin improves the autophagy-lysosome degradative pathway, is a powerful antioxidant, and has antiapoptotic properties. Moreover, luteolin stimulates endocytosis and improves the expression of the endocytic receptor megalin.
CONCLUSIONS
Our data show that luteolin improves defective pathways of cystinosis and has a good safety profile, and thus has potential as a treatment for nephropathic cystinosis and other renal lysosomal storage diseases.
Identifiants
pubmed: 32503896
pii: ASN.2019090956
doi: 10.1681/ASN.2019090956
pmc: PMC7351012
doi:
Substances chimiques
Amino Acid Transport Systems, Neutral
0
Antioxidants
0
CTNS protein, human
0
Low Density Lipoprotein Receptor-Related Protein-2
0
RNA, Messenger
0
SQSTM1 protein, human
0
Sequestosome-1 Protein
0
Sqstm1 protein, mouse
0
Luteolin
KUX1ZNC9J2
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
1522-1537Informations de copyright
Copyright © 2020 by the American Society of Nephrology.
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