TY - JOUR
T1 - Cell-based phenotypic drug screening identifies luteolin as candidate therapeutic for nephropathic cystinosis
AU - de Leo, Ester
AU - Elmonem, Mohamed A.
AU - Berlingerio, Sante Princiero
AU - Berquez, Marine
AU - Festa, Beatrice Paola
AU - Raso, Roberto
AU - Bellomo, Francesco
AU - Starborg, Tobias
AU - Janssen, Manoe Jacoba
AU - Abbaszadeh, Zeinab
AU - Cairoli, Sara
AU - Goffredo, Bianca Maria
AU - Masereeuw, Rosalinde
AU - Devuyst, Olivier
AU - Lowe, Martin
AU - Levtchenko, Elena
AU - Luciani, Alessandro
AU - Emma, Francesco
AU - Rega, Laura Rita
N1 - Funding Information: These studies were supported by Cystinosis Research Foundation grant CRF-2016-004 (to F. Emma), ERA-Net for Research Programmes on Rare Diseases grant E-Rare2-JTC2014 (to F. Emma, E. Levtchenko and R. Masereeuw), Zon-MW grant 113301402 (to R. Masereeuw), Fonds Wetenschappelijk Onderzoek Vlaanderen grant 1801110N (to E. Levtchenko), European Reference Network for Rare Kidney Diseases (ERKNet) grant 739532, Cystinosis Research Foundation grant CRFS-2017-007, and Swiss National Science Foundation grant 310030_146490 (to O. Devuyst). Publisher Copyright: Copyright © 2020 by the American Society of Nephrology
PY - 2020/7
Y1 - 2020/7
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85087465943&partnerID=8YFLogxK
U2 - https://doi.org/10.1681/ASN.2019090956
DO - https://doi.org/10.1681/ASN.2019090956
M3 - Article
C2 - 32503896
SN - 1046-6673
VL - 31
SP - 1522
EP - 1537
JO - Journal of the American Society of Nephrology
JF - Journal of the American Society of Nephrology
IS - 7
ER -