• Home
  • Search Results
  • Impaired autophagy bridges lysosomal storage disease and epithelial dysfunction in the kidney.

Impaired autophagy bridges lysosomal storage disease and epithelial dysfunction in the kidney.

Nature communications (2018-01-13)
Beatrice Paola Festa, Zhiyong Chen, Marine Berquez, Huguette Debaix, Natsuko Tokonami, Jenny Ann Prange, Glenn van de Hoek, Cremonesi Alessio, Andrea Raimondi, Nathalie Nevo, Rachel H Giles, Olivier Devuyst, Alessandro Luciani
ABSTRACT

The endolysosomal system sustains the reabsorptive activity of specialized epithelial cells. Lysosomal storage diseases such as nephropathic cystinosis cause a major dysfunction of epithelial cells lining the kidney tubule, resulting in massive losses of vital solutes in the urine. The mechanisms linking lysosomal defects and epithelial dysfunction remain unknown, preventing the development of disease-modifying therapies. Here we demonstrate, by combining genetic and pharmacologic approaches, that lysosomal dysfunction in cystinosis results in defective autophagy-mediated clearance of damaged mitochondria. This promotes the generation of oxidative stress that stimulates Gα12/Src-mediated phosphorylation of tight junction ZO-1 and triggers a signaling cascade involving ZO-1-associated Y-box factor ZONAB, which leads to cell proliferation and transport defects. Correction of the primary lysosomal defect, neutralization of mitochondrial oxidative stress, and blockage of tight junction-associated ZONAB signaling rescue the epithelial function. We suggest a link between defective lysosome-autophagy degradation pathways and epithelial dysfunction, providing new therapeutic perspectives for lysosomal storage disorders.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
HEPES solution, 1 M, pH 7.0-7.6, sterile-filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
Paraformaldehyde, powder, 95%
SAFC
MEM Non-essential Amino Acid Solution (100×), without L-glutamine, liquid, sterile-filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
Monoclonal Anti-α-Tubulin antibody produced in mouse, ascites fluid, clone B-5-1-2
Sigma-Aldrich
Monoclonal Anti-β-Actin antibody produced in mouse, clone AC-74, purified immunoglobulin, buffered aqueous solution
Sigma-Aldrich
Dulbecco′s Phosphate Buffered Saline, With calcium chloride and magnesium chloride, 10×, liquid, sterile-filtered, suitable for cell culture
Millipore
ProteinA-Sepharose® 4, Fast Flow from Staphylococcus aureus, aqueous ethanol suspension
Sigma-Aldrich
Anti-Calnexin antibody produced in rabbit, IgG fraction of antiserum, buffered aqueous solution
Sigma-Aldrich
Ethyl 3-aminobenzoate, 97%
Sigma-Aldrich
Anti-ATG7 antibody produced in rabbit, affinity isolated antibody, buffered aqueous solution
Sigma-Aldrich
Anti-YBX3 antibody produced in rabbit, Prestige Antibodies® Powered by Atlas Antibodies, affinity isolated antibody, buffered aqueous glycerol solution
Nunc® Lab-Tek® Chamber Slide system, 8 wells, Permanox® slide, 0.8 cm2/well, sterile, 96/cs