Clioquinol kills astrocyte-derived KT-5 cells by the impairment of the autophagy-lysosome pathway.

Abstract

Clioquinol has been implicated as a causative agent for subacute myelo-optico-neuropathy (SMON) in humans, although the mechanism remains to be elucidated. In this study, we utilized astrocyte-derived cell line, KT-5 cells to explore its potential cytotoxicity on glial cells. KT-5 cells were exposed in vitro to a maximum of 50 μM clioquinol for up to 24 h. 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenylte trazolium bromide (MTT) assay of the cells revealed that clioquinol induced significant cell damage and death. We also found that clioquinol caused accumulation of microtubule-associated protein light chain-3 (LC3)-II and sequestosome-1 (p62) in a dose- and time-dependent manner, suggesting the abnormality of autophagy-lysosome pathway. Consistent with these findings, an exposure of 20 μM clioquinol induced the accumulation of cellular autophagic vacuoles. Moreover, an exposure of 20 μM clioquinol provoked a statistically significant reduction of intracellular lysosomal acid hydrolases activities but no change in lysosomal pH. It also resulted in a significant decline of intracellular ATP levels, enhanced cellular levels of reactive oxygen species, and eventually cell death. This cell death at least did not appear to occur via apoptosis. 10 μM Chloroquine, lysosomal inhibitor, blocked the autophagic degradation and augmented clioquinol-cytotoxicity, whereas rapamycin, an inducer of autophagy, rescued clioquinol-induced cytotoxicity. Thus, our present results strongly suggest clioquinol acts as a potentially cytotoxic agent to glial cells. For future clinical application of clioquinol on the treatment of neurological and cancer disorders, we should take account of this type of cell death mechanism.

• Authors: Yasuaki Mizutani, Toshiki Maeda, Kenichiro Murate, Shinji Ito, Hirohisa Watanabe, Tatsuro Mutoh
• Journal: Archives of toxicology (Arch Toxicol) Vol. 95 Issue 2 Pg. 631-640 (02 2021) ISSN: 1432-0738 [Electronic] Germany
• PMID: 33156368 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• MAP1LC3B protein, human
• Microtubule-Associated Proteins
• Reactive Oxygen Species
• SQSTM1 protein, human
• Sequestosome-1 Protein
• Clioquinol
• Chloroquine
• Adenosine Triphosphate

Topics

• Adenosine Triphosphate (metabolism)
• Apoptosis
• Astrocytes (drug effects, metabolism)
• Autophagy (drug effects)
• Cell Line
• Chloroquine (pharmacology)
• Clioquinol (toxicity)
• Dose-Response Relationship, Drug
• Humans
• Lysosomes (drug effects)
• Microtubule-Associated Proteins (metabolism)
• Neuroglia (drug effects)
• Reactive Oxygen Species (metabolism)
• Sequestosome-1 Protein (metabolism)
• Time Factors