Lysosomal adaptation: How cells respond to lysosomotropic compounds

PLoS One. 2017 Mar 16;12(3):e0173771. doi: 10.1371/journal.pone.0173771. eCollection 2017.

Abstract

Lysosomes are acidic organelles essential for degradation and cellular homoeostasis and recently lysosomes have been shown as signaling hub to respond to the intra and extracellular changes (e.g. amino acid availability). Compounds including pharmaceutical drugs that are basic and lipophilic will become sequestered inside lysosomes (lysosomotropic). How cells respond to the lysosomal stress associated with lysosomotropism is not well characterized. Our goal is to assess the lysosomal changes and identify the signaling pathways that involve in the lysosomal changes. Eight chemically diverse lysosomotropic drugs from different therapeutic areas were subjected to the evaluation using the human adult retinal pigmented epithelium cell line, ARPE-19. All lysosomotropic drugs tested triggered lysosomal activation demonstrated by increased lysosotracker red (LTR) and lysosensor green staining, increased cathepsin activity, and increased LAMP2 staining. However, tested lysosomotropic drugs also prompted lysosomal dysfunction exemplified by intracellular and extracellular substrate accumulation including phospholipid, SQSTM1/p62, GAPDH (Glyceraldehyde 3-phosphate dehydrogenase) and opsin. Lysosomal activation observed was likely attributed to lysosomal dysfunction, leading to compensatory responses including nuclear translocation of transcriptional factors TFEB, TFE3 and MITF. The adaptive changes are protective to the cells under lysosomal stress. Mechanistic studies implicate calcium and mTORC1 modulation involvement in the adaptive changes. These results indicate that lysosomotropic compounds could evoke a compensatory lysosomal biogenic response but with the ultimate consequence of lysosomal functional impairment. This work also highlights a pathway of response to lysosomal stress and evidences the role of TFEB, TFE3 and MITF in the stress response.

MeSH terms

  • Adaptation, Physiological*
  • Cell Line
  • Glyceraldehyde-3-Phosphate Dehydrogenases / metabolism
  • Humans
  • Hydrogen-Ion Concentration
  • Lysosomes / drug effects*
  • Lysosomes / enzymology
  • Lysosomes / metabolism
  • Lysosomes / physiology
  • Opsins / metabolism
  • Retinal Pigment Epithelium / cytology
  • Sequestosome-1 Protein / metabolism

Substances

  • Opsins
  • SQSTM1 protein, human
  • Sequestosome-1 Protein
  • Glyceraldehyde-3-Phosphate Dehydrogenases

Grant support

SL, TS, RTA and BAJ are employees of Pfizer Inc. NL was a postdoc in Pfizer Inc. when generating the data. Pfizer provided support in the form of salaries for authors SL, TS, NL, RTA, BAJ but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the author contributions section.