Verapamil treatment induces cytoprotective autophagy by modulating cellular metabolism.
Verapamil, an L-type calcium channel blocker, has been used successfully to treat cardiovascular diseases. Interestingly, we have recently shown that treatment of cancer cells with verapamil causes an effect on autophagy. As autophagy is known to modulate chemotherapy responses, this prompted us to explore the impact of verapamil on autophagy and cell viability in greater detail. We report here that verapamil causes an induction of autophagic flux in a number or tumor cells and immortalized normal cells. Moreover, we found that inhibition of autophagy in COLO 205 cells, via treatment with the chloroquine (CQ) or by CRISPR/Cas9-mediated disruption of the autophagy genes Atg7 and Atg5, causes an upregulation of apoptotic markers in response to verapamil. In search of a mechanism for this effect and because autophagy can often mitigate metabolic stress, we examined the impact of verapamil on cellular metabolism. This revealed that in normal prostate cells, verapamil diminishes glucose and glycolytic intermediate levels leading to adenosine 5'-triphosphate (ATP) depletion. In contrast, in COLO 205 cells it enhances aerobic glycolysis and maintains ATP. Importantly, we found that the autophagic response in these cells is related to the activity of l-lactate dehydrogenase A (LDHA, EC 1.1.1.27), as inhibition of LDHA reduces both basal and verapamil-induced autophagy and consequently decreases cell viability. In summary, these findings not only identify a novel mechanism of cytoprotective autophagy induction but they also highlight the potential of using verapamil together with inhibitors of autophagy for the treatment of malignant disease. ENZYMES: l-lactate dehydrogenase (LDHA, EC 1.1.1.27).
Mossakowski Medical Research Centre, Polish Academy of Science, Electron Microscopy Platform, Warsaw, Poland.
Elżbieta
E
Kania
Mossakowski Medical Research Centre, Polish Academy of Science, Electron Microscopy Platform, Warsaw, Poland.|Independent Laboratory of Genetics and Molecular Biology, Kaczkowski Military Institute of Hygiene and Epidemiology, Warsaw, Poland.
Beata
B
Pająk
Cancer Research UK Beatson Institute, Glasgow, UK.
Jim
J
O'Prey
Cancer Research UK Beatson Institute, Glasgow, UK.
Pablo
P
Sierra Gonzalez
Department of Neuroendocrinology, Centre of Postgraduate Medical Education, Warsaw, Poland.
Anna
A
Litwiniuk
Warsaw University of Life Sciences - SGGW, Poland.
Kaja
K
Urbańska
Cancer Research UK Beatson Institute, Glasgow, UK.
Kevin M
KM
Ryan
Mossakowski Medical Research Centre, Polish Academy of Science, Electron Microscopy Platform, Warsaw, Poland.|Warsaw University of Life Sciences - SGGW, Poland.
Arkadiusz
A
Orzechowski
D000962:Antimalarials / Q000494:pharmacology; D000970:Antineoplastic Agents / Q000009:adverse effects / Q000494:pharmacology*; D000071182:Autophagosomes / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D001343:Autophagy / Q000187:drug effects*; D000071187:Autophagy-Related Protein 5 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D000071193:Autophagy-Related Protein 7 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism; D064113:CRISPR-Cas Systems; D002121:Calcium Channel Blockers / Q000009:adverse effects / Q000494:pharmacology*; D002461:Cell Line, Transformed; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002738:Chloroquine / Q000494:pharmacology; D019610:Cytoprotection / Q000187:drug effects*; D004734:Energy Metabolism / Q000187:drug effects; D006019:Glycolysis / Q000187:drug effects*; D006801:Humans; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000378:metabolism; D007770:L-Lactate Dehydrogenase / Q000037:antagonists & inhibitors / Q000378:metabolism; D000080864:Lactate Dehydrogenase 5; D046529:Microscopy, Electron, Transmission; D009369:Neoplasms / Q000188:drug therapy* / Q000378:metabolism / Q000648:ultrastructure; D014700:Verapamil / Q000009:adverse effects / Q000494:pharmacology*
C502778:ATG5 protein, human; D000962:Antimalarials; D000970:Antineoplastic Agents; D000071187:Autophagy-Related Protein 5; D015415:Biomarkers; D002121:Calcium Channel Blockers; D007527:Isoenzymes; D002738:Chloroquine; D014700:Verapamil; D007770:L-Lactate Dehydrogenase; D000080864:Lactate Dehydrogenase 5; C509349:ATG7 protein, human; D000071193:Autophagy-Related Protein 7
autophagy; cancer; cell death; lactate; verapamil