Pharmacological activation of SIRT6 triggers lethal autophagy in human cancer cells

Sara Iachettini, Daniela Trisciuoglio, Dante Rotili, Alessia Lucidi, Erica Salvati, Pasquale Zizza, Luca Di Leo, Donatella Del Bufalo, Maria Rosa Ciriolo, Carlo Leonetti, Clemens Steegborn, Antonello Mai, Angela Rizzo, Annamaria Biroccio

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Sirtuin 6 (SIRT6) is a member of the NAD+-dependent class III deacetylase sirtuin family, which plays a key role in cancer by controlling transcription, genome stability, telomere integrity, DNA repair, and autophagy. Here we analyzed the molecular and biological effects of UBCS039, the first synthetic SIRT6 activator. Our data demonstrated that UBCS039 induced a time-dependent activation of autophagy in several human tumor cell lines, as evaluated by increased content of the lipidated form of LC3B by western blot and of autophagosomal puncta by microscopy analysis of GFP-LC3. UBCS039-mediated activation of autophagy was strictly dependent on SIRT6 deacetylating activity since the catalytic mutant H133Y failed to activate autophagy. At the molecular level, SIRT6-mediated autophagy was triggered by an increase of ROS levels, which, in turn, resulted in the activation of the AMPK-ULK1-mTOR signaling pathway. Interestingly, antioxidants were able to completely counteract UBCS039-induced autophagy, suggesting that ROS burst had a key role in upstream events leading to autophagy commitment. Finally, sustained activation of SIRT6 resulted in autophagy-related cell death, a process that was markedly attenuated using either a pan caspases inhibitor (zVAD-fmk) or an autophagy inhibitor (CQ). Overall, our results identified UBCS039 as an efficient SIRT6 activator, thereby providing a proof of principle that modulation of the enzyme can influence therapeutic strategy by enhancing autophagy-dependent cell death.

Original languageEnglish
Article number996
JournalCell Death and Disease
Volume9
Issue number10
DOIs
Publication statusPublished - Oct 1 2018

Fingerprint

Autophagy
Pharmacology
Neoplasms
Cell Death
AMP-Activated Protein Kinases
Caspase Inhibitors
Genomic Instability
Telomere
Tumor Cell Line
DNA Repair
NAD
Microscopy
Antioxidants
Western Blotting

ASJC Scopus subject areas

  • Immunology
  • Cellular and Molecular Neuroscience
  • Cell Biology
  • Cancer Research

Cite this

Pharmacological activation of SIRT6 triggers lethal autophagy in human cancer cells. / Iachettini, Sara; Trisciuoglio, Daniela; Rotili, Dante; Lucidi, Alessia; Salvati, Erica; Zizza, Pasquale; Di Leo, Luca; Del Bufalo, Donatella; Ciriolo, Maria Rosa; Leonetti, Carlo; Steegborn, Clemens; Mai, Antonello; Rizzo, Angela; Biroccio, Annamaria.

In: Cell Death and Disease, Vol. 9, No. 10, 996, 01.10.2018.

Research output: Contribution to journalArticle

@article{6a821056fefe40c1a711406720aa65df,
title = "Pharmacological activation of SIRT6 triggers lethal autophagy in human cancer cells",
abstract = "Sirtuin 6 (SIRT6) is a member of the NAD+-dependent class III deacetylase sirtuin family, which plays a key role in cancer by controlling transcription, genome stability, telomere integrity, DNA repair, and autophagy. Here we analyzed the molecular and biological effects of UBCS039, the first synthetic SIRT6 activator. Our data demonstrated that UBCS039 induced a time-dependent activation of autophagy in several human tumor cell lines, as evaluated by increased content of the lipidated form of LC3B by western blot and of autophagosomal puncta by microscopy analysis of GFP-LC3. UBCS039-mediated activation of autophagy was strictly dependent on SIRT6 deacetylating activity since the catalytic mutant H133Y failed to activate autophagy. At the molecular level, SIRT6-mediated autophagy was triggered by an increase of ROS levels, which, in turn, resulted in the activation of the AMPK-ULK1-mTOR signaling pathway. Interestingly, antioxidants were able to completely counteract UBCS039-induced autophagy, suggesting that ROS burst had a key role in upstream events leading to autophagy commitment. Finally, sustained activation of SIRT6 resulted in autophagy-related cell death, a process that was markedly attenuated using either a pan caspases inhibitor (zVAD-fmk) or an autophagy inhibitor (CQ). Overall, our results identified UBCS039 as an efficient SIRT6 activator, thereby providing a proof of principle that modulation of the enzyme can influence therapeutic strategy by enhancing autophagy-dependent cell death.",
author = "Sara Iachettini and Daniela Trisciuoglio and Dante Rotili and Alessia Lucidi and Erica Salvati and Pasquale Zizza and {Di Leo}, Luca and {Del Bufalo}, Donatella and Ciriolo, {Maria Rosa} and Carlo Leonetti and Clemens Steegborn and Antonello Mai and Angela Rizzo and Annamaria Biroccio",
year = "2018",
month = "10",
day = "1",
doi = "10.1038/s41419-018-1065-0",
language = "English",
volume = "9",
journal = "Cell Death and Disease",
issn = "2041-4889",
publisher = "Nature Publishing Group",
number = "10",

}

TY - JOUR

T1 - Pharmacological activation of SIRT6 triggers lethal autophagy in human cancer cells

AU - Iachettini, Sara

AU - Trisciuoglio, Daniela

AU - Rotili, Dante

AU - Lucidi, Alessia

AU - Salvati, Erica

AU - Zizza, Pasquale

AU - Di Leo, Luca

AU - Del Bufalo, Donatella

AU - Ciriolo, Maria Rosa

AU - Leonetti, Carlo

AU - Steegborn, Clemens

AU - Mai, Antonello

AU - Rizzo, Angela

AU - Biroccio, Annamaria

PY - 2018/10/1

Y1 - 2018/10/1

N2 - Sirtuin 6 (SIRT6) is a member of the NAD+-dependent class III deacetylase sirtuin family, which plays a key role in cancer by controlling transcription, genome stability, telomere integrity, DNA repair, and autophagy. Here we analyzed the molecular and biological effects of UBCS039, the first synthetic SIRT6 activator. Our data demonstrated that UBCS039 induced a time-dependent activation of autophagy in several human tumor cell lines, as evaluated by increased content of the lipidated form of LC3B by western blot and of autophagosomal puncta by microscopy analysis of GFP-LC3. UBCS039-mediated activation of autophagy was strictly dependent on SIRT6 deacetylating activity since the catalytic mutant H133Y failed to activate autophagy. At the molecular level, SIRT6-mediated autophagy was triggered by an increase of ROS levels, which, in turn, resulted in the activation of the AMPK-ULK1-mTOR signaling pathway. Interestingly, antioxidants were able to completely counteract UBCS039-induced autophagy, suggesting that ROS burst had a key role in upstream events leading to autophagy commitment. Finally, sustained activation of SIRT6 resulted in autophagy-related cell death, a process that was markedly attenuated using either a pan caspases inhibitor (zVAD-fmk) or an autophagy inhibitor (CQ). Overall, our results identified UBCS039 as an efficient SIRT6 activator, thereby providing a proof of principle that modulation of the enzyme can influence therapeutic strategy by enhancing autophagy-dependent cell death.

AB - Sirtuin 6 (SIRT6) is a member of the NAD+-dependent class III deacetylase sirtuin family, which plays a key role in cancer by controlling transcription, genome stability, telomere integrity, DNA repair, and autophagy. Here we analyzed the molecular and biological effects of UBCS039, the first synthetic SIRT6 activator. Our data demonstrated that UBCS039 induced a time-dependent activation of autophagy in several human tumor cell lines, as evaluated by increased content of the lipidated form of LC3B by western blot and of autophagosomal puncta by microscopy analysis of GFP-LC3. UBCS039-mediated activation of autophagy was strictly dependent on SIRT6 deacetylating activity since the catalytic mutant H133Y failed to activate autophagy. At the molecular level, SIRT6-mediated autophagy was triggered by an increase of ROS levels, which, in turn, resulted in the activation of the AMPK-ULK1-mTOR signaling pathway. Interestingly, antioxidants were able to completely counteract UBCS039-induced autophagy, suggesting that ROS burst had a key role in upstream events leading to autophagy commitment. Finally, sustained activation of SIRT6 resulted in autophagy-related cell death, a process that was markedly attenuated using either a pan caspases inhibitor (zVAD-fmk) or an autophagy inhibitor (CQ). Overall, our results identified UBCS039 as an efficient SIRT6 activator, thereby providing a proof of principle that modulation of the enzyme can influence therapeutic strategy by enhancing autophagy-dependent cell death.

UR - http://www.scopus.com/inward/record.url?scp=85053859793&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85053859793&partnerID=8YFLogxK

U2 - 10.1038/s41419-018-1065-0

DO - 10.1038/s41419-018-1065-0

M3 - Article

C2 - 30250025

AN - SCOPUS:85053859793

VL - 9

JO - Cell Death and Disease

JF - Cell Death and Disease

SN - 2041-4889

IS - 10

M1 - 996

ER -