TRAIL-based high throughput screening reveals a link between TRAIL-mediated apoptosis and glutathione reductase, a key component of oxidative stress response

Dmitri Rozanov, Anton Cheltsov, Eduard Sergienko, Stefan Vasile, Vladislav Golubkov, Alexander E. Aleshin, Trevor Levin, Elie Traer, Byron Hann, Julia Freimuth, Nikita Alexeev, Max A. Alekseyev, Sergey P. Budko, Hans Peter Ba¨chinger, Paul Spellman

Research output: Contribution to journalArticle

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Abstract

A high throughput screen for compounds that induce TRAIL-mediated apoptosis identified ML100 as an active chemical probe, which potentiated TRAIL activity in prostate carcinoma PPC-1 and melanoma MDA-MB-435 cells. Follow-up in silico modeling and profiling in cell-based assays allowed us to identify NSC130362, pharmacophore analog of ML100 that induced 65-95% cytotoxicity in cancer cells and did not affect the viability of human primary hepatocytes. In agreement with the activation of the apoptotic pathway, both ML100 and NSC130362 synergistically with TRAIL induced caspase-3/7 activity in MDA-MB-435 cells. Subsequent affinity chromatography and inhibition studies convincingly demonstrated that glutathione reductase (GSR), a key component of the oxidative stress response, is a target of NSC130362. In accordance with the role of GSR in the TRAIL pathway, GSR gene silencing potentiated TRAIL activity in MDA-MB-435 cells but not in human hepatocytes. Inhibition of GSR activity resulted in the induction of oxidative stress, as was evidenced by an increase in intracellular reactive oxygen species (ROS) and peroxidation of mitochondrial membrane after NSC130362 treatment in MDA-MB-435 cells but not in human hepatocytes. The antioxidant reduced glutathione (GSH) fully protected MDA-MB-435 cells from cell lysis induced by NSC130362 and TRAIL, thereby further confirming the interplay between GSR and TRAIL. As a consequence of activation of oxidative stress, combined treatment of different oxidative stress inducers and NSC130362 promoted cell death in a variety of cancer cells but not in hepatocytes in cell-based assays and in in vivo, in a mouse tumor xenograft model.

Original languageEnglish (US)
Article numbere0129566
JournalPLoS One
Volume10
Issue number6
DOIs
StatePublished - Jun 15 2015

Fingerprint

Oxidative stress
Glutathione Reductase
glutathione-disulfide reductase
stress response
Screening
Oxidative Stress
oxidative stress
apoptosis
Throughput
Apoptosis
screening
hepatocytes
Assays
Chemical activation
Cells
cells
Hepatocytes
Affinity chromatography
Cell death
Cytotoxicity

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

TRAIL-based high throughput screening reveals a link between TRAIL-mediated apoptosis and glutathione reductase, a key component of oxidative stress response. / Rozanov, Dmitri; Cheltsov, Anton; Sergienko, Eduard; Vasile, Stefan; Golubkov, Vladislav; Aleshin, Alexander E.; Levin, Trevor; Traer, Elie; Hann, Byron; Freimuth, Julia; Alexeev, Nikita; Alekseyev, Max A.; Budko, Sergey P.; Ba¨chinger, Hans Peter; Spellman, Paul.

In: PLoS One, Vol. 10, No. 6, e0129566, 15.06.2015.

Research output: Contribution to journalArticle

Rozanov, D, Cheltsov, A, Sergienko, E, Vasile, S, Golubkov, V, Aleshin, AE, Levin, T, Traer, E, Hann, B, Freimuth, J, Alexeev, N, Alekseyev, MA, Budko, SP, Ba¨chinger, HP & Spellman, P 2015, 'TRAIL-based high throughput screening reveals a link between TRAIL-mediated apoptosis and glutathione reductase, a key component of oxidative stress response', PLoS One, vol. 10, no. 6, e0129566. https://doi.org/10.1371/journal.pone.0129566
Rozanov, Dmitri ; Cheltsov, Anton ; Sergienko, Eduard ; Vasile, Stefan ; Golubkov, Vladislav ; Aleshin, Alexander E. ; Levin, Trevor ; Traer, Elie ; Hann, Byron ; Freimuth, Julia ; Alexeev, Nikita ; Alekseyev, Max A. ; Budko, Sergey P. ; Ba¨chinger, Hans Peter ; Spellman, Paul. / TRAIL-based high throughput screening reveals a link between TRAIL-mediated apoptosis and glutathione reductase, a key component of oxidative stress response. In: PLoS One. 2015 ; Vol. 10, No. 6.
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abstract = "A high throughput screen for compounds that induce TRAIL-mediated apoptosis identified ML100 as an active chemical probe, which potentiated TRAIL activity in prostate carcinoma PPC-1 and melanoma MDA-MB-435 cells. Follow-up in silico modeling and profiling in cell-based assays allowed us to identify NSC130362, pharmacophore analog of ML100 that induced 65-95{\%} cytotoxicity in cancer cells and did not affect the viability of human primary hepatocytes. In agreement with the activation of the apoptotic pathway, both ML100 and NSC130362 synergistically with TRAIL induced caspase-3/7 activity in MDA-MB-435 cells. Subsequent affinity chromatography and inhibition studies convincingly demonstrated that glutathione reductase (GSR), a key component of the oxidative stress response, is a target of NSC130362. In accordance with the role of GSR in the TRAIL pathway, GSR gene silencing potentiated TRAIL activity in MDA-MB-435 cells but not in human hepatocytes. Inhibition of GSR activity resulted in the induction of oxidative stress, as was evidenced by an increase in intracellular reactive oxygen species (ROS) and peroxidation of mitochondrial membrane after NSC130362 treatment in MDA-MB-435 cells but not in human hepatocytes. The antioxidant reduced glutathione (GSH) fully protected MDA-MB-435 cells from cell lysis induced by NSC130362 and TRAIL, thereby further confirming the interplay between GSR and TRAIL. As a consequence of activation of oxidative stress, combined treatment of different oxidative stress inducers and NSC130362 promoted cell death in a variety of cancer cells but not in hepatocytes in cell-based assays and in in vivo, in a mouse tumor xenograft model.",
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AU - Vasile, Stefan

AU - Golubkov, Vladislav

AU - Aleshin, Alexander E.

AU - Levin, Trevor

AU - Traer, Elie

AU - Hann, Byron

AU - Freimuth, Julia

AU - Alexeev, Nikita

AU - Alekseyev, Max A.

AU - Budko, Sergey P.

AU - Ba¨chinger, Hans Peter

AU - Spellman, Paul

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