Sphingosine analogue drug FTY720 targets I2PP2A/SET and mediates lung tumour suppression via activation of PP2A-RIPK1-dependent necroptosis

Sahar A. Saddoughi, Salih Gencer, Yuri K. Peterson, Katherine E. Ward, Archana Mukhopadhyay, Joshua Oaks, Jacek Bielawski, Zdzislaw M. Szulc, Raquela J. Thomas, Shanmugam P. Selvam, Can E. Senkal, Elizabeth Garrett-Mayer, Ryan M. De Palma, Dzmitry Fedarovich, Angen Liu, Amyn A. Habib, Robert Stahelin, Danilo Perrotti, Besim Ogretmen

Research output: Contribution to journalArticle

132 Citations (Scopus)

Abstract

Mechanisms that alter protein phosphatase 2A (PP2A)-dependent lung tumour suppression via the I2PP2A/SET oncoprotein are unknown. We show here that the tumour suppressor ceramide binds I2PP2A/SET selectively in the nucleus and including its K209 and Y122 residues as determined by molecular modelling/simulations and site-directed mutagenesis. Because I2PP2A/SET was found overexpressed, whereas ceramide was downregulated in lung tumours, a sphingolipid analogue drug, FTY720, was identified to mimick ceramide for binding and targeting I2PP2A/SET, leading to PP2A reactivation, lung cancer cell death, and tumour suppression in vivo. Accordingly, while molecular targeting of I2PP2A/SET by stable knockdown prevented further tumour suppression by FTY720, reconstitution of WT-I2PP2A/SET expression restored this process. Mechanistically, targeting I2PP2A/SET by FTY720 mediated PP2A/RIPK1-dependent programmed necrosis (necroptosis), but not by apoptosis. The RIPK1 inhibitor necrostatin and knockdown or genetic loss of RIPK1 prevented growth inhibition by FTY720. Expression of WT- or death-domain-deleted (DDD)-RIPK1, but not the kinase-domain-deleted (KDD)-RIPK1, restored FTY720-mediated necroptosis in RIPK1-/- MEFs. Thus, these data suggest that targeting I2PP2A/SET by FTY720 suppresses lung tumour growth, at least in part, via PP2A activation and necroptosis mediated by the kinase domain of RIPK1.

Original languageEnglish
Pages (from-to)105-121
Number of pages17
JournalEMBO Molecular Medicine
Volume5
Issue number1
DOIs
StatePublished - Jan 2013

Fingerprint

Protein Phosphatase 2
Sphingosine
Lung
Ceramides
Pharmaceutical Preparations
Neoplasms
Phosphotransferases
Sphingolipids
Oncogene Proteins
Growth
Site-Directed Mutagenesis
Fingolimod Hydrochloride
Lung Neoplasms
Cell Death
Down-Regulation
Apoptosis

Keywords

  • Ceramide
  • FTY720
  • Sphingolipids
  • Sphingosine
  • Sphingosine kinase 2

ASJC Scopus subject areas

  • Molecular Medicine

Cite this

Sphingosine analogue drug FTY720 targets I2PP2A/SET and mediates lung tumour suppression via activation of PP2A-RIPK1-dependent necroptosis. / Saddoughi, Sahar A.; Gencer, Salih; Peterson, Yuri K.; Ward, Katherine E.; Mukhopadhyay, Archana; Oaks, Joshua; Bielawski, Jacek; Szulc, Zdzislaw M.; Thomas, Raquela J.; Selvam, Shanmugam P.; Senkal, Can E.; Garrett-Mayer, Elizabeth; De Palma, Ryan M.; Fedarovich, Dzmitry; Liu, Angen; Habib, Amyn A.; Stahelin, Robert; Perrotti, Danilo; Ogretmen, Besim.

In: EMBO Molecular Medicine, Vol. 5, No. 1, 01.2013, p. 105-121.

Research output: Contribution to journalArticle

Saddoughi, SA, Gencer, S, Peterson, YK, Ward, KE, Mukhopadhyay, A, Oaks, J, Bielawski, J, Szulc, ZM, Thomas, RJ, Selvam, SP, Senkal, CE, Garrett-Mayer, E, De Palma, RM, Fedarovich, D, Liu, A, Habib, AA, Stahelin, R, Perrotti, D & Ogretmen, B 2013, 'Sphingosine analogue drug FTY720 targets I2PP2A/SET and mediates lung tumour suppression via activation of PP2A-RIPK1-dependent necroptosis', EMBO Molecular Medicine, vol. 5, no. 1, pp. 105-121. https://doi.org/10.1002/emmm.201201283
Saddoughi, Sahar A. ; Gencer, Salih ; Peterson, Yuri K. ; Ward, Katherine E. ; Mukhopadhyay, Archana ; Oaks, Joshua ; Bielawski, Jacek ; Szulc, Zdzislaw M. ; Thomas, Raquela J. ; Selvam, Shanmugam P. ; Senkal, Can E. ; Garrett-Mayer, Elizabeth ; De Palma, Ryan M. ; Fedarovich, Dzmitry ; Liu, Angen ; Habib, Amyn A. ; Stahelin, Robert ; Perrotti, Danilo ; Ogretmen, Besim. / Sphingosine analogue drug FTY720 targets I2PP2A/SET and mediates lung tumour suppression via activation of PP2A-RIPK1-dependent necroptosis. In: EMBO Molecular Medicine. 2013 ; Vol. 5, No. 1. pp. 105-121.
@article{cc3f9d90a8a842d289338218199af1ae,
title = "Sphingosine analogue drug FTY720 targets I2PP2A/SET and mediates lung tumour suppression via activation of PP2A-RIPK1-dependent necroptosis",
abstract = "Mechanisms that alter protein phosphatase 2A (PP2A)-dependent lung tumour suppression via the I2PP2A/SET oncoprotein are unknown. We show here that the tumour suppressor ceramide binds I2PP2A/SET selectively in the nucleus and including its K209 and Y122 residues as determined by molecular modelling/simulations and site-directed mutagenesis. Because I2PP2A/SET was found overexpressed, whereas ceramide was downregulated in lung tumours, a sphingolipid analogue drug, FTY720, was identified to mimick ceramide for binding and targeting I2PP2A/SET, leading to PP2A reactivation, lung cancer cell death, and tumour suppression in vivo. Accordingly, while molecular targeting of I2PP2A/SET by stable knockdown prevented further tumour suppression by FTY720, reconstitution of WT-I2PP2A/SET expression restored this process. Mechanistically, targeting I2PP2A/SET by FTY720 mediated PP2A/RIPK1-dependent programmed necrosis (necroptosis), but not by apoptosis. The RIPK1 inhibitor necrostatin and knockdown or genetic loss of RIPK1 prevented growth inhibition by FTY720. Expression of WT- or death-domain-deleted (DDD)-RIPK1, but not the kinase-domain-deleted (KDD)-RIPK1, restored FTY720-mediated necroptosis in RIPK1-/- MEFs. Thus, these data suggest that targeting I2PP2A/SET by FTY720 suppresses lung tumour growth, at least in part, via PP2A activation and necroptosis mediated by the kinase domain of RIPK1.",
keywords = "Ceramide, FTY720, Sphingolipids, Sphingosine, Sphingosine kinase 2",
author = "Saddoughi, {Sahar A.} and Salih Gencer and Peterson, {Yuri K.} and Ward, {Katherine E.} and Archana Mukhopadhyay and Joshua Oaks and Jacek Bielawski and Szulc, {Zdzislaw M.} and Thomas, {Raquela J.} and Selvam, {Shanmugam P.} and Senkal, {Can E.} and Elizabeth Garrett-Mayer and {De Palma}, {Ryan M.} and Dzmitry Fedarovich and Angen Liu and Habib, {Amyn A.} and Robert Stahelin and Danilo Perrotti and Besim Ogretmen",
year = "2013",
month = "1",
doi = "10.1002/emmm.201201283",
language = "English",
volume = "5",
pages = "105--121",
journal = "EMBO Molecular Medicine",
issn = "1757-4676",
publisher = "Wiley-Blackwell",
number = "1",

}

TY - JOUR

T1 - Sphingosine analogue drug FTY720 targets I2PP2A/SET and mediates lung tumour suppression via activation of PP2A-RIPK1-dependent necroptosis

AU - Saddoughi, Sahar A.

AU - Gencer, Salih

AU - Peterson, Yuri K.

AU - Ward, Katherine E.

AU - Mukhopadhyay, Archana

AU - Oaks, Joshua

AU - Bielawski, Jacek

AU - Szulc, Zdzislaw M.

AU - Thomas, Raquela J.

AU - Selvam, Shanmugam P.

AU - Senkal, Can E.

AU - Garrett-Mayer, Elizabeth

AU - De Palma, Ryan M.

AU - Fedarovich, Dzmitry

AU - Liu, Angen

AU - Habib, Amyn A.

AU - Stahelin, Robert

AU - Perrotti, Danilo

AU - Ogretmen, Besim

PY - 2013/1

Y1 - 2013/1

N2 - Mechanisms that alter protein phosphatase 2A (PP2A)-dependent lung tumour suppression via the I2PP2A/SET oncoprotein are unknown. We show here that the tumour suppressor ceramide binds I2PP2A/SET selectively in the nucleus and including its K209 and Y122 residues as determined by molecular modelling/simulations and site-directed mutagenesis. Because I2PP2A/SET was found overexpressed, whereas ceramide was downregulated in lung tumours, a sphingolipid analogue drug, FTY720, was identified to mimick ceramide for binding and targeting I2PP2A/SET, leading to PP2A reactivation, lung cancer cell death, and tumour suppression in vivo. Accordingly, while molecular targeting of I2PP2A/SET by stable knockdown prevented further tumour suppression by FTY720, reconstitution of WT-I2PP2A/SET expression restored this process. Mechanistically, targeting I2PP2A/SET by FTY720 mediated PP2A/RIPK1-dependent programmed necrosis (necroptosis), but not by apoptosis. The RIPK1 inhibitor necrostatin and knockdown or genetic loss of RIPK1 prevented growth inhibition by FTY720. Expression of WT- or death-domain-deleted (DDD)-RIPK1, but not the kinase-domain-deleted (KDD)-RIPK1, restored FTY720-mediated necroptosis in RIPK1-/- MEFs. Thus, these data suggest that targeting I2PP2A/SET by FTY720 suppresses lung tumour growth, at least in part, via PP2A activation and necroptosis mediated by the kinase domain of RIPK1.

AB - Mechanisms that alter protein phosphatase 2A (PP2A)-dependent lung tumour suppression via the I2PP2A/SET oncoprotein are unknown. We show here that the tumour suppressor ceramide binds I2PP2A/SET selectively in the nucleus and including its K209 and Y122 residues as determined by molecular modelling/simulations and site-directed mutagenesis. Because I2PP2A/SET was found overexpressed, whereas ceramide was downregulated in lung tumours, a sphingolipid analogue drug, FTY720, was identified to mimick ceramide for binding and targeting I2PP2A/SET, leading to PP2A reactivation, lung cancer cell death, and tumour suppression in vivo. Accordingly, while molecular targeting of I2PP2A/SET by stable knockdown prevented further tumour suppression by FTY720, reconstitution of WT-I2PP2A/SET expression restored this process. Mechanistically, targeting I2PP2A/SET by FTY720 mediated PP2A/RIPK1-dependent programmed necrosis (necroptosis), but not by apoptosis. The RIPK1 inhibitor necrostatin and knockdown or genetic loss of RIPK1 prevented growth inhibition by FTY720. Expression of WT- or death-domain-deleted (DDD)-RIPK1, but not the kinase-domain-deleted (KDD)-RIPK1, restored FTY720-mediated necroptosis in RIPK1-/- MEFs. Thus, these data suggest that targeting I2PP2A/SET by FTY720 suppresses lung tumour growth, at least in part, via PP2A activation and necroptosis mediated by the kinase domain of RIPK1.

KW - Ceramide

KW - FTY720

KW - Sphingolipids

KW - Sphingosine

KW - Sphingosine kinase 2

UR - http://www.scopus.com/inward/record.url?scp=84871968824&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84871968824&partnerID=8YFLogxK

U2 - 10.1002/emmm.201201283

DO - 10.1002/emmm.201201283

M3 - Article

VL - 5

SP - 105

EP - 121

JO - EMBO Molecular Medicine

JF - EMBO Molecular Medicine

SN - 1757-4676

IS - 1

ER -