Molecular characterization of PS-341 (bortezomib) resistance: Implications for overcoming resistance using lysophosphatidic acid acyltransferase (LPAAT)-β inhibitors

Teru Hideshima, Dharminder Chauhan, Kenji Ishitsuka, Hiroshi Yasui, Noopur Raje, Shaji K Kumar, Klaus Podar, Constantine Mitsiades, Hiromasa Hideshima, Lynn Bonham, Nikhil C. Munshi, Paul G. Richardson, Jack W. Singer, Kenneth C. Anderson

Research output: Contribution to journalArticle

43 Citations (Scopus)

Abstract

PS-341 (bortezomib, Velcade™) is a promising novel agent for treatment of advanced multiple myeloma (MM); however, 65% of patients with relapsed refractory disease in a phase II study do not respond to PS-341. We have previously shown that lysophosphatidic acid acyltransferase (LPAAT)-β inhibitor CT-32615 triggers caspase-dependent apoptosis, and can overcome resistance to conventional therapeutics (i.e., dexamethasone, doxorubicin, melphalan) in MM cells. In this study, we therefore determined whether CT-32615 could also overcome resistance to PS-341. We first characterized molecular mechanisms of resistance to PS-341 in DHL-4 cells. DHL-4 cells express low levels of caspase-3 and caspase-8; furthermore, no cleavage in caspase-8, caspase-9, caspase-3, poly ADP-ribose polymerase (PARP), or DNA fragmentation factor 45 was triggered by PS-341 treatment. We have previously shown that PS-341 treatment triggers phosphorylation of c-Jun NH2-terminal kinase (JNK), which subsequently induces caspase-dependent apoptosis; conversely, JNK inhibition blocks PS-341-induced apoptosis. We here show that phosphorylation of SEK-1, JNK, and c-Jun are not induced by PS-341 treatment, suggesting that PS-341 does not trigger a stress response in DHL-4 cells. Importantly, CT-32615 inhibits growth of DHL-4 cells in a time- and dose-dependent fashion: a transient G2/M cell cycle arrest induced by CT-32615 is mediated via downregulation of cdc25c and cdc2. CT-32615 triggered swelling and lysis of DHL-4 cells, without caspase/PARP cleavage or TUNEL-positivity, suggesting a necrotic response. Our studies therefore demonstrate that LPAAT-β inhibitor CT-32615 triggers necrosis, even in PS-341-resistant DHL-4 cells, providing the framework for its evaluation to overcome clinical PS-341 resistance and improve patient outcome.

Original languageEnglish (US)
Pages (from-to)3121-3129
Number of pages9
JournalOncogene
Volume24
Issue number19
DOIs
StatePublished - Apr 28 2005
Externally publishedYes

Fingerprint

JNK Mitogen-Activated Protein Kinases
Caspases
Poly(ADP-ribose) Polymerases
Caspase 8
Apoptosis
Multiple Myeloma
Caspase 3
Bortezomib
2-acylglycerophosphate acyltransferase
Phosphorylation
G2 Phase Cell Cycle Checkpoints
Therapeutics
Melphalan
Caspase 9
In Situ Nick-End Labeling
Doxorubicin
Dexamethasone
Necrosis
Down-Regulation
Growth

Keywords

  • Bortezomib
  • Caspase
  • Cell cycle
  • Lysophosphatidic acid acyltransferase-β inhibitor
  • Necrosis
  • PS-341

ASJC Scopus subject areas

  • Molecular Biology
  • Cancer Research
  • Genetics

Cite this

Molecular characterization of PS-341 (bortezomib) resistance : Implications for overcoming resistance using lysophosphatidic acid acyltransferase (LPAAT)-β inhibitors. / Hideshima, Teru; Chauhan, Dharminder; Ishitsuka, Kenji; Yasui, Hiroshi; Raje, Noopur; Kumar, Shaji K; Podar, Klaus; Mitsiades, Constantine; Hideshima, Hiromasa; Bonham, Lynn; Munshi, Nikhil C.; Richardson, Paul G.; Singer, Jack W.; Anderson, Kenneth C.

In: Oncogene, Vol. 24, No. 19, 28.04.2005, p. 3121-3129.

Research output: Contribution to journalArticle

Hideshima, T, Chauhan, D, Ishitsuka, K, Yasui, H, Raje, N, Kumar, SK, Podar, K, Mitsiades, C, Hideshima, H, Bonham, L, Munshi, NC, Richardson, PG, Singer, JW & Anderson, KC 2005, 'Molecular characterization of PS-341 (bortezomib) resistance: Implications for overcoming resistance using lysophosphatidic acid acyltransferase (LPAAT)-β inhibitors', Oncogene, vol. 24, no. 19, pp. 3121-3129. https://doi.org/10.1038/sj.onc.1208522
Hideshima, Teru ; Chauhan, Dharminder ; Ishitsuka, Kenji ; Yasui, Hiroshi ; Raje, Noopur ; Kumar, Shaji K ; Podar, Klaus ; Mitsiades, Constantine ; Hideshima, Hiromasa ; Bonham, Lynn ; Munshi, Nikhil C. ; Richardson, Paul G. ; Singer, Jack W. ; Anderson, Kenneth C. / Molecular characterization of PS-341 (bortezomib) resistance : Implications for overcoming resistance using lysophosphatidic acid acyltransferase (LPAAT)-β inhibitors. In: Oncogene. 2005 ; Vol. 24, No. 19. pp. 3121-3129.
@article{c2f0118e142d4aa8a2794e4b204bcba6,
title = "Molecular characterization of PS-341 (bortezomib) resistance: Implications for overcoming resistance using lysophosphatidic acid acyltransferase (LPAAT)-β inhibitors",
abstract = "PS-341 (bortezomib, Velcade™) is a promising novel agent for treatment of advanced multiple myeloma (MM); however, 65{\%} of patients with relapsed refractory disease in a phase II study do not respond to PS-341. We have previously shown that lysophosphatidic acid acyltransferase (LPAAT)-β inhibitor CT-32615 triggers caspase-dependent apoptosis, and can overcome resistance to conventional therapeutics (i.e., dexamethasone, doxorubicin, melphalan) in MM cells. In this study, we therefore determined whether CT-32615 could also overcome resistance to PS-341. We first characterized molecular mechanisms of resistance to PS-341 in DHL-4 cells. DHL-4 cells express low levels of caspase-3 and caspase-8; furthermore, no cleavage in caspase-8, caspase-9, caspase-3, poly ADP-ribose polymerase (PARP), or DNA fragmentation factor 45 was triggered by PS-341 treatment. We have previously shown that PS-341 treatment triggers phosphorylation of c-Jun NH2-terminal kinase (JNK), which subsequently induces caspase-dependent apoptosis; conversely, JNK inhibition blocks PS-341-induced apoptosis. We here show that phosphorylation of SEK-1, JNK, and c-Jun are not induced by PS-341 treatment, suggesting that PS-341 does not trigger a stress response in DHL-4 cells. Importantly, CT-32615 inhibits growth of DHL-4 cells in a time- and dose-dependent fashion: a transient G2/M cell cycle arrest induced by CT-32615 is mediated via downregulation of cdc25c and cdc2. CT-32615 triggered swelling and lysis of DHL-4 cells, without caspase/PARP cleavage or TUNEL-positivity, suggesting a necrotic response. Our studies therefore demonstrate that LPAAT-β inhibitor CT-32615 triggers necrosis, even in PS-341-resistant DHL-4 cells, providing the framework for its evaluation to overcome clinical PS-341 resistance and improve patient outcome.",
keywords = "Bortezomib, Caspase, Cell cycle, Lysophosphatidic acid acyltransferase-β inhibitor, Necrosis, PS-341",
author = "Teru Hideshima and Dharminder Chauhan and Kenji Ishitsuka and Hiroshi Yasui and Noopur Raje and Kumar, {Shaji K} and Klaus Podar and Constantine Mitsiades and Hiromasa Hideshima and Lynn Bonham and Munshi, {Nikhil C.} and Richardson, {Paul G.} and Singer, {Jack W.} and Anderson, {Kenneth C.}",
year = "2005",
month = "4",
day = "28",
doi = "10.1038/sj.onc.1208522",
language = "English (US)",
volume = "24",
pages = "3121--3129",
journal = "Oncogene",
issn = "0950-9232",
publisher = "Nature Publishing Group",
number = "19",

}

TY - JOUR

T1 - Molecular characterization of PS-341 (bortezomib) resistance

T2 - Implications for overcoming resistance using lysophosphatidic acid acyltransferase (LPAAT)-β inhibitors

AU - Hideshima, Teru

AU - Chauhan, Dharminder

AU - Ishitsuka, Kenji

AU - Yasui, Hiroshi

AU - Raje, Noopur

AU - Kumar, Shaji K

AU - Podar, Klaus

AU - Mitsiades, Constantine

AU - Hideshima, Hiromasa

AU - Bonham, Lynn

AU - Munshi, Nikhil C.

AU - Richardson, Paul G.

AU - Singer, Jack W.

AU - Anderson, Kenneth C.

PY - 2005/4/28

Y1 - 2005/4/28

N2 - PS-341 (bortezomib, Velcade™) is a promising novel agent for treatment of advanced multiple myeloma (MM); however, 65% of patients with relapsed refractory disease in a phase II study do not respond to PS-341. We have previously shown that lysophosphatidic acid acyltransferase (LPAAT)-β inhibitor CT-32615 triggers caspase-dependent apoptosis, and can overcome resistance to conventional therapeutics (i.e., dexamethasone, doxorubicin, melphalan) in MM cells. In this study, we therefore determined whether CT-32615 could also overcome resistance to PS-341. We first characterized molecular mechanisms of resistance to PS-341 in DHL-4 cells. DHL-4 cells express low levels of caspase-3 and caspase-8; furthermore, no cleavage in caspase-8, caspase-9, caspase-3, poly ADP-ribose polymerase (PARP), or DNA fragmentation factor 45 was triggered by PS-341 treatment. We have previously shown that PS-341 treatment triggers phosphorylation of c-Jun NH2-terminal kinase (JNK), which subsequently induces caspase-dependent apoptosis; conversely, JNK inhibition blocks PS-341-induced apoptosis. We here show that phosphorylation of SEK-1, JNK, and c-Jun are not induced by PS-341 treatment, suggesting that PS-341 does not trigger a stress response in DHL-4 cells. Importantly, CT-32615 inhibits growth of DHL-4 cells in a time- and dose-dependent fashion: a transient G2/M cell cycle arrest induced by CT-32615 is mediated via downregulation of cdc25c and cdc2. CT-32615 triggered swelling and lysis of DHL-4 cells, without caspase/PARP cleavage or TUNEL-positivity, suggesting a necrotic response. Our studies therefore demonstrate that LPAAT-β inhibitor CT-32615 triggers necrosis, even in PS-341-resistant DHL-4 cells, providing the framework for its evaluation to overcome clinical PS-341 resistance and improve patient outcome.

AB - PS-341 (bortezomib, Velcade™) is a promising novel agent for treatment of advanced multiple myeloma (MM); however, 65% of patients with relapsed refractory disease in a phase II study do not respond to PS-341. We have previously shown that lysophosphatidic acid acyltransferase (LPAAT)-β inhibitor CT-32615 triggers caspase-dependent apoptosis, and can overcome resistance to conventional therapeutics (i.e., dexamethasone, doxorubicin, melphalan) in MM cells. In this study, we therefore determined whether CT-32615 could also overcome resistance to PS-341. We first characterized molecular mechanisms of resistance to PS-341 in DHL-4 cells. DHL-4 cells express low levels of caspase-3 and caspase-8; furthermore, no cleavage in caspase-8, caspase-9, caspase-3, poly ADP-ribose polymerase (PARP), or DNA fragmentation factor 45 was triggered by PS-341 treatment. We have previously shown that PS-341 treatment triggers phosphorylation of c-Jun NH2-terminal kinase (JNK), which subsequently induces caspase-dependent apoptosis; conversely, JNK inhibition blocks PS-341-induced apoptosis. We here show that phosphorylation of SEK-1, JNK, and c-Jun are not induced by PS-341 treatment, suggesting that PS-341 does not trigger a stress response in DHL-4 cells. Importantly, CT-32615 inhibits growth of DHL-4 cells in a time- and dose-dependent fashion: a transient G2/M cell cycle arrest induced by CT-32615 is mediated via downregulation of cdc25c and cdc2. CT-32615 triggered swelling and lysis of DHL-4 cells, without caspase/PARP cleavage or TUNEL-positivity, suggesting a necrotic response. Our studies therefore demonstrate that LPAAT-β inhibitor CT-32615 triggers necrosis, even in PS-341-resistant DHL-4 cells, providing the framework for its evaluation to overcome clinical PS-341 resistance and improve patient outcome.

KW - Bortezomib

KW - Caspase

KW - Cell cycle

KW - Lysophosphatidic acid acyltransferase-β inhibitor

KW - Necrosis

KW - PS-341

UR - http://www.scopus.com/inward/record.url?scp=21044446935&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=21044446935&partnerID=8YFLogxK

U2 - 10.1038/sj.onc.1208522

DO - 10.1038/sj.onc.1208522

M3 - Article

C2 - 15735676

AN - SCOPUS:21044446935

VL - 24

SP - 3121

EP - 3129

JO - Oncogene

JF - Oncogene

SN - 0950-9232

IS - 19

ER -