Inhibition of histone deacetylation potentiates the evolution of acquired temozolomide resistance linked to MGMT upregulation in glioblastoma xenografts

Gaspar J. Kitange, Ann C. Mladek, Brett L. Carlson, Mark A. Schroeder, Jenny L. Pokorny, Ling Cen, Paul A. Decker, Wenting Wu, Gwen A. Lomberk, Shiv K. Gupta, Raul A. Urrutia, Jann N Sarkaria

Research output: Contribution to journalArticle

70 Citations (Scopus)

Abstract

Purpose: The therapeutic benefit of temozolomide in glioblastoma multiforme (GBM) is limited by resistance. The goal of this study was to elucidate mechanisms of temozolomide resistance in GBM. Experimental Design: We developed an in vivoGBMmodel of temozolomide resistance and used paired parental and temozolomide-resistant tumors to define the mechanisms underlying the development of resistance and the influence of histone deacetylation (HDAC) inhibition. Results: Analysis of paired parental and resistant lines showed upregulation of O6-methylguanine-DNA methyltransferase (MGMT) expression in 3 of the 5 resistant xenografts. While no significant change was detected in MGMT promoter methylation between parental and derivative-resistant samples, chromatin immunoprecipitation showed an association between MGMT upregulation and elevated acetylation of lysine 9 of histone H3 (H3K9-ac) and decreased dimethylation (H3K9-me2) in GBM12 and GBM14. In contrast, temozolomide resistance development in GBM22 was not linked to MGMT expression, and both parental and resistant lines had low H3K9-ac and high H3K9-me2 within the MGMT promoter. In the GBM12TMZ-resistant line, MGMT reexpression was accompanied by increased recruitment of SP1, C-JUN, NF-kB, and p300 within the MGMT promoter. Interestingly, combined treatment of GBM12 flank xenografts with temozolomide and the HDAC inhibitor suberoylanilide hydroxamic acid (SAHA) favored the evolution of temozolomide resistance by MGMT overexpression as compared with treatment with temozolomide alone. Conclusion: This study shows, for the first time, a unique mechanism of temozolomide resistance development driven by chromatin-mediated MGMT upregulation and highlights the potential for epigenetically directed therapies to influence the mechanisms of resistance development in GBM.

Original languageEnglish (US)
Pages (from-to)4070-4079
Number of pages10
JournalClinical Cancer Research
Volume18
Issue number15
DOIs
StatePublished - Aug 1 2012

Fingerprint

temozolomide
Methyltransferases
Glioblastoma
Heterografts
Histones
Up-Regulation
DNA
NF-kappa B
Chromatin Immunoprecipitation
Acetylation

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Inhibition of histone deacetylation potentiates the evolution of acquired temozolomide resistance linked to MGMT upregulation in glioblastoma xenografts. / Kitange, Gaspar J.; Mladek, Ann C.; Carlson, Brett L.; Schroeder, Mark A.; Pokorny, Jenny L.; Cen, Ling; Decker, Paul A.; Wu, Wenting; Lomberk, Gwen A.; Gupta, Shiv K.; Urrutia, Raul A.; Sarkaria, Jann N.

In: Clinical Cancer Research, Vol. 18, No. 15, 01.08.2012, p. 4070-4079.

Research output: Contribution to journalArticle

Kitange, GJ, Mladek, AC, Carlson, BL, Schroeder, MA, Pokorny, JL, Cen, L, Decker, PA, Wu, W, Lomberk, GA, Gupta, SK, Urrutia, RA & Sarkaria, JN 2012, 'Inhibition of histone deacetylation potentiates the evolution of acquired temozolomide resistance linked to MGMT upregulation in glioblastoma xenografts', Clinical Cancer Research, vol. 18, no. 15, pp. 4070-4079. https://doi.org/10.1158/1078-0432.CCR-12-0560
Kitange, Gaspar J. ; Mladek, Ann C. ; Carlson, Brett L. ; Schroeder, Mark A. ; Pokorny, Jenny L. ; Cen, Ling ; Decker, Paul A. ; Wu, Wenting ; Lomberk, Gwen A. ; Gupta, Shiv K. ; Urrutia, Raul A. ; Sarkaria, Jann N. / Inhibition of histone deacetylation potentiates the evolution of acquired temozolomide resistance linked to MGMT upregulation in glioblastoma xenografts. In: Clinical Cancer Research. 2012 ; Vol. 18, No. 15. pp. 4070-4079.
@article{3b98780ff2f9477ab77ec7288846f595,
title = "Inhibition of histone deacetylation potentiates the evolution of acquired temozolomide resistance linked to MGMT upregulation in glioblastoma xenografts",
abstract = "Purpose: The therapeutic benefit of temozolomide in glioblastoma multiforme (GBM) is limited by resistance. The goal of this study was to elucidate mechanisms of temozolomide resistance in GBM. Experimental Design: We developed an in vivoGBMmodel of temozolomide resistance and used paired parental and temozolomide-resistant tumors to define the mechanisms underlying the development of resistance and the influence of histone deacetylation (HDAC) inhibition. Results: Analysis of paired parental and resistant lines showed upregulation of O6-methylguanine-DNA methyltransferase (MGMT) expression in 3 of the 5 resistant xenografts. While no significant change was detected in MGMT promoter methylation between parental and derivative-resistant samples, chromatin immunoprecipitation showed an association between MGMT upregulation and elevated acetylation of lysine 9 of histone H3 (H3K9-ac) and decreased dimethylation (H3K9-me2) in GBM12 and GBM14. In contrast, temozolomide resistance development in GBM22 was not linked to MGMT expression, and both parental and resistant lines had low H3K9-ac and high H3K9-me2 within the MGMT promoter. In the GBM12TMZ-resistant line, MGMT reexpression was accompanied by increased recruitment of SP1, C-JUN, NF-kB, and p300 within the MGMT promoter. Interestingly, combined treatment of GBM12 flank xenografts with temozolomide and the HDAC inhibitor suberoylanilide hydroxamic acid (SAHA) favored the evolution of temozolomide resistance by MGMT overexpression as compared with treatment with temozolomide alone. Conclusion: This study shows, for the first time, a unique mechanism of temozolomide resistance development driven by chromatin-mediated MGMT upregulation and highlights the potential for epigenetically directed therapies to influence the mechanisms of resistance development in GBM.",
author = "Kitange, {Gaspar J.} and Mladek, {Ann C.} and Carlson, {Brett L.} and Schroeder, {Mark A.} and Pokorny, {Jenny L.} and Ling Cen and Decker, {Paul A.} and Wenting Wu and Lomberk, {Gwen A.} and Gupta, {Shiv K.} and Urrutia, {Raul A.} and Sarkaria, {Jann N}",
year = "2012",
month = "8",
day = "1",
doi = "10.1158/1078-0432.CCR-12-0560",
language = "English (US)",
volume = "18",
pages = "4070--4079",
journal = "Clinical Cancer Research",
issn = "1078-0432",
publisher = "American Association for Cancer Research Inc.",
number = "15",

}

TY - JOUR

T1 - Inhibition of histone deacetylation potentiates the evolution of acquired temozolomide resistance linked to MGMT upregulation in glioblastoma xenografts

AU - Kitange, Gaspar J.

AU - Mladek, Ann C.

AU - Carlson, Brett L.

AU - Schroeder, Mark A.

AU - Pokorny, Jenny L.

AU - Cen, Ling

AU - Decker, Paul A.

AU - Wu, Wenting

AU - Lomberk, Gwen A.

AU - Gupta, Shiv K.

AU - Urrutia, Raul A.

AU - Sarkaria, Jann N

PY - 2012/8/1

Y1 - 2012/8/1

N2 - Purpose: The therapeutic benefit of temozolomide in glioblastoma multiforme (GBM) is limited by resistance. The goal of this study was to elucidate mechanisms of temozolomide resistance in GBM. Experimental Design: We developed an in vivoGBMmodel of temozolomide resistance and used paired parental and temozolomide-resistant tumors to define the mechanisms underlying the development of resistance and the influence of histone deacetylation (HDAC) inhibition. Results: Analysis of paired parental and resistant lines showed upregulation of O6-methylguanine-DNA methyltransferase (MGMT) expression in 3 of the 5 resistant xenografts. While no significant change was detected in MGMT promoter methylation between parental and derivative-resistant samples, chromatin immunoprecipitation showed an association between MGMT upregulation and elevated acetylation of lysine 9 of histone H3 (H3K9-ac) and decreased dimethylation (H3K9-me2) in GBM12 and GBM14. In contrast, temozolomide resistance development in GBM22 was not linked to MGMT expression, and both parental and resistant lines had low H3K9-ac and high H3K9-me2 within the MGMT promoter. In the GBM12TMZ-resistant line, MGMT reexpression was accompanied by increased recruitment of SP1, C-JUN, NF-kB, and p300 within the MGMT promoter. Interestingly, combined treatment of GBM12 flank xenografts with temozolomide and the HDAC inhibitor suberoylanilide hydroxamic acid (SAHA) favored the evolution of temozolomide resistance by MGMT overexpression as compared with treatment with temozolomide alone. Conclusion: This study shows, for the first time, a unique mechanism of temozolomide resistance development driven by chromatin-mediated MGMT upregulation and highlights the potential for epigenetically directed therapies to influence the mechanisms of resistance development in GBM.

AB - Purpose: The therapeutic benefit of temozolomide in glioblastoma multiforme (GBM) is limited by resistance. The goal of this study was to elucidate mechanisms of temozolomide resistance in GBM. Experimental Design: We developed an in vivoGBMmodel of temozolomide resistance and used paired parental and temozolomide-resistant tumors to define the mechanisms underlying the development of resistance and the influence of histone deacetylation (HDAC) inhibition. Results: Analysis of paired parental and resistant lines showed upregulation of O6-methylguanine-DNA methyltransferase (MGMT) expression in 3 of the 5 resistant xenografts. While no significant change was detected in MGMT promoter methylation between parental and derivative-resistant samples, chromatin immunoprecipitation showed an association between MGMT upregulation and elevated acetylation of lysine 9 of histone H3 (H3K9-ac) and decreased dimethylation (H3K9-me2) in GBM12 and GBM14. In contrast, temozolomide resistance development in GBM22 was not linked to MGMT expression, and both parental and resistant lines had low H3K9-ac and high H3K9-me2 within the MGMT promoter. In the GBM12TMZ-resistant line, MGMT reexpression was accompanied by increased recruitment of SP1, C-JUN, NF-kB, and p300 within the MGMT promoter. Interestingly, combined treatment of GBM12 flank xenografts with temozolomide and the HDAC inhibitor suberoylanilide hydroxamic acid (SAHA) favored the evolution of temozolomide resistance by MGMT overexpression as compared with treatment with temozolomide alone. Conclusion: This study shows, for the first time, a unique mechanism of temozolomide resistance development driven by chromatin-mediated MGMT upregulation and highlights the potential for epigenetically directed therapies to influence the mechanisms of resistance development in GBM.

UR - http://www.scopus.com/inward/record.url?scp=84864495476&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84864495476&partnerID=8YFLogxK

U2 - 10.1158/1078-0432.CCR-12-0560

DO - 10.1158/1078-0432.CCR-12-0560

M3 - Article

VL - 18

SP - 4070

EP - 4079

JO - Clinical Cancer Research

JF - Clinical Cancer Research

SN - 1078-0432

IS - 15

ER -