TY - JOUR
T1 - MAPK pathway activation leads to Bim loss and histone deacetylase inhibitor resistance
T2 - Rationale to combine romidepsin with an MEK inhibitor
AU - Chakraborty, Arup R.
AU - Robey, Robert W.
AU - Luchenko, Victoria L.
AU - Zhan, Zhirong
AU - Piekarz, Richard L.
AU - Gillet, Jean Pierre
AU - Kossenkov, Andrew V.
AU - Wilkerson, Julia
AU - Showe, Louise C.
AU - Gottesman, Michael M.
AU - Collie, Nathan L.
AU - Bates, Susan E.
N1 - Funding Information:
The authors gratefully acknowledge Donna Butcher and Dr Miriam R. Anver of the Histotechnology Laboratory at the Frederick National Laboratory for Cancer Research for the immunohistochemistry analyses. This work was supported in part by the Intramural Research Program of the National Institutes of Health, National Cancer Institute, Center for Cancer Research and by a Cooperative Research and Development Agreement with Celgene Corporation, as well as by NCI grant RO1 CA132098 (to L.C.S.). The Wistar Institute Genomics and Bioinformatics facilities are supported by National Institutes of Health, Cancer Center Support Grant P30 CA010815. The content of this publication does not necessarily reflect the views or policies of the Department of Health and Human Services, nor does mention of trade names, commercial products, or organizations imply endorsement by the U.S. Government. Conflict-of-interest disclosure: This research was supported in part by a cooperative research and development agreement between the National Cancer Institute and Celgene Corporation (S.E.B.). The remaining authors declare no competing financial interests.
Funding Information:
1Department of Biological Sciences, Texas Tech University, Lubbock, TX; 2Medical Oncology Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD; 3Cancer Therapy Evaluation Program, Division of Cancer Treatment and Diagnosis, National Cancer Institute, National Institutes of Health, Rockville, MD; 4Laboratory of Cell Biology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD; 5The Wistar Institute, Philadelphia, PA
Funding Information:
Conflict-of-interest disclosure: This research was supported in part by a cooperative research and development agreement between the National Cancer Institute and Celgene Corporation (S.E.B.). The remaining authors declare no competing financial interests.
Funding Information:
This work was supported in part by the Intramural Research Program of the National Institutes of Health, National Cancer Institute, Center for Cancer Research and by a Cooperative Research and Development Agreement with Celgene Corporation, as well as by NCI grant RO1 CA132098 (to L.C.S.). The Wistar Institute
Funding Information:
Correspondence: Susan E. Bates, National Cancer Institute, National Institutes of Health, 9000 Rockville Pike, Bldg 10, Room 12N226, Bethesda, MD 20892; e-mail: batess@helix.nih.gov.
Publisher Copyright:
© 2013 by The American Society of Hematology.
PY - 2013/5/16
Y1 - 2013/5/16
N2 - To identify molecular determinants of histone deacetylase inhibitor (HDI) resistance, we selected HuT78 cutaneous T-cell lymphoma (CTCL) cells with romidepsin in the presence of P-glycoprotein inhibitors to prevent transporter upregulation. Resistant sublines were 250- to 385-fold resistant to romidepsin and were resistant to apoptosis induced by apicidin, entinostat, panobinostat, belinostat, and vorinostat. A custom TaqMan array identified increased insulin receptor (INSR) gene expression; immunoblot analysis confirmed increased protein expression and a four- to eightfold increase in mitogen-activated protein kinase (MAPK) kinase (MEK) phosphorylation in resistant cells compared with parental cells. Resistant cells were exquisitely sensitive to MEK inhibitors, and apoptosis correlated with restoration of proapoptotic Bim. Romidepsin combined with MEK inhibitors yielded greater apoptosis in cells expressing mutant KRAS compared with romidepsin treatment alone. Gene expression analysis of samples obtained from patients with CTCL enrolled on the NCI1312 phase 2 study of romidepsin in T-cell lymphoma suggested perturbation of the MAPK pathway by romidepsin. Immunohistochemical analysis of Bim expression demonstrated decreased expression in some skin biopsies at disease progression. These findings implicate increased activation of MEK and decreased Bim expression as a resistance mechanism to HDIs, supporting combination of romidepsin with MEK inhibitors in clinical trials.
AB - To identify molecular determinants of histone deacetylase inhibitor (HDI) resistance, we selected HuT78 cutaneous T-cell lymphoma (CTCL) cells with romidepsin in the presence of P-glycoprotein inhibitors to prevent transporter upregulation. Resistant sublines were 250- to 385-fold resistant to romidepsin and were resistant to apoptosis induced by apicidin, entinostat, panobinostat, belinostat, and vorinostat. A custom TaqMan array identified increased insulin receptor (INSR) gene expression; immunoblot analysis confirmed increased protein expression and a four- to eightfold increase in mitogen-activated protein kinase (MAPK) kinase (MEK) phosphorylation in resistant cells compared with parental cells. Resistant cells were exquisitely sensitive to MEK inhibitors, and apoptosis correlated with restoration of proapoptotic Bim. Romidepsin combined with MEK inhibitors yielded greater apoptosis in cells expressing mutant KRAS compared with romidepsin treatment alone. Gene expression analysis of samples obtained from patients with CTCL enrolled on the NCI1312 phase 2 study of romidepsin in T-cell lymphoma suggested perturbation of the MAPK pathway by romidepsin. Immunohistochemical analysis of Bim expression demonstrated decreased expression in some skin biopsies at disease progression. These findings implicate increased activation of MEK and decreased Bim expression as a resistance mechanism to HDIs, supporting combination of romidepsin with MEK inhibitors in clinical trials.
UR - http://www.scopus.com/inward/record.url?scp=84881001025&partnerID=8YFLogxK
U2 - 10.1182/blood-2012-08-449140
DO - 10.1182/blood-2012-08-449140
M3 - Article
C2 - 23532732
AN - SCOPUS:84881001025
VL - 121
SP - 4115
EP - 4125
JO - Blood
JF - Blood
SN - 0006-4971
IS - 20
ER -