FBW7 mutations in leukemic cells mediate NOTCH pathway activation and resistance to γ-secretase inhibitors

Jennifer O'Neil, Jonathan Grim, Peter Strack, Sudhir Rao, Deanne Tibbitts, Christopher Winter, James Hardwick, Markus Welcker, Jules P. Meijerink, Rob Pieters, Giulio Draetta, Rosalie Sears, Bruce E. Clurman, A. Thomas Look

Research output: Contribution to journalArticle

431 Citations (Scopus)

Abstract

γ-secretase inhibitors (GSIs) can block NOTCH receptor signaling in vitro and therefore offer an attractive targeted therapy for tumors dependent on deregulated NOTCH activity. To clarify the basis for GSI resistance in T cell acute lymphoblastic leukemia (T-ALL), we studied T-ALL cell lines with constitutive expression of the NOTCH intracellular domain (NICD), but that lacked C-terminal truncating mutations in NOTCH1. Each of the seven cell lines examined and 7 of 81 (8.6%) primary T-ALL samples harbored either a mutation or homozygous deletion of the gene FBW7, a ubiquitin ligase implicated in NICD turnover. Indeed, we show that FBW7 mutants cannot bind to the NICD and define the phosphodegron region of the NICD required for FBW7 binding. Although the mutant forms of FBW7 were still able to bind to MYC, they do not target it for degradation, suggesting that stabilization of both NICD and its principle downstream target, MYC, may contribute to transformation in leukemias with FBW7 mutations. In addition, we show that all seven leukemic cell lines with FBW7 mutations were resistant to the MRK-003 GSI. Most of these resistant lines also failed to down-regulate the mRNA levels of the NOTCH targets MYC and DELTEX1 after treatment with MRK-003, implying that residual NOTCH signaling in T-ALLs with FBW7 mutations contributes to GSI resistance. JEM

Original languageEnglish (US)
Pages (from-to)1813-1824
Number of pages12
JournalJournal of Experimental Medicine
Volume204
Issue number8
DOIs
StatePublished - Aug 6 2007

Fingerprint

Amyloid Precursor Protein Secretases
Mutation
Precursor T-Cell Lymphoblastic Leukemia-Lymphoma
Cell Line
T-Cell Leukemia
Gene Deletion
Ligases
Ubiquitin
Precursor Cell Lymphoblastic Leukemia-Lymphoma
Leukemia
Down-Regulation
T-Lymphocytes
Messenger RNA
Neoplasms

ASJC Scopus subject areas

  • Immunology

Cite this

FBW7 mutations in leukemic cells mediate NOTCH pathway activation and resistance to γ-secretase inhibitors. / O'Neil, Jennifer; Grim, Jonathan; Strack, Peter; Rao, Sudhir; Tibbitts, Deanne; Winter, Christopher; Hardwick, James; Welcker, Markus; Meijerink, Jules P.; Pieters, Rob; Draetta, Giulio; Sears, Rosalie; Clurman, Bruce E.; Look, A. Thomas.

In: Journal of Experimental Medicine, Vol. 204, No. 8, 06.08.2007, p. 1813-1824.

Research output: Contribution to journalArticle

O'Neil, J, Grim, J, Strack, P, Rao, S, Tibbitts, D, Winter, C, Hardwick, J, Welcker, M, Meijerink, JP, Pieters, R, Draetta, G, Sears, R, Clurman, BE & Look, AT 2007, 'FBW7 mutations in leukemic cells mediate NOTCH pathway activation and resistance to γ-secretase inhibitors', Journal of Experimental Medicine, vol. 204, no. 8, pp. 1813-1824. https://doi.org/10.1084/jem.20070876
O'Neil, Jennifer ; Grim, Jonathan ; Strack, Peter ; Rao, Sudhir ; Tibbitts, Deanne ; Winter, Christopher ; Hardwick, James ; Welcker, Markus ; Meijerink, Jules P. ; Pieters, Rob ; Draetta, Giulio ; Sears, Rosalie ; Clurman, Bruce E. ; Look, A. Thomas. / FBW7 mutations in leukemic cells mediate NOTCH pathway activation and resistance to γ-secretase inhibitors. In: Journal of Experimental Medicine. 2007 ; Vol. 204, No. 8. pp. 1813-1824.
@article{23b39558a7f24831a22508f9c8f1ab86,
title = "FBW7 mutations in leukemic cells mediate NOTCH pathway activation and resistance to γ-secretase inhibitors",
abstract = "γ-secretase inhibitors (GSIs) can block NOTCH receptor signaling in vitro and therefore offer an attractive targeted therapy for tumors dependent on deregulated NOTCH activity. To clarify the basis for GSI resistance in T cell acute lymphoblastic leukemia (T-ALL), we studied T-ALL cell lines with constitutive expression of the NOTCH intracellular domain (NICD), but that lacked C-terminal truncating mutations in NOTCH1. Each of the seven cell lines examined and 7 of 81 (8.6{\%}) primary T-ALL samples harbored either a mutation or homozygous deletion of the gene FBW7, a ubiquitin ligase implicated in NICD turnover. Indeed, we show that FBW7 mutants cannot bind to the NICD and define the phosphodegron region of the NICD required for FBW7 binding. Although the mutant forms of FBW7 were still able to bind to MYC, they do not target it for degradation, suggesting that stabilization of both NICD and its principle downstream target, MYC, may contribute to transformation in leukemias with FBW7 mutations. In addition, we show that all seven leukemic cell lines with FBW7 mutations were resistant to the MRK-003 GSI. Most of these resistant lines also failed to down-regulate the mRNA levels of the NOTCH targets MYC and DELTEX1 after treatment with MRK-003, implying that residual NOTCH signaling in T-ALLs with FBW7 mutations contributes to GSI resistance. JEM",
author = "Jennifer O'Neil and Jonathan Grim and Peter Strack and Sudhir Rao and Deanne Tibbitts and Christopher Winter and James Hardwick and Markus Welcker and Meijerink, {Jules P.} and Rob Pieters and Giulio Draetta and Rosalie Sears and Clurman, {Bruce E.} and Look, {A. Thomas}",
year = "2007",
month = "8",
day = "6",
doi = "10.1084/jem.20070876",
language = "English (US)",
volume = "204",
pages = "1813--1824",
journal = "Journal of Experimental Medicine",
issn = "0022-1007",
publisher = "Rockefeller University Press",
number = "8",

}

TY - JOUR

T1 - FBW7 mutations in leukemic cells mediate NOTCH pathway activation and resistance to γ-secretase inhibitors

AU - O'Neil, Jennifer

AU - Grim, Jonathan

AU - Strack, Peter

AU - Rao, Sudhir

AU - Tibbitts, Deanne

AU - Winter, Christopher

AU - Hardwick, James

AU - Welcker, Markus

AU - Meijerink, Jules P.

AU - Pieters, Rob

AU - Draetta, Giulio

AU - Sears, Rosalie

AU - Clurman, Bruce E.

AU - Look, A. Thomas

PY - 2007/8/6

Y1 - 2007/8/6

N2 - γ-secretase inhibitors (GSIs) can block NOTCH receptor signaling in vitro and therefore offer an attractive targeted therapy for tumors dependent on deregulated NOTCH activity. To clarify the basis for GSI resistance in T cell acute lymphoblastic leukemia (T-ALL), we studied T-ALL cell lines with constitutive expression of the NOTCH intracellular domain (NICD), but that lacked C-terminal truncating mutations in NOTCH1. Each of the seven cell lines examined and 7 of 81 (8.6%) primary T-ALL samples harbored either a mutation or homozygous deletion of the gene FBW7, a ubiquitin ligase implicated in NICD turnover. Indeed, we show that FBW7 mutants cannot bind to the NICD and define the phosphodegron region of the NICD required for FBW7 binding. Although the mutant forms of FBW7 were still able to bind to MYC, they do not target it for degradation, suggesting that stabilization of both NICD and its principle downstream target, MYC, may contribute to transformation in leukemias with FBW7 mutations. In addition, we show that all seven leukemic cell lines with FBW7 mutations were resistant to the MRK-003 GSI. Most of these resistant lines also failed to down-regulate the mRNA levels of the NOTCH targets MYC and DELTEX1 after treatment with MRK-003, implying that residual NOTCH signaling in T-ALLs with FBW7 mutations contributes to GSI resistance. JEM

AB - γ-secretase inhibitors (GSIs) can block NOTCH receptor signaling in vitro and therefore offer an attractive targeted therapy for tumors dependent on deregulated NOTCH activity. To clarify the basis for GSI resistance in T cell acute lymphoblastic leukemia (T-ALL), we studied T-ALL cell lines with constitutive expression of the NOTCH intracellular domain (NICD), but that lacked C-terminal truncating mutations in NOTCH1. Each of the seven cell lines examined and 7 of 81 (8.6%) primary T-ALL samples harbored either a mutation or homozygous deletion of the gene FBW7, a ubiquitin ligase implicated in NICD turnover. Indeed, we show that FBW7 mutants cannot bind to the NICD and define the phosphodegron region of the NICD required for FBW7 binding. Although the mutant forms of FBW7 were still able to bind to MYC, they do not target it for degradation, suggesting that stabilization of both NICD and its principle downstream target, MYC, may contribute to transformation in leukemias with FBW7 mutations. In addition, we show that all seven leukemic cell lines with FBW7 mutations were resistant to the MRK-003 GSI. Most of these resistant lines also failed to down-regulate the mRNA levels of the NOTCH targets MYC and DELTEX1 after treatment with MRK-003, implying that residual NOTCH signaling in T-ALLs with FBW7 mutations contributes to GSI resistance. JEM

UR - http://www.scopus.com/inward/record.url?scp=34547780475&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=34547780475&partnerID=8YFLogxK

U2 - 10.1084/jem.20070876

DO - 10.1084/jem.20070876

M3 - Article

VL - 204

SP - 1813

EP - 1824

JO - Journal of Experimental Medicine

JF - Journal of Experimental Medicine

SN - 0022-1007

IS - 8

ER -