Methylthioadenosine phosphorylase as target for chemoselective treatment of T-cell acute lymphoblastic leukemic cells

Autoren

Thomas Efferth
Hayato Miyachi
Hans Günter Drexler
Erich Gebhart

DOI

10.1006/bcmd.2002.0483

ISSN

1079-9796

Ausgabe der Veröffentlichung

1

Zeitschrift

Blood Cells, Molecules, and Diseases

Sprache

en

Paginierung

47 - 56

Datum der Veröffentlichung

2002

Status

Published

Herausgeber

Elsevier BV

Herausgeber URL

http://dx.doi.org/10.1006/bcmd.2002.0483

Datum der Datenerfassung

2023

Titel

Methylthioadenosine Phosphorylase as Target for Chemoselective Treatment of T-Cell Acute Lymphoblastic Leukemic Cells

Ausgabe der Zeitschrift

28

Data source: Crossref

Abstract

We analyzed the role of methylthioadenosine phosphorylase (MTAP) for chemoselective treatment of T-cell acute lymphoblastic leukemia (T-ALL). MTAP converts methylthioadenosine into adenine which serves as an alternative purine source, if de novo purine biosynthesis is inhibited by antimetabolites (i.e., methotrexate). The idea of the chemoselectivity concept is that tumors with MTAP deletion at chromosome 9p21 are more susceptible to antimetabolites than normal cells without such a deletion. First, we screened 13 T-ALL lines for 9p21 deletions by comparative genomic hybridization. Five cell lines revealed deletions at the short arm of chromosome 9, dim(9p21pter). Further analyses were performed with CEM cells in which the 9p21 deletion was corroborated by fluorescence in situ hybridization. CEM cells were transfected with an MTAP expression vector. A green fluorescent protein (GFP) plasmid was cotransfected, to monitor the transfection efficacy by flow cytometry. The response of MTAP-transfected cells to the antimetabolites methotrexate (MTX), trimetrexate (TMX), and L-alanosine (ALA) was decreased compared to mock control transfectants using growth inhibition assays. The activity of doxorubicin (DOX) which is not involved in DNA biosynthesis was not changed in MTAP transfectants. As the p16(INK4a) tumor suppressor gene resides also at 9p21, we transfected CEM cells with a p16(INK4a) expression vector. These transfectant cells were more resistant to all four drugs indicating that p16(INK4a) did not specifically affect antimetabolites. The chemoselective effect of antimetabolites in MTAP-deleted tumor cells may, however, be compensated by the development of drug resistance. To prove this possibility, we analyzed an MTX-resistant subline, CEM/MTX1500LV, in which the MTX-resistance conferring dihydrofolate reductase (DHFR) gene was amplified. While TMX exhibited considerable cross-resistance in CEM/MTX1500LV cells, ALA did not. Thus, ALA could exhibit chemoselectivity in 9p21/MTAP-deleted cells, even if DHFR amplification occurs. We conclude that ALA may be more suitable than MTX or TMX for MTAP-mediated chemoselective treatment of T-ALL. Pretherapeutical detection of 9p21 and MTAP deletion may be helpful in developing a predictive molecular chemosensitivity test for T-ALL.

Addresses

Virtual Campus Rhineland-Palatinate, Mainz, Germany. efferth@vcrp.de

Autoren

Thomas Efferth
Thomas Efferth
Hayato Miyachi
Hans Günther Drexler
Erich Gebhart

DOI

10.1006/bcmd.2002.0483

eISSN

1096-0961

Externe Identifier

PubMed Identifier: 11987241

Open access

false

ISSN

1079-9796

Ausgabe der Veröffentlichung

1

Zeitschrift

Blood cells, molecules & diseases

Schlüsselwörter

Tumor Cells, Cultured
Chromosomes, Human, Pair 9
Humans
Chromosome Deletion
Trimetrexate
Purine-Nucleoside Phosphorylase
Alanine
Neoplasm Proteins
Antimetabolites, Antineoplastic
Transfection
Cell Division
Drug Resistance, Neoplasm
Cyclin-Dependent Kinase Inhibitor p16
Leukemia-Lymphoma, Adult T-Cell

Sprache

eng

Medium

Print

Paginierung

47 - 56

Datum der Veröffentlichung

2002

Status

Published

Datum der Datenerfassung

2002

Titel

Methylthioadenosine phosphorylase as target for chemoselective treatment of T-cell acute lymphoblastic leukemic cells.

Sub types

Journal Article

Ausgabe der Zeitschrift

28

Data source: Europe PubMed Central

Abstract

We analyzed the role of methylthioadenosine phosphorylase (MTAP) for chemoselective treatment of T-cell acute lymphoblastic leukemia (T-ALL). MTAP converts methylthioadenosine into adenine which serves as an alternative purine source, if de novo purine biosynthesis is inhibited by antimetabolites (i.e., methotrexate). The idea of the chemoselectivity concept is that tumors with MTAP deletion at chromosome 9p21 are more susceptible to antimetabolites than normal cells without such a deletion. First, we screened 13 T-ALL lines for 9p21 deletions by comparative genomic hybridization. Five cell lines revealed deletions at the short arm of chromosome 9, dim(9p21pter). Further analyses were performed with CEM cells in which the 9p21 deletion was corroborated by fluorescence in situ hybridization. CEM cells were transfected with an MTAP expression vector. A green fluorescent protein (GFP) plasmid was cotransfected, to monitor the transfection efficacy by flow cytometry. The response of MTAP-transfected cells to the antimetabolites methotrexate (MTX), trimetrexate (TMX), and L-alanosine (ALA) was decreased compared to mock control transfectants using growth inhibition assays. The activity of doxorubicin (DOX) which is not involved in DNA biosynthesis was not changed in MTAP transfectants. As the p16(INK4a) tumor suppressor gene resides also at 9p21, we transfected CEM cells with a p16(INK4a) expression vector. These transfectant cells were more resistant to all four drugs indicating that p16(INK4a) did not specifically affect antimetabolites. The chemoselective effect of antimetabolites in MTAP-deleted tumor cells may, however, be compensated by the development of drug resistance. To prove this possibility, we analyzed an MTX-resistant subline, CEM/MTX1500LV, in which the MTX-resistance conferring dihydrofolate reductase (DHFR) gene was amplified. While TMX exhibited considerable cross-resistance in CEM/MTX1500LV cells, ALA did not. Thus, ALA could exhibit chemoselectivity in 9p21/MTAP-deleted cells, even if DHFR amplification occurs. We conclude that ALA may be more suitable than MTX or TMX for MTAP-mediated chemoselective treatment of T-ALL. Pretherapeutical detection of 9p21 and MTAP deletion may be helpful in developing a predictive molecular chemosensitivity test for T-ALL.

Autoren

Thomas Efferth
Hayato Miyachi
Hans Günther Drexler
Erich Gebhart

Autoren-URL

https://www.ncbi.nlm.nih.gov/pubmed/11987241

DOI

10.1006/bcmd.2002.0483

ISSN

1079-9796

Ausgabe der Veröffentlichung

1

Zeitschrift

Blood Cells Mol Dis

Schlüsselwörter

Alanine
Antimetabolites, Antineoplastic
Cell Division
Chromosome Deletion
Chromosomes, Human, Pair 9
Cyclin-Dependent Kinase Inhibitor p16
Drug Resistance, Neoplasm
Humans
Leukemia-Lymphoma, Adult T-Cell
Neoplasm Proteins
Purine-Nucleoside Phosphorylase
Transfection
Trimetrexate
Tumor Cells, Cultured

Sprache

eng

Country

United States

Paginierung

47 - 56

PII

S1079979602904833

Datum der Veröffentlichung

2002

Status

Published

Datum, an dem der Datensatz öffentlich gemacht wurde

2003

Titel

Methylthioadenosine phosphorylase as target for chemoselective treatment of T-cell acute lymphoblastic leukemic cells.

Sub types

Journal Article

Ausgabe der Zeitschrift

28

Data source: PubMed

Methylthioadenosine phosphorylase as target for chemoselective treatment of T-cell acute lymphoblastic leukemic cells

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