Advanced

NUDT15 Hydrolyzes 6-Thio-DeoxyGTP to Mediate the Anticancer Efficacy of 6-Thioguanine

Valerie, Nicholas C K; Hagenkort, Anna; Page, Brent D G; Masuyer, Geoffrey; Rehling, Daniel; Carter, Megan; Bevc, Luka; Herr, Patrick; Homan, Evert and Sheppard, Nina G, et al. (2016) In Cancer Research 76(18). p.11-5501
Abstract

Thiopurines are a standard treatment for childhood leukemia, but like all chemotherapeutics, their use is limited by inherent or acquired resistance in patients. Recently, the nucleoside diphosphate hydrolase NUDT15 has received attention on the basis of its ability to hydrolyze the thiopurine effector metabolites 6-thio-deoxyGTP (6-thio-dGTP) and 6-thio-GTP, thereby limiting the efficacy of thiopurines. In particular, increasing evidence suggests an association between the NUDT15 missense variant, R139C, and thiopurine sensitivity. In this study, we elucidated the role of NUDT15 and NUDT15 R139C in thiopurine metabolism. In vitro and cellular results argued that 6-thio-dGTP and 6-thio-GTP are favored substrates for NUDT15, a finding... (More)

Thiopurines are a standard treatment for childhood leukemia, but like all chemotherapeutics, their use is limited by inherent or acquired resistance in patients. Recently, the nucleoside diphosphate hydrolase NUDT15 has received attention on the basis of its ability to hydrolyze the thiopurine effector metabolites 6-thio-deoxyGTP (6-thio-dGTP) and 6-thio-GTP, thereby limiting the efficacy of thiopurines. In particular, increasing evidence suggests an association between the NUDT15 missense variant, R139C, and thiopurine sensitivity. In this study, we elucidated the role of NUDT15 and NUDT15 R139C in thiopurine metabolism. In vitro and cellular results argued that 6-thio-dGTP and 6-thio-GTP are favored substrates for NUDT15, a finding supported by a crystallographic determination of NUDT15 in complex with 6-thio-GMP. We found that NUDT15 R139C mutation did not affect enzymatic activity but instead negatively influenced protein stability, likely due to a loss of supportive intramolecular bonds that caused rapid proteasomal degradation in cells. Mechanistic investigations in cells indicated that NUDT15 ablation potentiated induction of the DNA damage checkpoint and cancer cell death by 6-thioguanine. Taken together, our results defined how NUDT15 limits thiopurine efficacy and how genetic ablation via the R139C missense mutation confers sensitivity to thiopurine treatment in patients. Cancer Res; 76(18); 5501-11. ©2016 AACR.

(Less)
Please use this url to cite or link to this publication:
author
, et al. (More)
(Less)
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Antimetabolites, Antineoplastic/metabolism, Blotting, Western, Cell Line, Tumor, Chromatography, High Pressure Liquid, Crystallography, X-Ray, Humans, Mutation, Missense, Protein Stability, Pyrophosphatases/chemistry, Thioguanine/metabolism
in
Cancer Research
volume
76
issue
18
pages
11 pages
publisher
American Association for Cancer Research Inc.
external identifiers
  • scopus:84988944149
ISSN
1538-7445
DOI
10.1158/0008-5472.CAN-16-0584
language
English
LU publication?
no
id
a08d86cd-9fe1-4261-ab19-480e8606734a
date added to LUP
2019-05-07 08:12:19
date last changed
2019-10-15 07:03:00
@article{a08d86cd-9fe1-4261-ab19-480e8606734a,
  abstract     = {<p>Thiopurines are a standard treatment for childhood leukemia, but like all chemotherapeutics, their use is limited by inherent or acquired resistance in patients. Recently, the nucleoside diphosphate hydrolase NUDT15 has received attention on the basis of its ability to hydrolyze the thiopurine effector metabolites 6-thio-deoxyGTP (6-thio-dGTP) and 6-thio-GTP, thereby limiting the efficacy of thiopurines. In particular, increasing evidence suggests an association between the NUDT15 missense variant, R139C, and thiopurine sensitivity. In this study, we elucidated the role of NUDT15 and NUDT15 R139C in thiopurine metabolism. In vitro and cellular results argued that 6-thio-dGTP and 6-thio-GTP are favored substrates for NUDT15, a finding supported by a crystallographic determination of NUDT15 in complex with 6-thio-GMP. We found that NUDT15 R139C mutation did not affect enzymatic activity but instead negatively influenced protein stability, likely due to a loss of supportive intramolecular bonds that caused rapid proteasomal degradation in cells. Mechanistic investigations in cells indicated that NUDT15 ablation potentiated induction of the DNA damage checkpoint and cancer cell death by 6-thioguanine. Taken together, our results defined how NUDT15 limits thiopurine efficacy and how genetic ablation via the R139C missense mutation confers sensitivity to thiopurine treatment in patients. Cancer Res; 76(18); 5501-11. ©2016 AACR.</p>},
  author       = {Valerie, Nicholas C K and Hagenkort, Anna and Page, Brent D G and Masuyer, Geoffrey and Rehling, Daniel and Carter, Megan and Bevc, Luka and Herr, Patrick and Homan, Evert and Sheppard, Nina G and Stenmark, Pål and Jemth, Ann-Sofie and Helleday, Thomas},
  issn         = {1538-7445},
  keyword      = {Antimetabolites, Antineoplastic/metabolism,Blotting, Western,Cell Line, Tumor,Chromatography, High Pressure Liquid,Crystallography, X-Ray,Humans,Mutation, Missense,Protein Stability,Pyrophosphatases/chemistry,Thioguanine/metabolism},
  language     = {eng},
  month        = {09},
  number       = {18},
  pages        = {11--5501},
  publisher    = {American Association for Cancer Research Inc.},
  series       = {Cancer Research},
  title        = {NUDT15 Hydrolyzes 6-Thio-DeoxyGTP to Mediate the Anticancer Efficacy of 6-Thioguanine},
  url          = {http://dx.doi.org/10.1158/0008-5472.CAN-16-0584},
  volume       = {76},
  year         = {2016},
}