Cargando…
Crystal structures of NUDT15 variants enabled by a potent inhibitor reveal the structural basis for thiopurine sensitivity
The enzyme NUDT15 efficiently hydrolyzes the active metabolites of thiopurine drugs, which are routinely used for treating cancer and inflammatory diseases. Loss-of-function variants in NUDT15 are strongly associated with thiopurine intolerance, such as leukopenia, and preemptive NUDT15 genotyping h...
Autores principales: | , , , , , , , , , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society for Biochemistry and Molecular Biology
2021
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8079283/ https://www.ncbi.nlm.nih.gov/pubmed/33753169 http://dx.doi.org/10.1016/j.jbc.2021.100568 |
_version_ | 1783685194673291264 |
---|---|
author | Rehling, Daniel Zhang, Si Min Jemth, Ann-Sofie Koolmeister, Tobias Throup, Adam Wallner, Olov Scaletti, Emma Moriyama, Takaya Nishii, Rina Davies, Jonathan Desroses, Matthieu Rudd, Sean G. Scobie, Martin Homan, Evert Berglund, Ulrika Warpman Yang, Jun J. Helleday, Thomas Stenmark, Pål |
author_facet | Rehling, Daniel Zhang, Si Min Jemth, Ann-Sofie Koolmeister, Tobias Throup, Adam Wallner, Olov Scaletti, Emma Moriyama, Takaya Nishii, Rina Davies, Jonathan Desroses, Matthieu Rudd, Sean G. Scobie, Martin Homan, Evert Berglund, Ulrika Warpman Yang, Jun J. Helleday, Thomas Stenmark, Pål |
author_sort | Rehling, Daniel |
collection | PubMed |
description | The enzyme NUDT15 efficiently hydrolyzes the active metabolites of thiopurine drugs, which are routinely used for treating cancer and inflammatory diseases. Loss-of-function variants in NUDT15 are strongly associated with thiopurine intolerance, such as leukopenia, and preemptive NUDT15 genotyping has been clinically implemented to personalize thiopurine dosing. However, understanding the molecular consequences of these variants has been difficult, as no structural information was available for NUDT15 proteins encoded by clinically actionable pharmacogenetic variants because of their inherent instability. Recently, the small molecule NUDT15 inhibitor TH1760 has been shown to sensitize cells to thiopurines, through enhanced accumulation of 6-thio-guanine in DNA. Building upon this, we herein report the development of the potent and specific NUDT15 inhibitor, TH7755. TH7755 demonstrates a greatly improved cellular target engagement and 6-thioguanine potentiation compared with TH1760, while showing no cytotoxicity on its own. This potent inhibitor also stabilized NUDT15, enabling analysis by X-ray crystallography. We have determined high-resolution structures of the clinically relevant NUDT15 variants Arg139Cys, Arg139His, Val18Ile, and V18_V19insGlyVal. These structures provide clear insights into the structural basis for the thiopurine intolerance phenotype observed in patients carrying these pharmacogenetic variants. These findings will aid in predicting the effects of new NUDT15 sequence variations yet to be discovered in the clinic. |
format | Online Article Text |
id | pubmed-8079283 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | American Society for Biochemistry and Molecular Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-80792832021-04-30 Crystal structures of NUDT15 variants enabled by a potent inhibitor reveal the structural basis for thiopurine sensitivity Rehling, Daniel Zhang, Si Min Jemth, Ann-Sofie Koolmeister, Tobias Throup, Adam Wallner, Olov Scaletti, Emma Moriyama, Takaya Nishii, Rina Davies, Jonathan Desroses, Matthieu Rudd, Sean G. Scobie, Martin Homan, Evert Berglund, Ulrika Warpman Yang, Jun J. Helleday, Thomas Stenmark, Pål J Biol Chem Research Article The enzyme NUDT15 efficiently hydrolyzes the active metabolites of thiopurine drugs, which are routinely used for treating cancer and inflammatory diseases. Loss-of-function variants in NUDT15 are strongly associated with thiopurine intolerance, such as leukopenia, and preemptive NUDT15 genotyping has been clinically implemented to personalize thiopurine dosing. However, understanding the molecular consequences of these variants has been difficult, as no structural information was available for NUDT15 proteins encoded by clinically actionable pharmacogenetic variants because of their inherent instability. Recently, the small molecule NUDT15 inhibitor TH1760 has been shown to sensitize cells to thiopurines, through enhanced accumulation of 6-thio-guanine in DNA. Building upon this, we herein report the development of the potent and specific NUDT15 inhibitor, TH7755. TH7755 demonstrates a greatly improved cellular target engagement and 6-thioguanine potentiation compared with TH1760, while showing no cytotoxicity on its own. This potent inhibitor also stabilized NUDT15, enabling analysis by X-ray crystallography. We have determined high-resolution structures of the clinically relevant NUDT15 variants Arg139Cys, Arg139His, Val18Ile, and V18_V19insGlyVal. These structures provide clear insights into the structural basis for the thiopurine intolerance phenotype observed in patients carrying these pharmacogenetic variants. These findings will aid in predicting the effects of new NUDT15 sequence variations yet to be discovered in the clinic. American Society for Biochemistry and Molecular Biology 2021-03-19 /pmc/articles/PMC8079283/ /pubmed/33753169 http://dx.doi.org/10.1016/j.jbc.2021.100568 Text en © 2021 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Research Article Rehling, Daniel Zhang, Si Min Jemth, Ann-Sofie Koolmeister, Tobias Throup, Adam Wallner, Olov Scaletti, Emma Moriyama, Takaya Nishii, Rina Davies, Jonathan Desroses, Matthieu Rudd, Sean G. Scobie, Martin Homan, Evert Berglund, Ulrika Warpman Yang, Jun J. Helleday, Thomas Stenmark, Pål Crystal structures of NUDT15 variants enabled by a potent inhibitor reveal the structural basis for thiopurine sensitivity |
title | Crystal structures of NUDT15 variants enabled by a potent inhibitor reveal the structural basis for thiopurine sensitivity |
title_full | Crystal structures of NUDT15 variants enabled by a potent inhibitor reveal the structural basis for thiopurine sensitivity |
title_fullStr | Crystal structures of NUDT15 variants enabled by a potent inhibitor reveal the structural basis for thiopurine sensitivity |
title_full_unstemmed | Crystal structures of NUDT15 variants enabled by a potent inhibitor reveal the structural basis for thiopurine sensitivity |
title_short | Crystal structures of NUDT15 variants enabled by a potent inhibitor reveal the structural basis for thiopurine sensitivity |
title_sort | crystal structures of nudt15 variants enabled by a potent inhibitor reveal the structural basis for thiopurine sensitivity |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8079283/ https://www.ncbi.nlm.nih.gov/pubmed/33753169 http://dx.doi.org/10.1016/j.jbc.2021.100568 |
work_keys_str_mv | AT rehlingdaniel crystalstructuresofnudt15variantsenabledbyapotentinhibitorrevealthestructuralbasisforthiopurinesensitivity AT zhangsimin crystalstructuresofnudt15variantsenabledbyapotentinhibitorrevealthestructuralbasisforthiopurinesensitivity AT jemthannsofie crystalstructuresofnudt15variantsenabledbyapotentinhibitorrevealthestructuralbasisforthiopurinesensitivity AT koolmeistertobias crystalstructuresofnudt15variantsenabledbyapotentinhibitorrevealthestructuralbasisforthiopurinesensitivity AT throupadam crystalstructuresofnudt15variantsenabledbyapotentinhibitorrevealthestructuralbasisforthiopurinesensitivity AT wallnerolov crystalstructuresofnudt15variantsenabledbyapotentinhibitorrevealthestructuralbasisforthiopurinesensitivity AT scalettiemma crystalstructuresofnudt15variantsenabledbyapotentinhibitorrevealthestructuralbasisforthiopurinesensitivity AT moriyamatakaya crystalstructuresofnudt15variantsenabledbyapotentinhibitorrevealthestructuralbasisforthiopurinesensitivity AT nishiirina crystalstructuresofnudt15variantsenabledbyapotentinhibitorrevealthestructuralbasisforthiopurinesensitivity AT daviesjonathan crystalstructuresofnudt15variantsenabledbyapotentinhibitorrevealthestructuralbasisforthiopurinesensitivity AT desrosesmatthieu crystalstructuresofnudt15variantsenabledbyapotentinhibitorrevealthestructuralbasisforthiopurinesensitivity AT ruddseang crystalstructuresofnudt15variantsenabledbyapotentinhibitorrevealthestructuralbasisforthiopurinesensitivity AT scobiemartin crystalstructuresofnudt15variantsenabledbyapotentinhibitorrevealthestructuralbasisforthiopurinesensitivity AT homanevert crystalstructuresofnudt15variantsenabledbyapotentinhibitorrevealthestructuralbasisforthiopurinesensitivity AT berglundulrikawarpman crystalstructuresofnudt15variantsenabledbyapotentinhibitorrevealthestructuralbasisforthiopurinesensitivity AT yangjunj crystalstructuresofnudt15variantsenabledbyapotentinhibitorrevealthestructuralbasisforthiopurinesensitivity AT helledaythomas crystalstructuresofnudt15variantsenabledbyapotentinhibitorrevealthestructuralbasisforthiopurinesensitivity AT stenmarkpal crystalstructuresofnudt15variantsenabledbyapotentinhibitorrevealthestructuralbasisforthiopurinesensitivity |