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Targeted Protein Degradation of NUDT5 Dissociates Catalytic Inhibition from Protein Loss in 6-Thioguanine Response

Anne-Sophie M C Marques1,2, Ludwig G Bauer1,2, Tuan-Anh Nguyen3

  • 1Centre for Medicines Discovery, Nuffield Department of Medicine, University of Oxford, Old Road Campus, Roosevelt Drive, Oxford, UK.

Nature Communications
|June 30, 2026
PubMed

Insights

6-Thioguanine (6-TG) drug toxicity is mediated by NUDT5, not just its enzymatic activity. Developing NUDT5 degraders reveals a non-enzymatic role, offering new therapeutic strategies for leukemia treatment.

Area of Science:

  • Biochemistry
  • Pharmacology
  • Genetics

Background:

  • 6-Thioguanine (6-TG) is an FDA-approved antimetabolite drug used clinically for leukemia treatment.
  • Cellular effects of 6-TG depend on metabolic activation and protein interactions, notably with NUDT15, which hydrolyzes active 6-TG metabolites.
  • NUDT5, another NUDIX hydrolase, has been identified as a key mediator of 6-TG toxicity.

Purpose of the Study:

  • To develop and validate a selective, cell-active NUDT5 degrader toolkit.
  • To characterize target engagement, degradation kinetics, and proteome-wide selectivity of NUDT5 degraders.
  • To elucidate the role of NUDT5 in modulating cellular response to 6-TG.

Main Methods:

  • Development and validation of selective NUDT5 degraders.
  • Orthogonal characterization of target engagement and degradation kinetics.
  • CRISPR loss-of-function studies and reconstitution experiments.
  • Proteome-wide selectivity profiling.

Main Results:

  • NUDT5 protein depletion antagonizes NUDT15 inhibition.
  • Degraders and CRISPR experiments support a non-enzymatic role for NUDT5 in 6-TG response.
  • Selective NUDT5 degraders were successfully developed and validated.

Conclusions:

  • NUDT5 plays a crucial, non-enzymatic role in 6-thioguanine toxicity.
  • NUDT5's distinct mode-of-action from NUDT15 has potential therapeutic implications for leukemia patients.
  • Targeting NUDT5 offers a novel therapeutic strategy beyond NUDT15 inhibition.

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