Coupling cellular drug-target engagement to downstream pharmacology with CeTEAM
Cellular target engagement technologies enable quantification of intracellular drug binding; however, simultaneous assessment of drug-associated phenotypes has proven challenging. Here, we present cellular target engagement by accumulation of mutant as a platform that can concomitantly evaluate drug-target interactions and phenotypic responses using conditionally stabilized drug biosensors. We observe that drug-responsive proteotypes are prevalent among reported mutants of known drug targets. Compatible mutants appear to follow structural and biophysical logic that permits intra-protein and paralogous expansion of the biosensor pool. We then apply our method to uncouple target engagement from divergent cellular activities of MutT homolog 1 (MTH1) inhibitors, dissect Nudix hydrolase 15 (NUDT15)-associated thiopurine metabolism with the R139C pharmacogenetic variant, and profile the dynamics of poly(ADP-ribose) polymerase 1/2 (PARP1/2) binding and DNA trapping by PARP inhibitors (PARPi). Further, PARP1-derived biosensors facilitated high-throughput screening for PARP1 binders, as well as multimodal ex vivo analysis and non-invasive tracking of PARPi binding in live animals. This approach can facilitate holistic assessment of drug-target engagement by bridging drug binding events and their biological consequences.
Authors
- Mikael Altun (ORCID: https://orcid.org/0000-0002-6937-6124)
- Kumar Sanjiv (ORCID: https://orcid.org/0000-0002-5415-9952)
- John M. Pascal (ORCID: https://orcid.org/0000-0002-2714-4317)
- Seher Alam (ORCID: https://orcid.org/0000-0002-8494-832X)
- Maria João Pires (ORCID: https://orcid.org/0000-0003-1036-3326)
- Thomas Helleday (ORCID: https://orcid.org/0000-0002-7384-092X)
- Giorgia Benzi
- Prasad B. Wakchaure (ORCID: https://orcid.org/0000-0003-2828-4642)
- Hannah Stigsdotter
- Pål Stenmark (ORCID: https://orcid.org/0000-0003-4777-3417)
- Nicholas C.K. Valerie (ORCID: https://orcid.org/0000-0002-9423-964X)
- Johan Boström (ORCID: https://orcid.org/0000-0001-5252-4023)
- Si Min Zhang (ORCID: https://orcid.org/0000-0001-7763-603X)
- Matthieu Desroses (ORCID: https://orcid.org/0000-0003-4152-3855)
- Adam Throup (ORCID: https://orcid.org/0000-0001-6512-8679)
- Jacob Onireti
- Luka Bevc
- Oliver Mortusewicz (ORCID: https://orcid.org/0000-0002-4290-4994)
- Daniel Rehling (ORCID: https://orcid.org/0000-0002-8627-3469)
- Ingrid Almlöf
- Azita Rasti
- Brent D. G. Page (ORCID: https://orcid.org/0000-0002-2101-1329)
- Marie-France Langelier (ORCID: https://orcid.org/0000-0001-9815-1840)
- Oryn Purewal-Sidhu (ORCID: https://orcid.org/0000-0003-1567-9389)
Institutions
- Karolinska University Hospital (SE)
- University of British Columbia (CA)
- Stockholm University (SE)
- Science for Life Laboratory (SE)
- Karolinska Institutet (SE)
- Université de Montréal (CA)
Publication Details
- Journal
- Nature Communications
- Published
- 2024-12-06
- DOI
- https://doi.org/10.1038/s41467-024-54415-7
- Citations
- 6
- Primary Topic
- PARP inhibition in cancer therapy
- Type
- article
- Field-Weighted Citation Impact
- 1.70
Funders
- Torsten Söderbergs Stiftelse
- Loo och Hans Ostermans Stiftelse för Medicinsk Forskning
- Pain Relief Foundation
- Michael Smith Health Research BC
- European Commission
- Cancerfonden
- Svenska Sällskapet för Medicinsk Forskning
- Karolinska Institutet
- Novo Nordisk
- Alfred Österlunds Stiftelse
- Barncancerfonden
- Science for Life Laboratory
- Novo Nordisk Fonden
- Stiftelsen Felix Mindus Bidrag till Leukemiforskningen
- Horizon 2020 Framework Programme
- Canadian Institutes of Health Research