Valerie Nicholas C K, Sanjiv Kumar, Mortusewicz Oliver, Zhang Si Min, Alam Seher, Pires Maria J, Stigsdotter Hannah, Rasti Azita, Langelier Marie-France, Rehling Daniel, Throup Adam, Purewal-Sidhu Oryn, Desroses Matthieu, Onireti Jacob, Wakchaure Prasad, Almlöf Ingrid, Boström Johan, Bevc Luka, Benzi Giorgia, Stenmark Pål, Pascal John M, Helleday Thomas, Page Brent D G, Altun Mikael
Science for Life Laboratory, Division of Clinical Physiology, Department of Laboratory Medicine, Karolinska Institutet, Karolinska University Hospital, Huddinge, SE-141 52, Sweden.
Science for Life Laboratory, Department of Oncology-Pathology, Karolinska Institutet, Solna, SE-171 65, Sweden.
Nat Commun. 2024 Dec 6;15(1):10347. doi: 10.1038/s41467-024-54415-7.
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.
细胞靶点结合技术能够对细胞内药物结合进行定量分析；然而，同时评估与药物相关的表型却颇具挑战性。在此，我们提出了一种基于突变体积累的细胞靶点结合方法，作为一个平台，该平台可以使用条件稳定的药物生物传感器同时评估药物-靶点相互作用和表型反应。我们观察到，在已知药物靶点的报道突变体中，药物反应性蛋白型普遍存在。兼容的突变体似乎遵循结构和生物物理逻辑，这允许生物传感器库在蛋白质内部和旁系同源物中进行扩展。然后，我们应用我们的方法来区分MutT同源物1（MTH1）抑制剂不同细胞活性中的靶点结合情况，剖析具有R139C药物遗传变异的Nudix水解酶15（NUDT15）相关的硫嘌呤代谢，并描绘聚（ADP-核糖）聚合酶1/2（PARP1/2）的结合动力学以及PARP抑制剂（PARPi）对DNA的捕获情况。此外，PARP1衍生的生物传感器有助于对PARP1结合剂进行高通量筛选，以及对活体动物中PARPi结合进行多模态离体分析和非侵入性追踪。这种方法可以通过连接药物结合事件及其生物学后果，促进对药物-靶点结合的全面评估。
