PARP1 inhibitors (PARPi) are known to kill tumor cells via two mechanisms (PARP1 catalytic inhibition and PARP1 trapping). The relative contribution of these two pathways in mediating the cytotoxicity of PARPi, however, is not well understood. Here we designed a series of small molecule PARP degraders. Treatment with one such compound iRucaparib-AP6 results in highly efficient and specific PARP1 degradation. iRucaparib-AP6 blocks the enzymatic activity of PARP1 in vitro, and PARP1-mediated poly-ADP-ribosylation signaling in intact cells. This strategy mimics PARP1 genetic depletion, which enables the pharmacological decoupling of PARP1 inhibition from PARP1 trapping. Finally, by depleting PARP1, iRucaparib-AP6 protects muscle cells and primary cardiomyocytes from DNA-damage-induced energy crisis and cell death. In summary, these compounds represent ‘non-trapping’ PARP1 degraders that block both the catalytic activity and scaffolding effects of PARP1, providing an ideal approach for the amelioration of the various pathological conditions caused by PARP1 hyperactivation.
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The MS data have been deposited in the ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifiers PXD014838 (Supplementary Dataset 1), PXD014836 and PXD014837 (Supplementary Datasets 2 and 3), PXD014840 (Supplementary Datasets 4 and 5), and PXD014839 (Supplementary Dataset 6). Source data for Figs. 2d, 5d and 6e,f, and Supplementary Fig. 12a–f are available online. Full uncropped blots are shown in Supplementary Fig. 14.
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We thank J. A. Hill (UT Southwestern Medical Center) for sharing primary rat neonatal cardiomyocytes and X. Zhong and C. Kim for help with the immunofluorescence microscopy experiments and PARP1 immunoprecipitation experiments, respectively. We thank X. Yu (City of Hope) for providing the GFP-PARP1 construct. We also thank X. D. Wang, L. Yuan and the other members of the Yu laboratory for helpful discussions. This work was supported by grants from the National Institutes of Health (GM122932 to Y.Y. and CA226419 to C.C.) and the Welch foundation (I-1800 to Y.Y.).
Y.Y. receives research support from Pfizer. A provisional patent application on the PARP degraders and technologies described herein has been filed by Y.Y., C.C., S. W. and L. H.
Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Figures 1–14 and Supplementary Table 1
Quantified proteomic data for primary cardiomyocytes treated with DMSO, iRucaparib-AP5 or iRucaparib-AP6 (1 μM) for 24 h.
Quantified proteomic data for HeLa cells treated with DMSO, iRucaparib-TP3 (5 μM) or iRucaparib-TP3 (5 μM) plus Rucaparib (1 μM) for 24 h.
Quantified proteomic data for BT-549 cells treated with DMSO, iRucaparib-TP3 (5 μM) or vRucaparib-TP4 (20 μM) for 24 h.
Quantified PARylated proteomic data for SILAC-labeled HeLa cells treated with DMSO and Rucaparib or DMSO and iRucaparib-AP6 (10 μM).
Quantified PARylated proteomic data for SILAC-labeled HeLa cells treated with DMSO and Rucaparib or DMSO and iRucaparib-TP3 (10 μM).
Quantified chromatin proteomic data for HeLa cells treated with MMS and DMSO, MMS and Rucaparib or MMS and iRucaparib-AP6 for 24 h.
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Wang, S., Han, L., Han, J. et al. Uncoupling of PARP1 trapping and inhibition using selective PARP1 degradation. Nat Chem Biol 15, 1223–1231 (2019) doi:10.1038/s41589-019-0379-2