Selective small molecule PARG inhibitor causes replication fork stalling and cancer cell death

Jerry H. Houl; Zu Ye; Chris A. Brosey; Lakshitha P.F. Balapiti-Modarage; Sarita Namjoshi; Albino Bacolla; Daniel Laverty; Brian L. Walker; Yasin Pourfarjam; Leslie S. Warden; Naga Babu Chinnam; Davide Moiani; Roderick A. Stegeman; Mei Kuang Chen; Mien Chie Hung; Zachary D. Nagel; Tom Ellenberger; In Kwon Kim; Darin E. Jones; Zamal Ahmed; John A. Tainer

doi:10.1038/s41467-019-13508-4

Selective small molecule PARG inhibitor causes replication fork stalling and cancer cell death

Jerry H. Houl, Zu Ye, Chris A. Brosey, Lakshitha P.F. Balapiti-Modarage, Sarita Namjoshi, Albino Bacolla, Daniel Laverty, Brian L. Walker, Yasin Pourfarjam, Leslie S. Warden, Naga Babu Chinnam, Davide Moiani, Roderick A. Stegeman, Mei Kuang Chen, Mien Chie Hung, Zachary D. Nagel, Tom Ellenberger, In Kwon Kim, Darin E. Jones, Zamal AhmedJohn A. Tainer

Molecular & Cellular Oncology

Research output: Contribution to journal › Article › peer-review

68 Scopus citations

Abstract

Poly(ADP-ribose)ylation (PARylation) by PAR polymerase 1 (PARP1) and PARylation removal by poly(ADP-ribose) glycohydrolase (PARG) critically regulate DNA damage responses; yet, conflicting reports obscure PARG biology and its impact on cancer cell resistance to PARP1 inhibitors. Here, we found that PARG expression is upregulated in many cancers. We employed chemical library screening to identify and optimize methylxanthine derivatives as selective bioavailable PARG inhibitors. Multiple crystal structures reveal how substituent positions on the methylxanthine core dictate binding modes and inducible-complementarity with a PARG-specific tyrosine clasp and arginine switch, supporting inhibitor specificity and a competitive inhibition mechanism. Cell-based assays show selective PARG inhibition and PARP1 hyperPARylation. Moreover, our PARG inhibitor sensitizes cells to radiation-induced DNA damage, suppresses replication fork progression and impedes cancer cell survival. In PARP inhibitor-resistant A172 glioblastoma cells, our PARG inhibitor shows comparable killing to Nedaplatin, providing further proof-of-concept that selectively inhibiting PARG can impair cancer cell survival.

Original language	English (US)
Article number	5654
Journal	Nature communications
Volume	10
Issue number	1
DOIs	https://doi.org/10.1038/s41467-019-13508-4
State	Published - Dec 1 2019

ASJC Scopus subject areas

General Chemistry
General Biochemistry, Genetics and Molecular Biology
General Physics and Astronomy

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Houl, J. H., Ye, Z., Brosey, C. A., Balapiti-Modarage, L. P. F., Namjoshi, S., Bacolla, A., Laverty, D., Walker, B. L., Pourfarjam, Y., Warden, L. S., Babu Chinnam, N., Moiani, D., Stegeman, R. A., Chen, M. K., Hung, M. C., Nagel, Z. D., Ellenberger, T., Kim, I. K., Jones, D. E., ... Tainer, J. A. (2019). Selective small molecule PARG inhibitor causes replication fork stalling and cancer cell death. Nature communications, 10(1), Article 5654. https://doi.org/10.1038/s41467-019-13508-4

Houl, JH, Ye, Z, Brosey, CA, Balapiti-Modarage, LPF, Namjoshi, S, Bacolla, A, Laverty, D, Walker, BL, Pourfarjam, Y, Warden, LS, Babu Chinnam, N, Moiani, D, Stegeman, RA, Chen, MK, Hung, MC, Nagel, ZD, Ellenberger, T, Kim, IK, Jones, DE, Ahmed, Z & Tainer, JA 2019, 'Selective small molecule PARG inhibitor causes replication fork stalling and cancer cell death', Nature communications, vol. 10, no. 1, 5654. https://doi.org/10.1038/s41467-019-13508-4

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title = "Selective small molecule PARG inhibitor causes replication fork stalling and cancer cell death",

abstract = "Poly(ADP-ribose)ylation (PARylation) by PAR polymerase 1 (PARP1) and PARylation removal by poly(ADP-ribose) glycohydrolase (PARG) critically regulate DNA damage responses; yet, conflicting reports obscure PARG biology and its impact on cancer cell resistance to PARP1 inhibitors. Here, we found that PARG expression is upregulated in many cancers. We employed chemical library screening to identify and optimize methylxanthine derivatives as selective bioavailable PARG inhibitors. Multiple crystal structures reveal how substituent positions on the methylxanthine core dictate binding modes and inducible-complementarity with a PARG-specific tyrosine clasp and arginine switch, supporting inhibitor specificity and a competitive inhibition mechanism. Cell-based assays show selective PARG inhibition and PARP1 hyperPARylation. Moreover, our PARG inhibitor sensitizes cells to radiation-induced DNA damage, suppresses replication fork progression and impedes cancer cell survival. In PARP inhibitor-resistant A172 glioblastoma cells, our PARG inhibitor shows comparable killing to Nedaplatin, providing further proof-of-concept that selectively inhibiting PARG can impair cancer cell survival.",

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AU - Namjoshi, Sarita

AU - Bacolla, Albino

AU - Laverty, Daniel

AU - Walker, Brian L.

AU - Pourfarjam, Yasin

AU - Warden, Leslie S.

AU - Babu Chinnam, Naga

AU - Moiani, Davide

AU - Stegeman, Roderick A.

AU - Chen, Mei Kuang

AU - Hung, Mien Chie

AU - Nagel, Zachary D.

AU - Ellenberger, Tom

AU - Kim, In Kwon

AU - Jones, Darin E.

AU - Ahmed, Zamal

AU - Tainer, John A.

PY - 2019/12/1

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N2 - Poly(ADP-ribose)ylation (PARylation) by PAR polymerase 1 (PARP1) and PARylation removal by poly(ADP-ribose) glycohydrolase (PARG) critically regulate DNA damage responses; yet, conflicting reports obscure PARG biology and its impact on cancer cell resistance to PARP1 inhibitors. Here, we found that PARG expression is upregulated in many cancers. We employed chemical library screening to identify and optimize methylxanthine derivatives as selective bioavailable PARG inhibitors. Multiple crystal structures reveal how substituent positions on the methylxanthine core dictate binding modes and inducible-complementarity with a PARG-specific tyrosine clasp and arginine switch, supporting inhibitor specificity and a competitive inhibition mechanism. Cell-based assays show selective PARG inhibition and PARP1 hyperPARylation. Moreover, our PARG inhibitor sensitizes cells to radiation-induced DNA damage, suppresses replication fork progression and impedes cancer cell survival. In PARP inhibitor-resistant A172 glioblastoma cells, our PARG inhibitor shows comparable killing to Nedaplatin, providing further proof-of-concept that selectively inhibiting PARG can impair cancer cell survival.

AB - Poly(ADP-ribose)ylation (PARylation) by PAR polymerase 1 (PARP1) and PARylation removal by poly(ADP-ribose) glycohydrolase (PARG) critically regulate DNA damage responses; yet, conflicting reports obscure PARG biology and its impact on cancer cell resistance to PARP1 inhibitors. Here, we found that PARG expression is upregulated in many cancers. We employed chemical library screening to identify and optimize methylxanthine derivatives as selective bioavailable PARG inhibitors. Multiple crystal structures reveal how substituent positions on the methylxanthine core dictate binding modes and inducible-complementarity with a PARG-specific tyrosine clasp and arginine switch, supporting inhibitor specificity and a competitive inhibition mechanism. Cell-based assays show selective PARG inhibition and PARP1 hyperPARylation. Moreover, our PARG inhibitor sensitizes cells to radiation-induced DNA damage, suppresses replication fork progression and impedes cancer cell survival. In PARP inhibitor-resistant A172 glioblastoma cells, our PARG inhibitor shows comparable killing to Nedaplatin, providing further proof-of-concept that selectively inhibiting PARG can impair cancer cell survival.

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Selective small molecule PARG inhibitor causes replication fork stalling and cancer cell death

Abstract

ASJC Scopus subject areas

MD Anderson CCSG core facilities

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