Poly(ADP-ribose) polymerase 1 (PARP1) is a promising target for cancer therapy in DNA repair-compromised tumors. One PARP1 inhibitor (Olaparib) is used in the clinic to treat ovarian cancer, and ~20 phase III clinical trials with inhibitors of PARP1 are currently ongoing. PARP1 is a highly abundant nuclear protein that upon binding damaged DNA adds long chains of poly(ADP-ribose) onto itself and many other nuclear proteins. This baroque post-translational modification recruits key components of the DNA repair machinery. Existing inhibitors of PARP1 require a high effective dose (100-400 mg/day) and show poor correlation of drug efficacy with inhibition of PARP1 activity, possibly because they also target inactive PARP1 that is abundantly bound to undamaged chromatin. This PARP1 population has many nuclear functions that extend beyond the maintenance of genome integrity, such as chromatin compaction, and the regulation of transcription and DNA methylation. Our goal is to use rigorous quantitative, structural, and mechanistic approaches to investigate how PARP1 interacts differently with damaged vs. intact DNA in the context of chromatin.
We aim to understand if selective inhibition of the DNA-bound active conformation of PARP1 is a potential approach for increasing drug efficacy and specificity. Using novel enzymatic assays, we will test the varied outcomes of activation by different types of DNA damage. We also propose to investigate the interplay between PARP1 and the much less active and less abundant PARP2. Our studies will reveal the conformational diversity of PARP1 and PARP2, and show how this affects the complex reaction mechanism, resulting in different downstream signaling events. Our mechanistic studies will provide key insights into how to design better screens and assays for the development of more selective, mechanism-based PARP inhibitors. Altogether our proposed research will aid the development of the next generation of PARP inhibitors for use in cancer therapy.

Public Health Relevance

Poly(ADP-ribose) polymerase 1 (PARP1) is a promising target for cancer therapy in DNA repair-compromised tumors with ~20 ongoing phase III clinical trials and one approved drug. We propose to study the structure and activity of PARP1 bound to chromatin DNA. Our quantitative, structural, and mechanistic approaches provide fundamental insight into this abundant multi-faceted enzyme, and will aid in the development of the next generation of PARP inhibitors.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA218255-02
Application #
9513502
Study Section
Macromolecular Structure and Function A Study Section (MSFA)
Program Officer
Witkin, Keren L
Project Start
2017-08-01
Project End
2022-07-31
Budget Start
2018-08-01
Budget End
2019-07-31
Support Year
2
Fiscal Year
2018
Total Cost
Indirect Cost
Name
University of Colorado at Boulder
Department
Chemistry
Type
Graduate Schools
DUNS #
007431505
City
Boulder
State
CO
Country
United States
Zip Code
80303