Cell fate after DNA damage is determined by the cellular DNA damage response (DDR) mechanism that encompasses DNA repair, cell cycle checkpoints, and programmed cell death. Although extensive efforts were dedicated to revealing the activation network of the DDR, little is known about how the cell deactivates the DDR and resumes cell cycle progression, a process termed checkpoint recovery. Importantly, emerging evidence suggests that cancer cells may utilize the recovery mechanism to bypass the DDR during tumorigenesis, and that checkpoint recovery from radio- or chemotherapy causes chemoresistance and tumor recurrence, suggesting checkpoint recovery as an ideal target to improve cancer therapy and patient survival. The long-term goal of this project is to delineate the mechanism of checkpoint recovery and to investigate the implication of checkpoint recovery for cancer progression and therapeutics. We recently characterized the first in vitro system to study checkpoint recovery, and used this system to identify Greatwall (Gwl) kinase as an essential regulator of checkpoint recovery. Interestingly, our preliminary studies discovered novel phosphatases-dependent regulation of checkpoint recovery kinases; our results suggested coordinated activation of Gwl and Plk1, and characterized Gwl phosphorylation and interaction as early events of checkpoint recovery. Moreover, preliminary studies in cancer cell lines and mouse tumor models suggested Gwl as an effective target to enhance chemotherapy and block tumor recurrence. In this proposal we will address fundamental questions regarding the regulatory mechanism of Gwl in checkpoint recovery and its role in cancer therapy and tumor recurrence.
Aim 1 will investigate how Gwl and Plk1 are regulated by DNA damage. Our study will take advantage of the cell-free system and a site-specific phosphatase assay to investigate DNA damage-induced, and phosphatase-dependent regulation of Gwl and Plk1.
Aim 2 will ask how Gwl and Plk1 re-activate from checkpoint arrest. We will characterize novel Gwl/Plk1 interaction and phosphorylation as potential molecular switches that initiate recovery.
In Aim 3, we will characterize how Gwl up-regulation promotes cell recovery from cisplatin treatment and thereby facilitating tumor recurrence. We will then directly evaluate new therapeutic ideas in which targeting Gwl or its initial activation during checkpoint recovery leads to more effective and specific cancer therapy.

Public Health Relevance

Despite that radiation and chemotherapy are major tools for cancer therapeutics, little is understood about how cancer cells escape from these treatment and continue proliferation, a process termed checkpoint recovery. We use comprehensive approaches to delineate the mechanism of this process and directly validate new therapeutic targets for enhanced cancer therapy. PUBLIC HEALTH RELEVANCE: Radiotherapy or chemotherapy kills cancer cells primarily through induction of DNA damage. Thus, mechanisms that allow cells to recover from the DNA damage response and return to cell proliferation are critical for tumor recurrence, and ideal targets for cancer therapy. This project will reveal a new mechanism of DNA damage recovery and tumor recurrence after chemotherapy, and thereby providing better understanding of chemoresistance and potentially leading to more effective cancer therapeutics.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA172574-05
Application #
9250109
Study Section
Basic Mechanisms of Cancer Therapeutics Study Section (BMCT)
Program Officer
Witkin, Keren L
Project Start
2013-04-01
Project End
2018-08-31
Budget Start
2017-04-01
Budget End
2018-08-31
Support Year
5
Fiscal Year
2017
Total Cost
Indirect Cost
Name
University of Nebraska Medical Center
Department
Dentistry
Type
Schools of Dentistry/Oral Hygn
DUNS #
168559177
City
Omaha
State
NE
Country
United States
Zip Code
68198
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Ren, Dapeng; Fisher, Laura A; Zhao, Jing et al. (2017) Cell cycle-dependent regulation of Greatwall kinase by protein phosphatase 1 and regulatory subunit 3B. J Biol Chem 292:10026-10034
Zhu, Songli; Peng, Aimin (2016) Non-homologous end joining repair in Xenopus egg extract. Sci Rep 6:27797
Luong, Khanh V; Wang, Ling; Roberts, Brett J et al. (2016) Cell fate determination in cisplatin resistance and chemosensitization. Oncotarget 7:23383-94
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Fisher, Laura A; Wang, Ling; Wu, Lan et al. (2014) Phosphatase 1 nuclear targeting subunit is an essential regulator of M-phase entry, maintenance, and exit. J Biol Chem 289:23745-52
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