Chromatin-based DNA damage response (DDR) pathways protect cells from genome and epigenome instability, which are hallmarks of cancer and are thought to drive cancer progression. This proposal addresses questions of fundamental importance for epigenetic and genetic mechanisms involved in cancer. Histone H2A variants are critical components of chromatin but how they promote epigenome and genome stability are poorly understood. For this proposal, we aim to use genetic systems using our histone H2A variant knockout/knockdown engineered human cells with complimentary proteomic analyses of histone variants to answer fundamental questions for how histone H2A variants promote the DDR to maintain genome stability. We also aim to provide insights into how histone H2A variants regulate radiation responses, a therapy often used in cancer treatments. These studies provide innovative and new approaches to these questions that can provide transformative insights into chromatin-based DNA damage responses mediated by histone H2A variants. We will first analyze the DDR function of histone H2A variants in response to radiation, by using human cells deleted or depleted for histone H2A variants including H2AX, H2AZ, macroH2A and H2A.Bbd. Our preliminary results indicate that histone H2A variants play vital and unique roles in these pathways. These studies are poised to provide critical insights into how histone H2A variants regulate the DDR and response to IR. We will then exploit our preliminary analyses identifying candidate protein-interacting factors for each histone H2A variant to identify DDR effector proteins for individual histone H2A variants. Finally, we will test our working hypothesis for how macroH2A promotes DNA repair by HR through the interactions with a new DDR factor that we have identified. These studies will provide an unparalleled view of how histone H2A variants shape the DDR to protect the genome integrity across structurally and functionally diverse chromatin landscapes in human cells. DNA damaging agents are a major class of chemotherapeutic agents including radiotherapy and the epigenome represents exciting new leads for drug discovery. Thus, this work exploits a combination of genetic, biochemical and cellular approaches in human cells to identify key DDR pathways that promote genome and epigenome integrity, as well as responses to cancer-relevant therapies including radiation and PARP inhibitors that can translate to improved treatments for human cancers.

Public Health Relevance

Maintaining genome integrity is fundamental for cellular homeostasis and averting diseases including cancer. Histone H2A variants are key epigenetic components of chromatin that impact mechanisms that govern both genome and epigenome stability. Our goal is to identify the role of histone H2A variants in promoting chromatin-based DNA damage responses that maintain the stability of the genome and epigenome - findings that will significantly impact public health and will provide the molecular insights into DNA-damage and epigenome-based mechanisms involved in the etiology and treatment of human cancers.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA198279-03
Application #
9440374
Study Section
Radiation Therapeutics and Biology Study Section (RTB)
Program Officer
Fingerman, Ian M
Project Start
2016-04-05
Project End
2021-03-31
Budget Start
2018-04-01
Budget End
2019-03-31
Support Year
3
Fiscal Year
2018
Total Cost
Indirect Cost
Name
University of Texas Austin
Department
Biology
Type
Schools of Arts and Sciences
DUNS #
170230239
City
Austin
State
TX
Country
United States
Zip Code
78759
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Zacharioudakis, Emmanouil; Agarwal, Poonam; Bartoli, Alexandra et al. (2017) Chromatin Regulates Genome Targeting with Cisplatin. Angew Chem Int Ed Engl 56:6483-6487
Leung, Justin W C; Makharashvili, Nodar; Agarwal, Poonam et al. (2017) ZMYM3 regulates BRCA1 localization at damaged chromatin to promote DNA repair. Genes Dev 31:260-274
Gong, Fade; Chiu, Li-Ya; Miller, Kyle M (2016) Acetylation Reader Proteins: Linking Acetylation Signaling to Genome Maintenance and Cancer. PLoS Genet 12:e1006272