Defining the role of ADP-ribosyltransferases in DNA repair and genome stability
Lead Research Organisation:
University of Oxford
Department Name: Biochemistry
Abstract
Preserving the integrity of genetic material through repair of damaged DNA is critical for the health of an organism. My laboratory is focussed on understanding these processes, with specific reference to how a set of enzymes called ADP-ribosyltransferases (ARTs) regulate a variety of DNA repair processes.
The best defined role of ARTs is in promoting repair of breaks in the DNA double strand helix. Inhibition of this pathway using small molecule ART inhibitors (ARTi) is being exploited in the clinic to specifically kill tumours with defects in a DNA repair process known as homologous recombination. However, there are several different types of ART in a cell and an emerging theme is that these enzymes can regulate different DNA repair processes. The proposed work will build on our recent exciting new findings addressing how different ARTs function together to repair DNA damage and how inhibiting one ART over others more effectively kills cells following exposure to agents that induce DNA damage. Given currently available ARTi target multiple ARTs, these studies will provide information that underpins the development of more specific ARTi with increased efficacy in the clinic. We will also define novel factors that when inhibited along with ARTs either exaggerate or rescue the sensitivity of cells to DNA damage, identifying pathways that make ART inhibited cells more sensitive or tolerant to radio- or chemotherapy. This work will lead to efforts in improving the efficacy of these agents in the clinic and identify new targets whose inhibition will overcome drug resistance.
The best defined role of ARTs is in promoting repair of breaks in the DNA double strand helix. Inhibition of this pathway using small molecule ART inhibitors (ARTi) is being exploited in the clinic to specifically kill tumours with defects in a DNA repair process known as homologous recombination. However, there are several different types of ART in a cell and an emerging theme is that these enzymes can regulate different DNA repair processes. The proposed work will build on our recent exciting new findings addressing how different ARTs function together to repair DNA damage and how inhibiting one ART over others more effectively kills cells following exposure to agents that induce DNA damage. Given currently available ARTi target multiple ARTs, these studies will provide information that underpins the development of more specific ARTi with increased efficacy in the clinic. We will also define novel factors that when inhibited along with ARTs either exaggerate or rescue the sensitivity of cells to DNA damage, identifying pathways that make ART inhibited cells more sensitive or tolerant to radio- or chemotherapy. This work will lead to efforts in improving the efficacy of these agents in the clinic and identify new targets whose inhibition will overcome drug resistance.
Technical Summary
This work is concerned with increasing our understanding of how different ADP-ribosyltransferases (ARTs) maintain genome integrity through DNA repair. We have exploited genome editing technology to generate cell lines defective in the principle DNA damage responsive ARTs alone, or in combination. Using this unique set of reagents we have identified exciting and novel redundancy between ARTs in resolving DNA damage during S-phase. We will build on this work using genome editing technology, in combination with cutting edge biochemical and cell based assays, to define how the ARTs PARP1 and PARP2 regulate these processes. These approaches will also be extended to characterising a novel ART we have uncovered that allows cells deficient in PARP1 and PARP2 to tolerate DNA damage. Using state of the art screening facilities in Oxford, we will perform a genome-wide CRISPR/Cas9 based screen to identify novel genes and pathways that when disrupted suppress or enhance the sensitivity of PARP-deficient cells to DNA damage. Given inhibition of ARTs is being exploited to treat homologous recombination-defective tumours, we will assess how disruption of these genes impacts on these events. These experiments will lead to efforts in improving the efficacy of these agents in the clinic and identify new targets whose inhibition will overcome drug resistance.
Planned Impact
This work fits within the MRC discovery research priority of 'Resilience, Repair and Replacement', specifically to the 'Natural Protection' objective, and has the potential to impact on human health and well-being in the short and longer terms.
This work constitutes Discovery Research. As such, it will have some immediate and many potential long-term impacts. Primary immediate beneficiaries will be the scientific community, as outlined in the 'Academic Beneficiaries' section. Longer term impacts are rooted in our increased understanding of fundamental concepts in DNA metabolism and how this impacts on human health and clinical practice. Synthetic lethality is emerging as an effective treatment for subsets of tumours defective in components of the DNA damage response. Therefore, in the longer term, this work will increase our understanding of this concept and provide important information that can be exploited to further refine these treatments. Additionally, they will identify potential biomarkers for malignancy, or resistance of tumours to chemo and/or radiotherapy. Longer term beneficiaries will include:
i) Commercial exploitation
- This work will provide commercial companies with information to develop therapies that specifically target malignant cells either alone or in combination with chemo/radiotherapy.
- It will identify genes that when deregulated render cells refractory to treatment with ARTi and/or DNA damaging agents. This will provide companies with potential therapeutic targets that when inhibited will overcome this resistance.
- It will provide information for companies wishing to screen for gene mutations in DNA repair genes that contribute towards malignancy, or resistance of tumours to clinical intervention. This may be extended to a variety of other disease states associated with defects in the DNA damage response including premature ageing, immune deficiencies and neurological degeneration.
ii) Public sector exploitation
- This work will provide information to increase efficacy of cancer treatment
- It will influence policy decisions regarding targeting the DNA damage response in the clinic both in terms of application to drug resistance mechanisms and how this may be overcome, in addition to applying this knowledge to disease states other than cancer.
iii) Wider public in general
- Developing new targeted cancer therapies will impact on the lives of future cancer patients and their families.
- An important impact is to inform the wider public of the importance of genome maintenance in human health and well-being. This will inform tailored, specific information regarding life-style, diet etc. to protect from the effects of DNA damage which accumulate during the aging process. This will allow lifestyles changes that will contribute to health and wellbeing.
This work constitutes Discovery Research. As such, it will have some immediate and many potential long-term impacts. Primary immediate beneficiaries will be the scientific community, as outlined in the 'Academic Beneficiaries' section. Longer term impacts are rooted in our increased understanding of fundamental concepts in DNA metabolism and how this impacts on human health and clinical practice. Synthetic lethality is emerging as an effective treatment for subsets of tumours defective in components of the DNA damage response. Therefore, in the longer term, this work will increase our understanding of this concept and provide important information that can be exploited to further refine these treatments. Additionally, they will identify potential biomarkers for malignancy, or resistance of tumours to chemo and/or radiotherapy. Longer term beneficiaries will include:
i) Commercial exploitation
- This work will provide commercial companies with information to develop therapies that specifically target malignant cells either alone or in combination with chemo/radiotherapy.
- It will identify genes that when deregulated render cells refractory to treatment with ARTi and/or DNA damaging agents. This will provide companies with potential therapeutic targets that when inhibited will overcome this resistance.
- It will provide information for companies wishing to screen for gene mutations in DNA repair genes that contribute towards malignancy, or resistance of tumours to clinical intervention. This may be extended to a variety of other disease states associated with defects in the DNA damage response including premature ageing, immune deficiencies and neurological degeneration.
ii) Public sector exploitation
- This work will provide information to increase efficacy of cancer treatment
- It will influence policy decisions regarding targeting the DNA damage response in the clinic both in terms of application to drug resistance mechanisms and how this may be overcome, in addition to applying this knowledge to disease states other than cancer.
iii) Wider public in general
- Developing new targeted cancer therapies will impact on the lives of future cancer patients and their families.
- An important impact is to inform the wider public of the importance of genome maintenance in human health and well-being. This will inform tailored, specific information regarding life-style, diet etc. to protect from the effects of DNA damage which accumulate during the aging process. This will allow lifestyles changes that will contribute to health and wellbeing.
Publications
Pears CJ
(2021)
Dictyostelium discoideum as a Model to Assess Genome Stability Through DNA Repair.
in Frontiers in cell and developmental biology
Richards F
(2023)
Regulation of Rad52-dependent replication fork recovery through serine ADP-ribosylation of PolD3.
in Nature communications
Ronson GE
(2018)
PARP1 and PARP2 stabilise replication forks at base excision repair intermediates through Fbh1-dependent Rad51 regulation.
in Nature communications
Sharma AB
(2023)
C16orf72/HAPSTR1/TAPR1 functions with BRCA1/Senataxin to modulate replication-associated R-loops and confer resistance to PARP disruption.
in Nature communications
Description | Identification of ADP-ribosylation sites induced upon replication stress |
Organisation | University of Copenhagen |
Country | Denmark |
Sector | Academic/University |
PI Contribution | We provided information and samples for the analysis |
Collaborator Contribution | Mapping ADP-ribosylation sites in our samples using mass spectrometry. |
Impact | Currently writing a manuscript. |
Start Year | 2020 |
Description | Cold Spring Harbor Conference - The PARP family and ADP-ribosylation |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | International Confernece presentation at Cold Spring Harbor - The PARP family and ADP-ribosylation |
Year(s) Of Engagement Activity | 2020 |
Description | Conference Talk: Academic Forum of Academic Strategic Alliance: Oxford and NCKU, Taiwan |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | Research Talk to promote collaborations between Oxford and NCKU, Taiwan. |
Year(s) Of Engagement Activity | 2019 |
Description | Conference talk - PARP2021, Barcelona |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | Conference presentation of research. |
Year(s) Of Engagement Activity | 2021 |
Description | Conference talk - Responses to DNA damage, Netherlands |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | Conference presentation of research |
Year(s) Of Engagement Activity | 2022 |
Description | DNA damage response in cell physiology and disease |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | Scientific conference |
Year(s) Of Engagement Activity | 2022 |
Description | Gordon Research Conference - Mutagenesis |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | Oral communication at conference |
Year(s) Of Engagement Activity | 2018 |
Description | Invited Seminar - AstraZeneca |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Industry/Business |
Results and Impact | Talk to industrial collaborators |
Year(s) Of Engagement Activity | 2018 |
Description | Open Day Department of Biochemistry, University of Oxford |
Form Of Engagement Activity | Participation in an open day or visit at my research institution |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Schools |
Results and Impact | Hosting Open Day at the Department of Biochemistry, September 2017 |
Year(s) Of Engagement Activity | 2017 |
Description | Open Day, Department of Biochemistry, University of Oxford |
Form Of Engagement Activity | Participation in an open day or visit at my research institution |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Schools |
Results and Impact | Hosting Open Day at the Department of Biochemistry, June 2018 |
Year(s) Of Engagement Activity | 2018 |
Description | Open Day, Department of Biochemistry, University of Oxford |
Form Of Engagement Activity | Participation in an open day or visit at my research institution |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Schools |
Results and Impact | Hosting Open Day at the Department of Biochemistry, September 2018 |
Year(s) Of Engagement Activity | 2018 |
Description | Open Day, Dept Biochemistry, University of Oxford |
Form Of Engagement Activity | Participation in an open day or visit at my research institution |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Public/other audiences |
Results and Impact | Departmental Open day for prospective undergraduates |
Year(s) Of Engagement Activity | 2019 |
Description | Open Day, Dept Biochemistry, University of Oxford |
Form Of Engagement Activity | Participation in an open day or visit at my research institution |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Schools |
Results and Impact | Open day for prospective undergraduates |
Year(s) Of Engagement Activity | 2020 |
Description | Open day, Department of biochemistry, university of Oxford |
Form Of Engagement Activity | Participation in an open day or visit at my research institution |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Schools |
Results and Impact | September open day to publicise studying biochemistry at the university of Oxford. |
Year(s) Of Engagement Activity | 2017 |
Description | Research Seminar - University of Sussex |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Professional Practitioners |
Results and Impact | Research Seminar - Genome Damage and Stability Center, University of Sussex |
Year(s) Of Engagement Activity | 2019 |
Description | Schools Open day, Department of Biochemistry, university of Oxford |
Form Of Engagement Activity | Participation in an open day or visit at my research institution |
Part Of Official Scheme? | No |
Geographic Reach | National |
Primary Audience | Schools |
Results and Impact | June Open day for schools to publicise studying biochemistry at Oxford |
Year(s) Of Engagement Activity | 2017 |