Defining a protein hydroxylase tumour suppressor pathway

Lead Research Organisation: University of Birmingham
Department Name: Cancer Sciences

Abstract

Normal cell growth and survival is tightly controlled by multiple overlapping biochemical pathways that work together to ensure that cells only divide at the appropriate time and only survive in the correct environment. When these fail-safe mechanisms go wrong the uncontrolled cell division that results can initiate the early stages of cancer development. These so called 'tumour suppressor' pathways are often inactivated in cancer due to mutations in the DNA of specific genes.

We have identified a gene that when experimentally mutated promotes tumourigenesis in cell models. Consistent with a role in human cancer this gene is also mutated in human tumour samples. This new tumour suppressor gene, called 'MINA', encodes a protein with enzyme activity. Through this activity MINA modifies other proteins with oxygen in a process called 'hydroxylation'. We propose that MINA normally helps promote healthy cell division and survival by hydroxylating proteins that control the expression, or 'transcription', of key cancer-associated genes.

In this proposal we aim to characterise the role and regulation of MINA in cancer. We will first investigate whether MINA is inactivated by mechanisms other than gene mutation. Specifically, we will ask whether the MINA protein is itself modified, and how this modification switches off its enzyme activity in cancer cells. Then we will use a combination of biochemical and genomic approaches to identify the pathways by which MINA activity suppresses cancer. We will use 'mass spectrometry', a biophysical technique that measure the mass of protein fragments, to identify new proteins that MINA hydroxylates, and a DNA sequencing technique called 'ChIP-seq' to identify gene transcription regulated by MINA. Finally, we will validate our findings by measuring our biomarkers of MINA regulation and targets in several hundred human tumour samples.

Through these approaches we hope to define a novel tumour suppressor pathway involving a hydroxylase enzyme. Because this class of enzyme is not widely appreciated to play an important role in cancer, the work will stimulate further research in this important area. Furthermore, by discovering the key players in this new tumour suppressor pathway the work may identify new therapeutic opportunities for cancer treatments. Importantly, because MINA has important but poorly characterised roles in other disease-associated pathways, particularly in the immune system, the findings will also be of interest to a wide variety of other medical researchers.

Technical Summary

Deregulated protein modifications have been widely implicated in the pathogenesis of major human diseases. The discovery and characterisation of novel post-translational modifications (PTM) has opened new avenues of biomedical research and stimulated new therapeutic opportunities.

Hydroxylation is an under-studied but emerging PTM. Pioneering work on collagen and oxygen sensing highlighted the potential importance of hydroxylation in physiology and pathology, and related oxygenases have since been implicated in a variety of important disease-associated processes. However, several members of the hydroxylase family are poorly characterised and the extent and functions of protein hydroxylation in disease remain unclear.

We previously reported that a nuclear JmjC protein called 'Myc-Induced Nuclear Antigen' (MINA) is a histidine hydroxylase. Our functional studies indicate that MINA has tumour suppressor (TS) activity, consistent with the observation that it is mutated and inactivated in a variety of tumour types, including gastrointestinal cancers. However, a detailed understanding of the extent of MINA deregulation in cancer requires knowledge of other potential mechanisms of inactivation. Furthermore, the downstream consequences of MINA inhibition are not known. Here we propose to address these limitations by characterising the regulation of MINA by an oncogenic kinase and identifying novel cancer-relevant substrates and gene targets involved in MINA-dependent transcriptional control. Importantly, the findings will be extended into an independent cohort of several hundred gastric tumour samples in a combined approach involving targeted resequencing, gene expression analyses and histopathology.

The work will improve our understanding of protein hydroxylation and its role in fundamental cellular processes, define a novel tumour suppressor pathway, and highlight new therapeutic opportunities for the treatment of cancer and other diseases.

Planned Impact

The work proposed here aims to significantly advance our understanding of a poorly characterised protein modification (hydroxylation) by defining its role in regulating transcriptional control in a novel tumour suppressor pathway. The work will deliver significant scientific advances that will benefit academic and industrial researchers working on cancer and other important diseases. Therefore, the project has the potential to deliver impact by enhancing health and in the future to lead to the development of novel therapeutic opportunities.

The areas in which this proposal will provide meaningful impact are:

1) Academic
The work will engage a number of international academic beneficiaries, including; fundamental cell biology researchers, researchers and clinicians with an interest in mechanisms of Aurora Kinase A-mediated tumourigenesis and gastrointestinal cancer, and immunologists investigating the role of pro-inflammatory T-cells in autoimmune disease (including our collaborator, Mark Bix).

2) Commercial
The identification of novel chemotherapeutic targets has classically followed the discovery and characterisation of oncogene signalling pathways and tumour suppressor genes. The work proposed here will be important to the pharmaceutical industry for two reasons. Firstly, it will highlight the risk of small molecule inhibition of the MINA hydroxylase in cancer (which is being promoted by some academic groups). Secondly, it will define a novel tumour suppressor pathway that will help identify new therapeutic opportunities. By delivering a very high level of scientific proof the work will provide a strong foundation for successful drug discovery.

Beyond cancer MINA is attracting interest as a therapeutic target in autoimmune disease due to an important role in transcriptional regulation of the TH17 differentiation program. Through the proposed work identifying the transcriptional targets of MINA the findings will be of importance to the commercial sector in this area.

3) Socio-economic
Cancer is a major worldwide societal and economic burden that is increasing due to a growing and ageing population. Several major cancers are still associated with poor survival rates, due to a variety of factors including poor treatment options, resistance to targeted therapies and heterogeneity complicating molecular classification. The work proposed here uses gastrointestinal cancers as a model to define a novel tumour suppressor pathway. Therefore, the findings will likely benefit patients in the future and also potentially reduce public costs and improve the quality of health care.

4) Training and education
This proposal involves combined biochemical, cell biology, genomic and pathology approaches that will provide exceptional training opportunities for the next generation of young scientists. This applies not only to the staff directly involved in the project, but also interns and undergraduate, MSc and PhD students that the Coleman group regularly host.

Publications

10 25 50
 
Description Cancer Research UK PhD Studentship
Amount £140,000 (GBP)
Funding ID 25674 
Organisation Cancer Research UK 
Sector Charity/Non Profit
Country United Kingdom
Start 10/2018 
End 10/2022
 
Description Cancer Research UK Program Foundation Award
Amount £1,200,000 (GBP)
Funding ID 24552 
Organisation Cancer Research UK 
Sector Charity/Non Profit
Country United Kingdom
Start 11/2017 
End 10/2023
 
Description Research Project Grant - MCMB Panel
Amount £400,000 (GBP)
Funding ID MR/N021053/1 
Organisation Medical Research Council (MRC) 
Sector Public
Country United Kingdom
Start 07/2016 
End 06/2019
 
Description Bioinformatic analyses of oxygenase mutation in cancer 
Organisation University of Birmingham
Department Centre for Computational Biology
Country United Kingdom 
Sector Academic/University 
PI Contribution We have provided candidate oxygenases for assessment of mutation status in cancer NGS studies
Collaborator Contribution Our collaborator, Dr Andrew Beggs, has provided detailed information regarding the mutation status of several oxygenases across cancer.
Impact In progress.
Start Year 2014
 
Description Genomic characterisation of a histidine hydroxylase 
Organisation Chiba University
Country Japan 
Sector Academic/University 
PI Contribution Cell models, reagents and expertise for studying the genomic localisation of a histidine hydroxylase by ChIP-seq
Collaborator Contribution Expertise, reagents, protocols and guidance for studying the genomic localisation of a histidine hydroxylase by ChIP-seq
Impact This project is multidisciplinary as it crosses our expertise in tumour cell biology and the expertise of our collaborator in immunology.
Start Year 2016
 
Description Histology analyses of protein hydroxylases in gastric cancer 
Organisation University of Birmingham
Country United Kingdom 
Sector Academic/University 
PI Contribution Intellectual input related to project development and identification of relevant human tumour samples
Collaborator Contribution Storage, retrieval, processing and histology analyses of human tumour samples
Impact not as yet.
Start Year 2016
 
Description Investigating the regulation of protein hydroxyalses by mitotic kinases 
Organisation University of Rennes 1
Country France 
Sector Academic/University 
PI Contribution Cell models and reagents for investigating the regulation of protein hydroxyalses by mitotic kinases
Collaborator Contribution Reagents, protocols, guidance and advice for investigating the regulation of protein hydroxyalses by mitotic kinases
Impact None as yet.
Start Year 2016
 
Description Mass spectrometry analyses of hydroxylase targets 
Organisation University of Oxford
Country United Kingdom 
Sector Academic/University 
PI Contribution Provision of protein samples for MS analyses.
Collaborator Contribution MS expertise
Impact In progress.
 
Description Pathology analyses of protein hydroxylases in cancer 
Organisation University of Birmingham
Department Molecular Pathology Centre
Country United Kingdom 
Sector Academic/University 
PI Contribution Intellectual input and project development related to the identification and retrieval of appropriate tumour samples and biomarkers for pathology analyses.
Collaborator Contribution Pathology analyses of appropriate tumour samples following immunohistochemical staining of specific biomarkers.
Impact not as yet.
Start Year 2018
 
Description Fundraising Dinner for CRUK 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Industry/Business
Results and Impact A fundraising event organised by theBirmingham Business Committee - a small group of volunteers made up of senior managers from Birmingham's business community. 200 guests in attendance, with proceeds supporting CRUK research happening at the University of Birmingham.
One researcher sat on each table and informally talked to the guests during the evening about cancer research (eg what you are working on, what difference this could ultimately make to people with cancer, particular research strengths in Birmingham etc), as well as generally network during the drinks reception and meal. Mayor Andy Street attended.
Year(s) Of Engagement Activity 2018
 
Description Lab Tour 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Supporters
Results and Impact Visit by a group of our CRUK Race for Life volunteers and Events staff.
Year(s) Of Engagement Activity 2017
 
Description Lab Tour 
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 Industry/Business
Results and Impact large visit/lab tour for one of CRUK's major corporate partner (this company has raised over £4 million for CRUK).Feedback from DEbbie Ringham CRUK public engagement office:"A huge thanks for all of your support with the Beiersdorf leadership team visit yesterday. This is a major corporate partner of Cancer Research UK, who has raised around £4 million to date for our work. It was brilliant for me to be able to offer their entire team the chance to come and learn more about what we do at their nearest CRUK Centre, and I really appreciate you taking the time to support this visit"
Year(s) Of Engagement Activity 2017
 
Description Lab Tour 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Supporters
Results and Impact 2 charity ball organisers visited the lab. Here is the feedback:"I just wanted to say many thanks for inviting myself and Jen to the lab tours. We found it incredibly interesting and it was great to meet some of the phd students who are clearly very talented.
Jen and I are thinking of organising another ball for October 2018 and will be in touch later on in the year to have a chat with you about it"
And this from CRUK engagement office Debbie Ringham "Thank you both - and fantastic that they have been inspired from this to put on another fundraiser for us "
Year(s) Of Engagement Activity 2017
 
Description Lab Tour 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Industry/Business
Results and Impact Evening lab tour for a local Women in Business Association
Year(s) Of Engagement Activity 2018
 
Description Lab Tour 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Industry/Business
Results and Impact Visit by Shakespeare Martineau, a Birmingham based legal firm who are fundraising for CRUK.
Year(s) Of Engagement Activity 2018
 
Description Lab Tour 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Supporters
Results and Impact Lab Tour for CRUK Legacy team and potential donors.
Year(s) Of Engagement Activity 2018
 
Description Lab tour 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Supporters
Results and Impact Lab tour for CRUK trading staff and volunteers. I also gave a 20min presentation titled 'What is Cancer?' and and overview of our labs work.
Year(s) Of Engagement Activity 2018
 
Description Open day for CRUK fundraisers 
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 Supporters
Results and Impact Presented sessions in the laboratories on tumour cell culture and microscopy to three groups of CRUK fundraisers
Year(s) Of Engagement Activity 2016
 
Description Public Speech 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Supporters
Results and Impact Gave a talk about the importance of charitable donations for funding cancer research, and an overview of our labs work, at the 'Vaughn Singers' concert
Year(s) Of Engagement Activity 2018