The role of microRNAs in paediatric malignant germ cell tumours (MGCTs) and germ cell development
Lead Research Organisation:
MRC Cancer Unit
Department Name: UNLISTED
Abstract
Aims: To investigate the role of tiny genetic regulators (microRNAs) in childhood malignant germ cell tumours (MGCTs) to improve management of such cancers.
Research Proposal: MGCTs come from egg- or sperm-producing cells, representing 4% of childhood cancers. They occur in testes/ovaries (gonadal) and other body sites (extragonadal). There are different types, depending on tumour cell appearance under the microscope, although all are treated identically. Compared with other paediatric tumours, very little is known regarding factors that affect outcome in MGCTs. Studies of genetic changes in tumours may give an indication of the likely response to treatment. We have analysed expression of tiny fragments of genetic code called microRNA in MGCT samples. MicroRNAs act as cancer-forming genes by regulating other genetic code (messenger RNA) which produce proteins that control cell growth/vital functions.
Prospective outcomes: Include identification of common/distinct patterns of microRNA expression between different MGCTs; correlation of these profiles with other MGCT genetic changes and clinical features; comparison with microRNA profiles in adult MGCTs; studying effects of altering microRNA levels in model systems.
Expected benefits: Our findings may allow identification of 'biomarkers' used to help diagnosis and predict outcome in MGCTs as well as suggesting potential targets for new drug treatments.
Research Proposal: MGCTs come from egg- or sperm-producing cells, representing 4% of childhood cancers. They occur in testes/ovaries (gonadal) and other body sites (extragonadal). There are different types, depending on tumour cell appearance under the microscope, although all are treated identically. Compared with other paediatric tumours, very little is known regarding factors that affect outcome in MGCTs. Studies of genetic changes in tumours may give an indication of the likely response to treatment. We have analysed expression of tiny fragments of genetic code called microRNA in MGCT samples. MicroRNAs act as cancer-forming genes by regulating other genetic code (messenger RNA) which produce proteins that control cell growth/vital functions.
Prospective outcomes: Include identification of common/distinct patterns of microRNA expression between different MGCTs; correlation of these profiles with other MGCT genetic changes and clinical features; comparison with microRNA profiles in adult MGCTs; studying effects of altering microRNA levels in model systems.
Expected benefits: Our findings may allow identification of 'biomarkers' used to help diagnosis and predict outcome in MGCTs as well as suggesting potential targets for new drug treatments.
Technical Summary
Aims/objectives: By studying the role of microRNAs in paediatric malignant germ cell tumours (MGCTs) and germ cell development we aim to 1) identify diagnostic tools/prognostic markers 2) suggest targets for the generation of novel therapeutic agents and 3) identify microRNAs that play an important role in vertebrate development.
Background: Paediatric MGCTs occur at several different anatomical sites and are heterogeneous in nature. There are a number of different histological subtypes, but despite this, treatment is almost identical for all cases. They represent 4% of cancers in childhood, but their incidence is increasing and, furthermore, testicular MGCTs are a leading cause of death in young men. Our understanding of MGCT biology lags significantly behind other paediatric solid tumours and leukaemias. MicroRNAs (miRNAs) are short, non protein-coding RNAs that act post-transcriptionally as regulators of gene expression. miRNAs act as oncogenes and tumour suppressor genes in their own right and are reported to be diagnostic and prognostic markers in certain cancers. We propose to study these novel gene regulators in this under-investigated paediatric cancer group.
Design/methodology: Using Exiqon microarray technology we have performed global expression profiling for 585 known human miRNAs, using the UK national collection of paediatric MGCTs as well as control tissues and cell lines. Unsupervised analysis, determined by the expression of 275 miRNAs, demonstrated clustering largely by histological subtype. Supervised clustering analysis identified 127 known miRNAs most differentially expressed between histological subtypes, controls and cell lines (p<0.0001). Current validation of this data using Taqman quantitative real time PCR (qRT-PCR) and in-situ hybridisation (ISH) is in progress. We will further investigate common/distinct patterns of miRNA expression between different subtypes of MGCT and between adult and childhood disease, exploiting the extensive clinical and follow-up data that is available. We will relate these profiles to genomic changes and protein-coding gene expression profiles in the same samples. We will independently validate our findings on separate MGCT samples/tissue microarray. Using bioinformatics (e.g. miRBase) we will identify candidate miRNAs that act as oncogenes or tumour suppressor genes via direct negative regulation of protein-coding genes involved in the pathogenesis of MGCTs.
Scientific opportunities will arise from functional studies looking at the biological effects of over-expression/depletion of these miRNAs in MGCT cell lines. Collaboration on a Xenopus model will allow study of microRNA expression during vertebrate development.
Medical opportunities will arise from identification of key miRNAs that can be used diagnostically and prognostically as clinical tools.
Background: Paediatric MGCTs occur at several different anatomical sites and are heterogeneous in nature. There are a number of different histological subtypes, but despite this, treatment is almost identical for all cases. They represent 4% of cancers in childhood, but their incidence is increasing and, furthermore, testicular MGCTs are a leading cause of death in young men. Our understanding of MGCT biology lags significantly behind other paediatric solid tumours and leukaemias. MicroRNAs (miRNAs) are short, non protein-coding RNAs that act post-transcriptionally as regulators of gene expression. miRNAs act as oncogenes and tumour suppressor genes in their own right and are reported to be diagnostic and prognostic markers in certain cancers. We propose to study these novel gene regulators in this under-investigated paediatric cancer group.
Design/methodology: Using Exiqon microarray technology we have performed global expression profiling for 585 known human miRNAs, using the UK national collection of paediatric MGCTs as well as control tissues and cell lines. Unsupervised analysis, determined by the expression of 275 miRNAs, demonstrated clustering largely by histological subtype. Supervised clustering analysis identified 127 known miRNAs most differentially expressed between histological subtypes, controls and cell lines (p<0.0001). Current validation of this data using Taqman quantitative real time PCR (qRT-PCR) and in-situ hybridisation (ISH) is in progress. We will further investigate common/distinct patterns of miRNA expression between different subtypes of MGCT and between adult and childhood disease, exploiting the extensive clinical and follow-up data that is available. We will relate these profiles to genomic changes and protein-coding gene expression profiles in the same samples. We will independently validate our findings on separate MGCT samples/tissue microarray. Using bioinformatics (e.g. miRBase) we will identify candidate miRNAs that act as oncogenes or tumour suppressor genes via direct negative regulation of protein-coding genes involved in the pathogenesis of MGCTs.
Scientific opportunities will arise from functional studies looking at the biological effects of over-expression/depletion of these miRNAs in MGCT cell lines. Collaboration on a Xenopus model will allow study of microRNA expression during vertebrate development.
Medical opportunities will arise from identification of key miRNAs that can be used diagnostically and prognostically as clinical tools.
Organisations
- MRC Cancer Unit (Lead Research Organisation)
- AstraZeneca (Collaboration)
- EMBL European Bioinformatics Institute (EMBL - EBI) (Collaboration)
- UNIVERSITY OF NOTTINGHAM (Collaboration)
- Southwestern Medical Center (Collaboration)
- McGill University Health Centre (Collaboration)
- The Wellcome Trust Sanger Institute (Collaboration)
People |
ORCID iD |
Matthew Jonathan Murray (Principal Investigator) |
Publications
Bailey S
(2015)
Biallelic somatic SMARCA4 mutations in small cell carcinoma of the ovary, hypercalcemic type (SCCOHT).
in Pediatric blood & cancer
Bailey S
(2014)
Serum microRNA screening for DICER1-associated pleuropulmonary blastoma.
in Pediatric blood & cancer
Collinson K
(2014)
Age-related biological features of germ cell tumors.
in Genes, chromosomes & cancer
Gillis AJ
(2013)
Targeted serum miRNA (TSmiR) test for diagnosis and follow-up of (testicular) germ cell cancer patients: a proof of principle.
in Molecular oncology
Hanning JE
(2013)
Lack of correlation between predicted and actual off-target effects of short-interfering RNAs targeting the human papillomavirus type 16 E7 oncogene.
in British journal of cancer
Hanning JE
(2013)
Depletion of HPV16 early genes induces autophagy and senescence in a cervical carcinogenesis model, regardless of viral physical state.
in The Journal of pathology
Jeyapalan JN
(2011)
Methylator phenotype of malignant germ cell tumours in children identifies strong candidates for chemotherapy resistance.
in British journal of cancer
Muralidhar B
(2011)
Functional evidence that Drosha overexpression in cervical squamous cell carcinoma affects cell phenotype and microRNA profiles.
in The Journal of pathology
Murray M
(2010)
Germ cell tumours in children and adolescents
in Paediatrics and Child Health
Murray M
(2018)
"Future-Proofing" Blood Processing for Measurement of Circulating miRNAs in Samples from Biobanks and Prospective Clinical Trials
in Cancer Epidemiology, Biomarkers & Prevention
Description | Demonstration of feasibility of detecting differential expression of microRNAs in serum from patients at diagnosis of malignant germ cell tumours |
Geographic Reach | Multiple continents/international |
Policy Influence Type | Citation in clinical reviews |
Description | Experimental Cancer Medicine Centres (ECMC) Steering Group: Standardisation of analysis and reporting of circulating miRNAs by RTqPCR |
Geographic Reach | Multiple continents/international |
Policy Influence Type | Membership of a guideline committee |
Description | MaGIC US UK GCT collaboration |
Geographic Reach | Multiple continents/international |
Policy Influence Type | Contribution to a national consultation/review |
Description | Member of NCRI CCL GCT subgroup |
Geographic Reach | National |
Policy Influence Type | Participation in a guidance/advisory committee |
Description | Organised 1st International Extracranial Germ Cell Tumour Conference, Cambridge, September 2019 - this work targeting microRNAs was presented and published in European Urology Supplements |
Geographic Reach | Multiple continents/international |
Policy Influence Type | Influenced training of practitioners or researchers |
Description | 'Novel genetic markers for blood-based diagnosis of common childhood cancers' |
Amount | £44,087 (GBP) |
Funding ID | 11CAM01 |
Organisation | Sparks Charity |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 05/2012 |
End | 05/2013 |
Description | 'Novel genetic markers for blood-based monitoring of treatment response in common childhood cancers' |
Amount | £99,840 (GBP) |
Organisation | Children with Cancer UK |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 09/2012 |
End | 12/2014 |
Description | : 'A clinical dataset and tumour bank for malignant germ cell tumours in young people in the UK' |
Amount | £26,833 (GBP) |
Organisation | Laura Crane Youth Cancer Trust |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 04/2012 |
End | 11/2014 |
Description | ACT-NIHR Cambridge BRC Cancer Research Training Fellowship |
Amount | £49,000 (GBP) |
Organisation | Addenbrooke's Charitable Trust (ACT) |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 02/2014 |
End | 01/2015 |
Description | AGCT1531: A Phase 3 Randomized Trial of Carboplatin vs Cisplatin for Paediatric and Adult Patients with Extracranial Malignant Germ Cell Tumours |
Amount | £1,060,000 (GBP) |
Funding ID | 25632 |
Organisation | Cancer Research UK |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 01/2018 |
End | 01/2026 |
Description | Addenbrooke's Charitable Trust, Cambridge, UK |
Amount | £55,781 (GBP) |
Organisation | Addenbrooke's Charitable Trust (ACT) |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 01/2012 |
End | 12/2012 |
Description | Assessing the clinical utility of blood-based microRNAs for the management of malignant germ cell tumours |
Amount | £32,611 (GBP) |
Organisation | University of Cambridge |
Department | Isaac Newton Trust |
Sector | Academic/University |
Country | United Kingdom |
Start | 02/2016 |
End | 01/2018 |
Description | Astra Zeneca blood-based microRNA project |
Amount | £150,000 (GBP) |
Organisation | AstraZeneca |
Sector | Private |
Country | United Kingdom |
Start | 08/2014 |
End | 08/2016 |
Description | Improving Outcomes for Children and Young Adults with Extracranial Germ Cell Tumours |
Amount | $2,300,000 (USD) |
Funding ID | RG79925 |
Organisation | St. Baldrick's Foundation |
Sector | Charity/Non Profit |
Country | United States |
Start | 02/2016 |
End | 01/2021 |
Description | Targeting oncogenic microRNA clusters in malignant germ cell tumours using tiny locked nucleic acids |
Amount | £197,895 (GBP) |
Organisation | Action Medical Research |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 02/2015 |
End | 01/2018 |
Description | Targeting oncogenic microRNA clusters in malignant germ cell tumours using tiny locked nucleic acids |
Amount | £149,000 (GBP) |
Organisation | Children with Cancer UK |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 01/2015 |
End | 12/2017 |
Title | Highly sensitive qRT-PCR detection method for serum microRNAs in malignant GCTs |
Description | Research method for detecting differentially expressed microRNAs in the serum in human patients with malignant germ cell tumours |
Type Of Material | Technology assay or reagent |
Year Produced | 2011 |
Provided To Others? | Yes |
Impact | From first description in 2011, four further publications have been published in this field (we are authors on three of the five papers) describing utility of this approach in robustly diagnosing these tumours |
Title | Pipeline for analysing circualting microRNAs in malignant germ cell tumours |
Description | A pipeline to quantify serum and cerebrospinal fluid microRNAs for diagnosis and detection of relapse in paediatric malignant germ-cell tumours. Murray MJ, Bell E, Raby KL, Rijlaarsdam MA, Gillis AJ, Looijenga LH, Brown H, Destenaves B, Nicholson JC, Coleman N. Br J Cancer. 2016 Jan 19;114(2):151-62. doi: 10.1038/bjc.2015.429. Epub 2015 Dec 15. PMID: 26671749 |
Type Of Material | Data analysis technique |
Year Produced | 2016 |
Provided To Others? | Yes |
Impact | New clinical trial established in 2016 (AGCT1531) testing this out |
Description | Astra Zeneca blood-based microRNA project |
Organisation | AstraZeneca |
Country | United Kingdom |
Sector | Private |
PI Contribution | Working together to establish blood-based microRNA testing for patients with malignant germ cell tumours - infrastructure, lab skills and knowledge |
Collaborator Contribution | Post-doctoral scientist and consumables for minimum two year period, 2014-2016. |
Impact | Just commenced in September 2014 |
Start Year | 2014 |
Description | Collaboration with Sanger Institute |
Organisation | The Wellcome Trust Sanger Institute |
Country | United Kingdom |
Sector | Charity/Non Profit |
PI Contribution | Shared working in understanding the molecular biology of germ cell tumours |
Collaborator Contribution | RNA sequencing expertise and bioinformatic analyses |
Impact | None as yet |
Start Year | 2019 |
Description | Collaboration with UTSW team, Dallas, US |
Organisation | Southwestern Medical Center |
Country | United States |
Sector | Hospitals |
PI Contribution | Furthering our world-leading science on developing circulating microRNA testing for the clinic |
Collaborator Contribution | Sample, reagent and intellectual contributions to this partnership with shared research outputs |
Impact | Shared publications |
Start Year | 2019 |
Description | Collaboration, EMBL-EBI, Hinxton, Cambridge, UK |
Organisation | EMBL European Bioinformatics Institute (EMBL - EBI) |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Provide raw microarray and next generation sequencing data for analysis, validate results using qRT-PCR etc, jointly analyse results. |
Collaborator Contribution | Bioinformatic support for processing mRNA and miRNA microarray data, next generation sequencing data etc Use of novel bioinformatic tools to integrate mRNA and miRNA datasets |
Impact | Outputs include accepted abstracts for poster and oral presentation at national and international meetings, as well as publications in peer reviewed journals (Cancer Research, Molecular Cancer). |
Start Year | 2008 |
Description | DICER1 mutations in germ cell tumours |
Organisation | McGill University Health Centre |
Department | Department of Medical Genetics |
Country | Canada |
Sector | Academic/University |
PI Contribution | Provision of samples, expertise and manuscript preparation/revision |
Collaborator Contribution | Processing of samples |
Impact | Two publications thus far - Witkowski J Pathol 2013 and Witkowski BJC 2013 |
Start Year | 2012 |
Description | Nottingham collaboration |
Organisation | University of Nottingham |
Department | School of Biomedical Sciences Nottingham |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | Contributing samples, clinical and laboratory expertise, manuscript writing |
Collaborator Contribution | Analysing samples and data analysis |
Impact | Manuscript published in British Journal of Cancer 2011 |
Start Year | 2009 |
Title | Circulating microRNA quantification for malignant germ cell tumour diagnosis and monitoring |
Description | Circulating microRNA quantification for diagnosis and disease-monitoring of patients with extracranial germ cell tumours. The test is being used to correlate with current serum biomarkers and imaging. If successful, it is anticipated that circulating microRNAs will be used to alter clinical management in subsequent trials. |
Type | Diagnostic Tool - Non-Imaging |
Current Stage Of Development | Early clinical assessment |
Year Development Stage Completed | 2019 |
Development Status | Under active development/distribution |
Clinical Trial? | Yes |
Impact | This technology is now embedded in two international RCTs for germ cell tumours - AGCT1531 (NCT03067181) and P3BEP (NCT02582697) |
Title | EpSSG clinical trials |
Description | EpSSG group is planning new European prospective clinical trials of paediatric soft-tissue sarcoma. They wish to consider including diagnostic and disease-monitoring serum microRNAs in their study, and so have asked me to give a presentation at their meeting in Barcelona in December 2013. |
Type | Diagnostic Tool - Non-Imaging |
Current Stage Of Development | Initial development |
Year Development Stage Completed | 2013 |
Development Status | Under active development/distribution |
Impact | Impact awaited. |
Title | Prospective study of serum microRNAs in malignant GCT patients |
Description | Collaborative prospective study of serum microRNAs in malignant GCT patients |
Type | Diagnostic Tool - Non-Imaging |
Current Stage Of Development | Early clinical assessment |
Year Development Stage Completed | 2013 |
Development Status | Under active development/distribution |
Impact | Currently planning to embed within next phase of international clinical trials for patients with malignant GCTs via MaGIC collaboration. |
Description | Lay research update talk at GOSHCC |
Form Of Engagement Activity | A talk or presentation |
Part Of Official Scheme? | Yes |
Geographic Reach | Regional |
Primary Audience | Supporters |
Results and Impact | Talk resulted in interest from research supporters re potential impact of blood-based diagnosis and disease monitoring in children with cancer Impact awaited |
Year(s) Of Engagement Activity |
Description | NCRI/CRUK press release for serum microRNA work in childhood tumours |
Form Of Engagement Activity | A press release, press conference or response to a media enquiry/interview |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | Much interest from research and clinical colleagues re this methodology |
Year(s) Of Engagement Activity | 2014 |
URL | http://www.cancerresearchuk.org/about-us/cancer-news/press-release/2014-11-01-cancer-cell-fingerprin... |
Description | Science Week, Cambridge, UK |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | Yes |
Geographic Reach | Regional |
Primary Audience | Public/other audiences |
Results and Impact | Explained posters to adults, encouraged children to play games designing anti-cancer drugs etc. High level of interest from public and large numbers of people attended the workshop. |
Year(s) Of Engagement Activity | 2008,2009 |
Description | University of Cambridge Press Release - LIN28/let-7 axis in germ cell tumours |
Form Of Engagement Activity | A press release, press conference or response to a media enquiry/interview |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | Interest from media and reported in national press, August 2013 Increased correspondence from patient groups and other health care professionals re the potential for targetting this pathway therapeutically |
Year(s) Of Engagement Activity | |
URL | http://www.cam.ac.uk/research/news/scientists-discover-a-molecular-switch-in-cancers-of-the-testis-a... |
Description | eCancer TV interview |
Form Of Engagement Activity | A press release, press conference or response to a media enquiry/interview |
Part Of Official Scheme? | No |
Geographic Reach | International |
Primary Audience | Professional Practitioners |
Results and Impact | Impact not yet known - recorded at NCRI conference November 2014 No impacts yet |
Year(s) Of Engagement Activity |