manchester cancer research centre research report 2013/14 researc… · welcome to the 2013-14...
TRANSCRIPT
Manchester Cancer Research Centre Research Report 2013/14
Manchester Cancer Research Centre Research Report 2013-14
Manchester Cancer Research CentreThe University of ManchesterWilmslow RoadManchester M20 4BXTel: +44 (0) 161 446 3156www.manchester.ac.uk/mcrc
Founding Partners:
The University of ManchesterOxford Road Manchester M13 9PL Tel: +44 (0) 161 306 6000www.manchester.ac.uk
The Christie NHS Foundation TrustWilmslow RoadManchesterM20 4BX Tel: 0845 226 3000www.christie.nhs.uk
Cancer Research UK Angel Building407 St John StreetLondonEC1V 4ADTel: +44 (0) 20 7242 0200www.cancerresearchuk.org
Copyright © Manchester Cancer Research Centre
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 4 3
ContentsRe
sear
ch R
epor
t
2013
/14
2003
/14
Chair’s Foreword 4 Introduction 5
Ambitious plans for 8 Experimental Therapeutics Andrew Hughes, Jonathan Tugwood, Caroline Dive, Matt Krebs, Emma Dean
Breast Cancer research at the 14 Manchester Breast Centre Tony Howell, Rob Clarke, Gareth Evans
Circulating tumour cell (CTC) research 18 in early stage, resectable non-small cell lung cancer (NSCLC) Caroline Dive, Fiona Blackhall, Phil Crosbie, Rajesh Shah
Heterogeneity in Melanoma 24 Claudia Wellbrock
Translating Cancer Biology into 26 Novel Therapeutics Allan Jordan, Ian Waddell and Donald Ogilvie
Collaboration pays dividends for 32 Leukaemia research Tim Somervaille
Ovarian cancer and angiogenesis 34 Gordon Jayson
Key role for Radiotherapy in 38 personalised approach to cancer treatment Tim Illidge, Catharine West and Nick Slevin
Imaging Science research at the MCRC 42 JamesO’Connor,AdamMcMahon,GeoffParker,
Kaye Williams and Alan Jackson
Biobank building on success 48 Jane Rogan
Author Biographies 54
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 44
Chair’s ForewordWelcome to the 2013-14 Biannual Research Report of the Manchester Cancer
Research Centre (MCRC).
The research work at the MCRC lies at the heart of cancer care not only in
Manchester but both nationally and internationally. Once again we continue to
see great strides forward in all aspects of our work.
The Research Report is a snapshot of some of the impressive progress that has
been made at the MCRC during 2013 and 2014, in terms of both growth and
research output. Even a quick glance at the Report is enough to see that our
scientistsandclinicianscontinuetobepioneersintheirfield,providinginnovative
approaches to the diagnosis and treatment of cancer as well as increasing our
fundamental understanding of the disease at a cellular level.
The work we undertake in the MCRC makes us not only a national beacon of
excellence in cancer research but highlights the fact that we are an internationally
networked Centre that puts into practice our vision of precision medicine.
The last two years have been very successful for the Centre. The MCRC
received renewed and expanded centre status from Cancer Research UK which
will enable us to implement an ambitious plan for further growth, including the
development of a Centre for Biomarker Sciences. This new centre will provide
state-of-the-art facilities for researchers and clinicians, enabling them to
accelerate biomarker sciences and research and bring about a future where
all cancer treatment is tailored to individual patients. New facilities are being
planned that will complement the iconic new MCRC building that is nearing
completion on our Withington site.
The arrival of formidable world-leading researchers has strengthened expertise
within key research areas and enables us to maximise the opportunities we have
in proton therapy and image-guided radiotherapy research. Collaborations and
networkingarekey tosuccessful scientificendeavourandMCRCresearchers
have played their part by driving the development of Centres of Excellence in
Lung Cancer with UCL in London, Prostate Cancer with Queens University,
Belfast and Cancer Imaging with the University of Cambridge. The creation of
these Centres really does recognise the diverse contributions that Manchester
makes across the full cancer research spectrum.
Looking to the future, we are committed to delivering on our promise. We have
anunrivalledopportunityinManchestertoofferlifesavinghopeforpatientswith
cancer by matching the right treatment to the right patient. We aim to become
not just a national beacon of excellence in cancer research, but an internationally
networked centre, and put into practice our vision of precision medicine.
Michael Oglesby
Chair
Manchester Cancer Research Centre
Michael OglesbyChairManchester Cancer Research Centre
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 4 5
IntroductionThe vision of the MCRC is one of precision medicine, where each cancer patient
receives the right treatment at the right time, tailored to his or her individual tumour.
Our aim therefore is to better understand the cellular processes and mechanisms
that drive cancer initiation and growth, in order to identify potential targets for
therapeutics, to develop such novel agents and to evaluate new treatment
approaches in the clinic. This report highlights some key research areas that
are contributing to the discovery and implementation of more personalised
cancer treatments.
Vital to our vision is the Experimental Cancer Medicine team, based at The
Christie’s Clinical Trials Unit. The article by Andrew Hughes and collaborators from
the Clinical and Experimental Pharmacology group highlights the successes of
the Unit and the strengths of Manchester as a location for Experimental Cancer
Medicine research. By having early phase trials co-located on our Withington
site with a wealth of biomarker science expertise, we really are in a position to
drive forward biomarker-driven trials of novel agents. The ambition of Andrew
and his team is formidable, and recent progress gives indications of an exciting
future for Experimental Cancer Medicine at the MCRC.
The Manchester Breast Centre is an important part of the MCRC and its groups
continue to work on various aspects of breast cancer research. The report
by Tony Howell, Gareth Evans and Rob Clarke summarises how their recent
laboratory discoveries relate to the current concept of normal breast biology
and describes clinical studies looking at the prediction and prevention of breast
cancer risk.
Lung cancer is a priority area for the MCRC, and in 2014 we became a Cancer
Research UK Lung Cancer Centre of Excellence, in conjunction with colleagues
at UCL. Earlier diagnosis is key to improving survival rates in this disease, but
we also need to better understand the process of metastasis. The report from
the Lung Cancer group details progress being made in two studies that hope
to improve our understanding of the molecular mechanisms behind metastasis.
Professor Nic JonesDirectorManchester Cancer Research Centre
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 46
Their work is focusing on the potential of circulating tumour cells (CTCs) to provide information on metastatic
behaviour in early stage non-small cell lung cancer (NSCLC). The impressive development of CTC-derived
explant models using CTCs taken from small cell lung cancer (SCLC) patients has now been replicated in
NSCLC and this pioneering approach can hopefully be used to predict risk of disease recurrence.
As in other cancer types, genetic and phenotypic heterogeneity within melanoma must be considered when
developing and applying therapeutic approaches. The report by Claudia Wellbrock summarises her laboratory’s
work looking at cellular subpopulations that contribute to both treatment response and invasion. The group
found, in separate studies, that the factors MITF and TNFα both play a role in treatment resistance and in a
fascinating experiment involving zebrafish, the researchers showed that heterogeneous melanoma cells
worked cooperatively in order to invade surrounding tissue.
A real strength of the MCRC is the experience and facilities within the Drug Discovery Unit, allowing us to
translatelaboratoryfindingsfromourresearchgroupsintopotentialnewsmall-moleculecancertherapeutics.
The report from Donald Ogilvie, Allan Jordan and Ian Waddell focuses on two projects that have seen great
progress over the last two years – those looking at the development of PARG and RET inhibitors.
Collaboration is key to successful research, and a joint project between our drug development team and
clinician scientist Tim Somervaille has resulted in an exciting development in Haemato-oncology. Tim’s
article highlights work exploring the potential of LSD1 inhibition for the treatment of acute leukaemia – early
laboratory work by the Leukaemia Biology group and the Drug Discovery Unit has led to a collaboration with
Oryzon Genomics, a Spanish biotech company, on a phase I clinical trial.
Manchester Cancer Research Centre Research Report
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 4 7
Identifying the hallmarks of cancer is one route to developing new anti-cancer treatments. In the search for
novel targets in ovarian cancer, angiogenesis is the focus for Gordon Jayson and his group. His report details
recent experiments by his group exploring the role played by heparan sulphate in regulating angiogenesis-
related growth factors.
The role of radiotherapy in more personalised treatment approaches must not be forgotten. Indeed, the article
from the Radiotherapy Related Research group summarises work done by Catharine West to investigate
potential biomarkers to predict radiosensitivity, radiotherapy-related toxicity and benefit from hypoxia
modifying agents. In addition, Tim Illidge reports results from several studies using novel immunomodulatory
agentstoincreasetheeffectofionisingradiation.
Our expertise in cancer imaging was recognised in 2013 with the award of a CRUK-EPSRC Cancer Imaging
Centre, in conjunction with Cambridge. Since then, good progress has been made. The article from some
of the Centre’s leading investigators highlights success across a wide portfolio of projects, including the
development of a ‘roadmap’ for the validation of imaging biomarkers.
Following its creation in 2007, the MCRC Biobank has supported research across the Centre through the
collection and provision of high quality patient samples. The report from Jane Rogan and Noel Clarke details
howtheirflexibleapproachtobiobankinghasfacilitatedavarietyofstudiesandenabledManchestertomakea
significantcontributiontonationalresearchinitiatives.
We are entering a new era of cancer research in Manchester and, thanks to the impressive progress over
the last two years, are well placed to deliver on our vision of a personalised approach to treatment for all
cancer patients.
Professor Nic Jones
Director
Manchester Cancer Research Centre
Introduction
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 48
Manchester Cancer Research Centre Research Report
Andrew Hughes, Jonathan Tugwood, Caroline Dive, Matt Krebs, Emma Dean
Ambitious plans for Experimental Therapeutics
Experimental Medicine was defined as one of three core themes
alongside Radiation-Related Research and Lung Cancer in the successful
2008 Manchester Cancer Research Centre (MCRC) Cancer Research UK
Centre application.
This facilitated the MCRC to develop a 2000m2 state-of-the-art Clinical Trials
Unit comprising 31 beds/treatment chairs, 6 inpatient beds, 5 outpatient suites
and a large sample collection laboratory. The Unit has supported 66 Phase I
and 183 Phase II clinical trials, involving 3,202 patients in experimental studies
since its opening in 2010. It was ranked as a ‘forefront’ UK Experimental Cancer
Medicine Centre (ECMC), recruiting 25% of all UK patients into ECMC clinical
trials from the largest catchment population (3.2M people) for a cancer centre
in England and Wales with over 14,000 new patients/annum. The Clinical Trials
Unit was built with the aim that it would become one of the largest centres
for experimental cancer medicine trials worldwide. We remain committed to
this target, which will provide clear evidence that The Christie and the broader
MCRC is a leading cancer centre.
The Experimental Cancer Medicine team, located within the Clinical Trials
Unit, conducts early phase clinical trials comprising first-in-human and first-
in-combination studies with the objective of providing a recommended dose
and schedule for further (Phase II) testing. The Experimental Cancer Medicine
team also conduct ‘Regulatory Clinical Pharmacology’ trials. These trials often
comprise the majority of text in the drug prescribing information and the
majority of clinical study reports in a submission dossier. These studies seek to
characterise the impact of food (food interaction studies), organ impairment
(renal and hepatic studies), concomitant medication (drug-drug interaction
studies), formulation changes (bioequivalence and bioavailability studies)
upon the exposure (pharmacokinetics) of the drug; and characterisation of
absorption, distribution, metabolism, elimination (ADME) of the drug.
The Experimental Cancer Medicine team within the Clinical Trials Unit is
ideally located next to the cancer discovery and translational laboratories
within the Cancer Research UK Manchester Institute (CRUK MI). Other key
translational platforms have also been developed within the MCRC including (i)
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 4 9
Ambitious plans for Experimental Therapeutics
The Wolfson Molecular Imaging Centre, which together with colleagues from
Cambridge recently attained CRUK-EPSRC Cancer Imaging Centre status and
is well equipped to deliver imaging biomarkers for application in experimental
therapeutics cancer clinical studies (PET, MRI, CT, MRS); (ii) Clinical and
Experimental Pharmacology (CEP) laboratories - one of the best laboratories
world-wide in cancer biomarker discovery, validation and development within
the CRUK MI (iii) an extensive and successful biobanking infrastructure that
can capitalise on the very high patient population. Planned delivery of an
Integrated Procedures Unit from 2016 will further augment the ability to deliver
translational clinical studies, with the provision of radiology guided biopsies
and procedures for bone marrow aspiration and surgical biopsies (breast, skin,
melanoma, sarcoma, renal, prostate, bladder, cervix, endometrium, ovarian,
vulval, vaginal, colorectal, anal, and peritoneal).
We now have an enormous opportunity to be a major international centre
for experimental cancer medicine providing access to novel therapies for
a significant proportion of patients. We aim to become a ‘go-to’ centre for
scientificallydrivenandbiomarkerinformedtrials,tobeamagnetforworld-class
staff,tobeatrainingbeaconinexperimentaltherapeutics,andbyimplementing
the biomarker expertise from CEP in the clinic, to become a leader in ‘liquid
biopsy’ precision medicine with real-time clinical trial data acquisition that
enables adaptive decision making in Phase I trials and to work in partnership
with other centres to conduct basket/bucket trials.
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 410
Ourgoaloverthenext fewyears istofully realisethisopportunity: tobefirmlyestablished inthetopthree
UnitsinEuropewithinthenextfiveyearsandinthetopfivegloballywithinthenexttenyears.Thiswillrequire
a5-10foldincreaseinthenumberofearlyphasetrialsoverthenextfiveyearsanda10-20foldgrowthover
the next ten years in the number of patients enrolled into such trials; and in the overall scale of operations,
without compromising the high quality of patient care currently delivered. Our long-term global aspiration will
requireevenfurtherinvestmentandtheestablishmentofeffectiveandintegratednetworkswithothermajor
therapeutic units especially in the UK.
Wehavetakenourfirsttangiblegrowthsteps.InJuly,twoseniorlecturers-DrMattKrebsandDrSaeedRafii-
were appointed alongside Dr Emma Dean, who was appointed as Senior Lecturer in the experimental cancer
medicines team in 2012. Emma has pursued postgraduate study in human pharmacology and together with
ProfessorMalcolm Ranson, who retired earlier in 2014, has administered over 170 different experimental
cancer medicines since 2007. Dr Matt Krebs undertook his PhD in circulating biomarkers in CEP and his research
interestisinnon-smallcell lungcancer.DrSaeedRafiijoinedusfromtheRoyalMarsdenHospital,whichhas
a thriving experimental cancer medicine team within the Drug Development Unit that enrols 300-400 new
patients into experimental cancer medicine studies each year. Saeed’s insights into running a department
of this scale will be invaluable as we scale up. From April 2015, Professor Andrew Hughes will be joining the
experimental cancer medicine team as Strategic Director to drive investment and growth. Andrew has held a
joint appointment with The University of Manchester as Chair in Experimental Therapeutics since 2008 and
has been vice-president for early oncology clinical development within AstraZeneca. Andrew has commenced
workwiththeExperimentalCancerMedicineteamtodefinethetrajectoryforgrowthandinvestment.
Manchester Cancer Research Centre Research Report
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 4 11
Sincetheirappointments,SaeedRafii,MattKrebsandEmmaDeanhavebroughtsevennewtrialstopatients
includinganovelATR inhibitor,withTheChristieenrollingthefirstUKpatienttothefirst-in-humanstudy in
November 2014. Other agents include a TOR1/2 inhibitor and a tyrosine kinase inhibitor which not only
inhibits theEGFRmutated receptor (as thefirstgenerationEGFR-TKIssuchas IressaandTarceva)but the
resistance mutation (T790) which develops in some 50% of patients with non-small cell lung cancer. The orally
administered drug is set to submit its registration dossier in 2015 with the experimental cancer medicines
unit conducting three regulatory clinical pharmacology studies to support this submission. Discussions are
ongoing with sponsors for a further seven trials.
In addition to increasing the number of new experimental cancer medicines reaching patients, the
experimental cancer medicine team, working closely with CEP, has also commenced work to build a ‘liquid
biopsy precision medicine’ capability. This maps to the research themes of the CRUK MI and the University’s
Institute of Cancer Sciences. Several major cancer centres across the world are routinely performing DNA
sequencingontheirpatients’tumourtissuetoinformtreatmentstratification-tryingtomatchtheobserved
molecular aberration to a targeted therapy. Recognising that we are behind other cancer centres, we have
electedto focusuponestablishingtheclinical researchcapability formultiplexmolecularprofilingonblood borne tumour derived material (circulating tumour cells (CTCs) and circulating tumour DNA (ctDNA)). Patients
will therefore simply need to provide a blood sample (a ‘liquid’ or virtual biopsy) rather than having to undergo
a tumour tissue biopsy with associated morbidity and mortality. Already treatment decisions are being made
uponthemolecularaberrationsfound inctDNA. InSeptember2014, Iressa-firstdosedtopatients inThe
Christie by Professor Ranson - received approval from the EU licensing authorities, meaning that patients with
non-smallcelllungcancercanbetreatedwithIressauponfindingaEGFRmutationinctDNAwithouthaving
to proceed to a tumour biopsy. The ability to characterise the molecular phenotype of a cancer based upon a
‘liquid biopsy’ will be a critical element in repositioning experimental cancer medicine from being considered
only after all standard of care options have been exhausted (and thus typically an end-of-life experience for
patients) to being considered earlier in the treatment pathway when drugs available under an expanded clinical
trialsportfoliocanbematchedtoputativetumourmoleculardrivers identifiedthroughmultiplexmolecular
profilingfromabloodsample.
The Manchester Cancer Research Centre Conference: Harnessing Apoptosis
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 412
Manchester Cancer Research Centre Research Report
The experimental cancer medicine team has now developed a clinical protocol to allow DNA sequencing for
all patients being considered for an early phase clinical trial, aiming to match relevant experimental agents
with individual genetic aberrations. The clinical trial, known as TARGET (Tumour chARacterisation to Guide
Experimental Targetedtherapy), isduetoenrol itsfirstpatient inearly2015.Thepatientconsentformand
patient information sheet were recently recognised by Independent Cancer Patients’ Voice as exemplars in
theirfield.WehavealsosetupasteeringboardforthisprojectcomposedofMCRCmultidisciplinaryexpertise,
inclusive of The Christie NHS Foundation Trust Research and Development team, the University’s Institute
of Cancer Sciences and CRUK MI clinical and translational scientists, the regional genetics laboratories at
St Mary’s within Central Manchester University Hospitals NHS Foundation Trust, and linked to the ongoing
Integrated Procedures Unit project development.
Samples from this TARGET precision medicine clinical trial will be sent to the CEP laboratories for analysis. The
CEP group, headed by Professor Caroline Dive, comprises over 50 scientists and includes teams dedicated to
high quality non-clinical mechanistic cancer research, the development and implementation of nucleic acid
biomarkers, and a team dedicated to the characterisation of circulating biomarkers in ‘liquid biopsy’ samples
from clinical trials and experimental medicine projects, under Good Clinical Practice (GCP) standards and
regulations. Analysis of clinical trials samples to GCP involves teams of project managers, laboratory-based
analysts and a dedicated Quality Assurance group to oversee GCP compliance. CEP currently has 60 clinical trial
and experimental medicine projects in its portfolio, with another 13 currently in the planning stage. The portfolio
focuses on major cancer disease areas with emphasis on lung, melanoma, colorectal, pancreatic and prostate;
sponsorship and funding includes collaborative relationships with seven major international pharmaceutical
companies, UK NHS Hospital Foundation Trusts, UK universities and UK and European research councils.
ThecornerstoneoftheCEPgroup’seffort istheexperimentalcapabilitythatunderpinstheclinicaltrialand
experimental medicine project activity. Importantly, we have a portfolio of 96 biomarker assays validated to
GCP standards, aligned to CTC enumeration and characterisation, protein biomarker analysis by single-plex
andmulti-plexELISA,andgenomicanalysisoftumourbiopsiesandofctDNA.CTCresearch istheflagship
activityofCEP.Wehaveestablishedcapability forCTCenumerationandstainingforspecificmarkersusing
the FDA-approved Veridex CellSearch platform, and also using the Isolation by Size of Epithelial Tumour
Cells (ISET) approach. In addition, to complement existing platforms we are developing several alternative
approaches in collaboration with service provider organisations, to develop the capability to isolate CTCs using
parameters such as cell size, conformation, and isolation of live cells. In conjunction with new CTC isolation
platforms, we are developing the capability to isolate and characterise single CTCs. This is being led by deputy
CEP director Dr Ged Brady and his nucleic acid biomarker team. As well as single CTC isolation using the
DEPArray™ instrument, Ged’s team is optimising techniques for whole genome DNA analysis, plus RNA and
miRNA analysis, from single isolated cells. Preclinical pharmacology in CEP, led by Dr Chris Morrow, has been
developing unique mouse models for lung cancer (termed CDX) based on implantation of isolated CTCs from
patients. This has been highly successful, and the development and early characterisation of these unique CDX
models was the subject of a Nature Medicine publication in June 2014. The models, developed from CTCs from
small cell lung cancer patients, recapitulate patient responses to standard chemotherapy. The CDX models are
now the subject of newly-initiated studies with a number of pharmaceutical companies and academic partners
to assess efficacy of targeted therapies alone and in combinationwith standard of care drugs. Promising
therapies will be translated to early clinical trials in SCLC patients within the CRUK Lung Cancer Centre of
Excellence in collaboration with Dr Fiona Blackhall from Manchester and Dr Martin Forster from UCL.
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 4 13
The Manchester Cancer Research Centre Conference: Harnessing Apoptosis
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 414
Manchester Cancer Research Centre Research Report
Tony Howell, Rob Clarke, Gareth Evans
Breast Cancer research at the Manchester Breast Centre
The major aim of the Manchester Breast Centre is to translate discoveries from
the laboratory to the clinic in order to improve breast cancer risk estimation,
treatment and prevention.
The laboratories associated with the MBC are situated in the Paterson building
in Withington and within the Faculty of Life Sciences on the University’s main
campus. Clinical studies and trials occur at The Christie, the Nightingale and
Genesis Prevention Centre at University Hospital of South Manchester and
throughout the Cancer Network.
Breast cancer is thought to arise from a terminal-duct lobular unit (TDLU) in one
of the 10-20 lobes of the breast. Each lobule comprises basal myoepithelial and
luminal cells with their corresponding progenitor/stem cells. Luminal progenitor
cells are thought to be the predominant site of origin of the majority of breast
cancers. Stromal cells includingfibroblasts, fat, immuneand vascular cells lie
outsidetheepithelialbasementmembranebutinfluenceepithelialgrowthand
transformation. Greater understanding of the interaction between cell types
and systemic factors is likely to lead to better treatments and prevention of the
disease. In 2014 the majority of MBC publications related to mammary stroma
and mammographic density, stromal and stem cell signalling and aspects of risk
estimation and prevention of breast cancer.
Mammary stroma and mammographic density
The mammary stroma is the non-fat part of the breast and comprises non
cellular structures; predominantly collagens, proteoglycans and cellular
elements including fibroblasts, immune and vascular cells. During puberty
mammary ducts extend throughout the breast mainly associated with stroma.
The degree of ductal spread and branching is related to density of the breast,
and high mammographic density is a strong risk factor for breast cancer.
Potential control mechanisms for the degree of branching were reported by
researchers from the Wellcome Trust Centre for Cell-matrix Research. The
group demonstrated fibroblast growth factor (FGF) signalling from stromal
cells tobreastepithelialstemcells;firstbranching initiationoccursunderthe
influenceofFGF10andductalelongationthenoccursviathereleaseofFGF2.It
appears that both growth factors signal through FGFR2 receptors on epithelial
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 4 15
cells. The degree of branching is reduced by an epithelial tyrosine kinase called
Sprouty which may be regarded as a tumour suppressor. Thus the FGF-Sprouty
pathway may regulate the degree of branching and this risk of subsequent
breast cancer.
Twin studies indicate that about two thirds of mammographic density is
hereditary. The other third varies in relation to breast cancer risk factors;
late age of first pregnancy andHRT increase densitywhereas it ismarkedly
reduced in some women treated with tamoxifen. Recently Michael Lisanti and
hiscolleaguesdemonstrated,usinggeneexpressionanalysis,thatfibroblasts
derived from high density breasts had some features of pro-inflammatory
cancer associated fibroblasts including the activation of the JNK1 stress
pathwayassociatedwithinflammationandfibrosis.Wehaveshownpreviously
that tamoxifen reduces risk of breast cancer only in the 50% of women who
have a >10% reduction in mammographic density. The discovery of the
importance of the JNK1 pathway suggests inhibitors of this pathway may be
useful for non-responders to tamoxifen. Another indication of altered stromal
interactions was detected in what is thought to be the earliest lesion on the
pathway to breast cancer, columnar cell hyperplasia. In this lesion we showed
upregulation of the miR 132 in the stroma and downregulation of let-7c in the
epithelium. Let-7c is known to reduce ERα and may be one explanation of the
high proportion of ER+ cells in these lesions.
Epithelial stem/progenitor cell signalling
Since tumours are thought to arise from stem/progenitor cells it is important to
further elucidate this mechanism in order to develop new targets for therapy.
Studies by MBC members have given insight into the regulation of normal
epithelial stem/progenitor proliferation, and how this may be disrupted during
the development of ductal carcinoma in-situ and invasive breast cancer. We
Breast Cancer research at the Manchester Breast Centre
Figure: Current concept of the organisation of the normal breast related to MBC discoveries in 2014. Stromal fibroblasts produce factorswhichaffecttheactivityofepithelialcells and may be activated in dense breasts. Basal progenitor cells give rise to myoepithelial (contracting) cells. Luminal progenitors give rise to sensor cells which contain oestrogen (ER) and progesterone (PR) receptors and respond to circulating oestrogen and progesterone respectively. Alveolar progenitors are found in pregnancy and give rise to milk producing and other cells. Recent work indicates thatstromalcellsinfluenceepithelialprogenitors via stromal derived growth factor (SDF-1) interacting with CXCR-4 receptors and stromal fibroblastgrowthfactors(FGF2&10)interacting with FGFR2 receptor on the epithelial progenitors. Progenitors also interact with stromal integrins which activate focal adhesion kinase (FAK) and downstream pathways. It is thought that basal and luminal progenitors arise during development from primordial stem cells.
1. Duct reconstruction to show lobar structure.
2. Breast of 18 year old sectioned.
3. TDLU Terminal Duct-Lobular Unit showing ductules.
4. Electron micrograph of a cross section of a ductule
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 416
demonstrated that the cytokine receptor CXCR-4 is up-regulated on progenitor cells of the breast. Treatment
withitsstromalactivator(StromalDerivedFactor1-SDF1)hadadualeffect:therewasdown-regulationof
normal progenitors but up-regulation of tumour cell progenitors, a process which could be inhibited by the
bicyclam CXCR-4 inhibitor, AMD 31000, now in clinical development.
Progenitor cells express integrins which sense components of the mammary stroma. For example α6β4
integrin is the cell surface receptor for stromal laminin. In collaboration with a Portuguese group we have
demonstrated that P-cadherin, responsible for cell-cell adhesion, is up-regulated in basal-like mammary
tumours and, in turn, causes down-regulation of α6β4 integrin, increased tumour cell motility and activation
of down stream focal adhesion kinase (FAK). The activation of P-cadherin may explain the increased invasive
properties of epithelial stem cells observed in response to oestrogen.
Analysis of the proteins regulated in tumour stem cells indicated up-regulation of mitochondrial proteins.
Mitochondria require L-lactate and ketone bodies as fuel substrates which are taken up into the cell by
monocarboxylate transporters.Wehave recently shown that specific inhibitionof lactate transport inhibits
stem cell proliferation in oestrogen receptor positive and negative cell lines and suggests the importance of
inhibitors of MCT, such as AZD 3965, entering clinical development. More recently a separate form of stem/
progenitor cell for alveolar milk producing cells was described, the proliferation of which is, in part, controlled by
the transcription factor RUNX2 and B3 integrin. The relevance of the alveolar stem cell to the development of
human breast cancer remains to be discovered.
We have demonstrated that focal adhesion kinase (FAK) is also up-regulated in the progenitor cells of
aggressive forms of human ductal carcinoma in situ (DCIS) and is predictive for relapse and invasion of this
Manchester Cancer Research Centre Research Report
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 4 17
Breast Cancer research at the Manchester Breast Centre
DCIS subtype. FAK inhibition reduces progenitor cell growth and it is planned to test this inhibitor in the clinic.
Lapatinib also reduces DCIS progenitor cell growth and thus may also be used to prevent relapse in patients
with high risk DCIS. In a collaborative study, the mTOR inhibitor everolimus was shown to be highly active when
combined with endocrine therapy for advanced disease. New inhibitors, hopefully with reduced toxicity, are
being developed which could be used for adjuvant therapy and for prevention of DCIS relapse and possibly for
breast cancer prevention.
Recently, there has been an increasing interest in the development and characterisation of patient-derived
tumour xenograft (PDX) models for cancer research. PDX models mostly retain the principal histologic and
genetic characteristics of their donor tumour and remain stable across passages. These models have been
shown to be predictive of clinical outcomes and are being used for preclinical drug evaluation, biomarker
identification,biologicstudies,andpersonalisedmedicinestrategies.
Risk prediction and prevention of breast cancer
Members of the MBC published an analysis of the gaps in our knowledge concerning breast cancer risk
prediction and prevention. The clinical gaps included how to improve risk estimation, new chemopreventative
agents and optimal measures for lifestyle intervention. The laboratory gaps included the mechanism of the
effectsofpregnancy,energyrestrictionanddensityonthebreastandwhy,insomewomen,thereisalackof
involution of the breast after the menopause.
The discovery of the BRCA1 and BRCA2 genes revolutionised the management of women at high risk of breast
cancer. We reported that the uptake of gene testing was doubled after the publicity surrounding Angelina
Jolie’s BRCA positive gene test and subsequent bilateral risk reducing mastectomy. We, and others, have
demonstratedtheeffectivenessofriskreducingsurgeryforpreventingbreastcancerinwomenatveryhigh
riskandalsothatscreeningwithMRIisalsohighlyeffectiveinthisgroupofwomen.Preciseindicatorsofriskof
breastcancerinBRCA1/2carriersareimportantformanagement.Wereportedupdatedfiguresonpenetrance
thatindicatethatitisincreasingovertheyears.Penetrancemaybemodifiedbytheeffectsofsinglenucleotide
polymorphisms found in whole genome breast cancer association studies. However, attempts at selection of
candidatemodifiergenesandtelomerelengthdonotmodifyBRCA1/2andBRCA2penetrance.
The risks of women referred to the Family History Clinic but who are non-gene carriers can be accurately
predicted by using degree of family history and modifying this prediction by adding hormonal risk factors such as
ageoffirstpregnancy.However,mostwomeninthegeneralpopulationarenotawareoftheirrisks.Wetherefore
evaluated whether risk could be assessed in women at screening by mammography (PROCAS: PRediction Of
Cancer At Screening). Over 54,000 women entered the study which demonstrated that approximately 10%
of women are at moderate or high risk of breast cancer. We have preliminary evidence that more precise risk
estimation is possible if the mammographic density and SNPs are added to a standard model. Further work
is required to determine whether such models can be used in the National Breast Screening Programme to
determine the optimal interval for screening and the use of preventative measures.
Studies reported this year indicate progress in the endocrine prevention of breast cancer. Aromatase inhibitors
(AIs) lower oestrogen concentrations and have been shown to be superior to tamoxifen for the prevention of
systemic relapse after surgery for breast cancer. Now, in a study reported by our group earlier this year (the
IBIS-II),theAIanastrozole,whengivenforfiveyears,preventsbreastcancertoagreaterextentthantamoxifen
witharelativelylowsideeffectprofile.HoweveranimportantsideeffectofAIsisreductionofbonedensitybut
now we have shown that this can be abrogated by the bone sparing bisphosphonate once-weekly. Tamoxifen,
alsogivenforfiveyears,isnowacceptedbyNICEforbreastcancerprevention.Withlongerfollowupwehave
shownthatthepreventativeeffectcontinuesforupto20years(theIBIS-Itrial)buttheuptakeoftamoxifenin
women at risk of breast cancer remains relatively low.
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 418
Manchester Cancer Research Centre Research Report
Caroline Dive, Fiona Blackhall, Phil Crosbie, Rajesh Shah
Circulating tumour cell (CTC) research in early stage, resectable non-small cell lung cancer (NSCLC)
A collaboration of the early stage lung cancer and biomarker theme researchers
of the MCRC Lung Group aims to improve knowledge of metastasis.
Lung cancer is the most common cause of cancer-related mortality worldwide
(~1.4 million deaths/year). Five-year survival in the UK is less than 10%. The
most common pathological subtype, non-small cell lung cancer (NSCLC),
accounts for 85% of cases. Early diagnosis and surgical resection of NSCLC can
produce long-term survival (5-year survival: Stage I 58-73%, Stage II 36-46%,
Stage IIIA 24%). However, tumour recurrence occurs in 50% of cases indicating
micro-metastatic disease present at surgical resection undetected by current
stagingstrategies.Recurrenceriskpeaksduringthefirsttwoyearsaftersurgery
most commonly occurring at distant sites when it is almost universally fatal.
Platinum-based adjuvant chemotherapy improves survival by approximately 5%
inpatientswithstageII-IIIA,butbenefitinstageI,where5-yearmortalityis27-
42%, is unproven. To improve outcomes from lung cancer we need to increase
our understanding of the molecular mechanisms of metastasis in early stage
disease to exploit for therapeutic control: this may reveal new drug targets to
pursue in trialsof novel adjuvant therapies.Themore accurate identification
of patients most at risk of recurrence would also enable current management
protocols to be optimised.
In 2014, Dr Phil Crosbie, Senior Clinical Lecturer at The University of Manchester
and Honorary Consultant Respiratory Physician at University Hospital of South
Manchester NHS Foundation Trust (UHSM), in partnership with Professor
Caroline Dive and Dr Fiona Blackhall, secured funding from The Roy Castle
Lung Cancer Foundation (RCLCF) (~£26k) and the Moulton Charitable
Foundation (~£226K) for the conduct of two innovative clinical trials focused
on the detection and study of the molecular biology of circulating tumour cells
(CTCs) in early stage NSCLC. This work closely maps key research objectives of
CancerResearchUK’sfirstLungCancerCentreofExcellenceand isbased in
Professor Caroline Dive’s Clinical and Experimental Pharmacology group (CEP)
at the CRUK Manchester Institute (MI). Here we describe the rationale behind
both studies and how increasing our knowledge of CTCs will further shape our
understanding of this deadly disease.
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 4 19
An investigation of a novel CTC detection device in NSCLC.
CTCs are rare cells that can be detected in blood which are thought to form a
critical step in the development of metastatic disease. Evidence for the tumour
initiating potential of CTCs has been recently shown in xenograft models of
breast and, in Manchester, in small cell lung cancer. The potential for CTCs to be
used as prognostic and predictive biomarkers in advanced lung cancer has been
demonstrated in a number of studies but their application in early stage disease
has been limited by low sensitivity of detection. Addressing the challenge of low
detection is a critical area of CTC research and fundamental to the study of
CTCbiology.Theprimaryaimofthisfirststudyistodeterminewhethermore
CTCs can be detected in the peripheral blood of patients with lung cancer
using a novel CTC detection platform (CellCollector™, GILUPI) compared to
the current ‘gold standard’ (CellSearch®; Janssen Diagnostics). CellSearch®
automatically enriches CTCs by immunomagnetic selection of epithelial cell
adhesion molecule (EpCAM) expressing cells in blood (7.5ml), followed by
semi-automated morphological and immunofluorescent categorisation and
enumeration. CTCs detected in this way have been associated with a worse
prognosis in several solid tumour types including lung cancer. We previously
demonstratedthatinadvancedNSCLC,patientswith≥5CTCs/7.5mlbloodhad
a significantly poorer progression free andoverall survival, however ≥2CTCs
were seen in only 32% stage IV and rarely in stage III disease. Dr Crosbie led a
pilotinearlystageNSCLCthatdemonstrated≥2CTCs/7.5mlperipheralblood
Circulating tumour cell (CTC) research in early stage, resectable non-small cell lung cancer (NSCLC)
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 420
Manchester Cancer Research Centre Research Report
inonly2/34(6%)patients.Weconcludedthatthismayreflectlackofassaysensitivityduetodown-regulation
of EpCAM and cytokeratins during epithelial to mesenchymal transition (EMT) as well as lower prevalence of
CTCs in early disease.
CellCollector™ is a novel CE approved medical device developed by German company GILUPI GmbH. The
device is a functionalised guidewire labelled with antibodies to EpCAM, which is designed to isolate EpCAM+
CTCs from peripheral blood through specific antigen-antibody binding. However, rather than analysing a
small volume of blood, the wire is placed through a standard intravenous cannula into a peripheral vein for a
periodof30minutes;thisresultsinasignificantlyincreasedsampledbloodvolume,estimatedtobe1.5to2
litres,or≈200timesthebloodvolumesampledbyCellSearch®(7.5ml)potentiallyincreasingCTCdetection
significantly.WewillexaminethisdeviceinpatientswithearlystageNSCLCandwillalsoexplorethefeasibility
of downstream molecular characterisation of CTCs.
Does the tumourigenicity of CTCs in early stage NSCLC predict disease recurrence?
It has been assumed for most tumour types that CTCs are the mediators of metastatic spread, but until
recently this assumption had not been formally tested. Using blood from patients with small cell lung cancer
(SCLC), where CellSearch® detected CTCs are relatively abundant, MCRC researchers from Professor Dive’s
CEP group, the University and The Christie demonstrated that CTCs enriched from SCLC patients routinely
form tumours in immune-compromised mice (Figure1A), termed CTC explant models or CDX. Histological
analysis demonstrated CDX were SCLC (Figure 1B) and genetic comparison of the CDX and single CTCs
isolatedfromthesamepatientconfirmedCDXsarederivedfrompatientCTCs(Figure1C).
Figure 1A. CDX derived from SCLC CTCs enriched from blood samples from two patients (patients 2 and 4).
Figure 1B. Comparison of IHC staining of patient 2 pleural fluidor patient 4 diagnostic biopsy and corresponding CDX.
Figure 1C. Whole genome sequencing of CDX2 revealed a c.440T>G transversion in TP53. Sanger sequencing on 6 single CTCs isolated from patient 2 demonstrated the same transversion, which was absent in the leukocyte sample, confirming acancer cell somatic mutation.
Figure 1A.
Figure 1C.
Figure 1B.
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 4 21
Circulating tumour cell (CTC) research in early stage, resectable non-small cell lung cancer (NSCLC)
More recently in a proof of concept experiment, a CDX was successfully generated from blood taken from
apatientwithadvancedNSCLC.Pathologicaland immunologicalprofilingof theCDXmatchedtheprimary
tumour (Figure 2). Dr Crosbie’s focus will be to determine whether CTCs enriched from patients with early
stage NSCLC form CDX and to correlate CDX generation with clinical outcome.
This study will recruit patients undergoing surgical
resection of early stage NSCLC at UHSM where in excess
of 400 resections a year are performed. Dr Crosbie has, in
collaboration with Mr Rajesh Shah, lead Thoracic Surgeon
at UHSM, explored pulmonary vein sampling as a method
for increasing CTC detection in early stage NSCLC. The
pulmonary veins drain blood directly from the lungs; proximity
to primary tumour and blood draw prior to capillary bed
filtering,reportedtoremove90%ofCTCs,makespulmonary
vein sampling at surgery potentially advantageous. Previous
studies (using a variety of CTC enrichment methods) have
reported pulmonary vein CTCs in 18-96% of patients with
the presence of CTCs variably associated with prognosis.
At UHSM blood is taken prior to tumour manipulation
or vessel ligation to minimise artificial elevation in CTC
number. Pilot data has demonstrated pulmonary vein CTCs
(CellSearch®) inmorepatients(≥2CTCs:pulmonary14/34,
Figure 2. H&E staining of biopsy from the NSCLC patientwhose CTCs gave rise to CDX and of the derived CDX (top images). TTF1 and cytokeratin staining of the NSCLC CDX tumour (bottom images).
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 422
Manchester Cancer Research Centre Research Report
41% vs. peripheral vein 2/32, 6%; p=0.001) and in higher numbers (pulmonary 0-3093 vs. peripheral vein 0-4
CTC/7.5ml; p=0.002) than matched peripheral blood (Figure 3).
Pulmonaryveinbloodwillbetakenatsurgery,depletedofredandwhitebloodcellsandimplantedintotheflank
of an immune-compromised mouse. A matched tumour biopsy will be implanted into a second mouse. The
implantation of tumour biopsies to establish patient derived xenografts (PDX) is a well-described technique
that preserves tumour architecture and maintains tumour-stromal interactions allowing assessment of
tumour biology and treatment responses. In NSCLC, the successful establishment of PDX models has been
reported in 25-46% of cases; although PDX generation was linked to disease recurrence post-resection in one
study, the other studies show no correlation with clinico-pathological factors. Once established, response to
chemotherapy in PDX models may predict response in a clinical setting. However, PDX, established using small
singlesitetumourbiopsiesmaynotreflectthefullextentofintra-tumourheterogeneity,postulatedtobean
important driver of disease progression, treatment resistance and a risk factor for disease recurrence after
surgery. It therefore follows that PDX may not be representative of the cells responsible for distant metastatic
spread and disease recurrence in resected early stage patients. On the other hand, the tumourigenicity
(definedbyCDXgeneration)inmiceofcellsthathavealreadyinvadedthroughtissueandintravasatedasCTCs
in the patient may be a much better predictor of a patient’s recurrence risk, the hypothesis that will be tested
in this study.
Figure 3. Pulmonary vein sampling at the University Hospital of South Manchester and an image of a CellSearch detected CTC and WBC. CellSearch CTC enrichment is performed automatically by immunomagnetic selection of EpCAM (epithelial cell adhesion molecule) expressing cells, followed by semi-automated morphological and immune-fluorescent categorisation. Definition of a CTC: EpCAM and cytokeratin (CK; 8, 18 and 19) co-expression,4',6-diamidino-2-phenylindole (DAPI) positive nuclear staining and negative white blood cell marker (CD45) expression.
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 4 23
Circulating tumour cell (CTC) research in early stage, resectable non-small cell lung cancer (NSCLC)
As a result of this study, we hope to be able to stratify early stage NSCLC patients according to risk of recurrence
whomaybenefitfromadjuvantchemotherapy,administeredatpointofconfirmedtakeoftheirderivedCDX
and/or PDX. These patients would also be subjected to more intensive follow up. Our ultimate goal is to identify
amarkerwithinblood to tailor adjuvant treatment topatientswho shouldderivemostbenefit.To achieve
this, the molecular landscape of CDX/PDX will be investigated using next generation sequencing platforms.
For patients whose CTCs and/or resected tumour fragment generated CDX/PDX respectively, a comparison
at the genomic level will also be made with CTCs isolated at resection, resected primary tumour and, when
available, recurrent tumour biopsy. By determining the response of CDX/PDX to chemotherapy regimens, we
hopetoselectaspecificandpersonalisedtherapyregimenforpatientsatpointofdiseaserecurrence.The
datagenerated fromthisstudywillalsocontributetotheongoingdrugtargetdiscoveryefforts forNSCLC
led by Dr John Brognard and Dr Michela Garofalo and work in drug discovery led by Dr Donald Olgivie at the
CRUK MI.
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 424
Manchester Cancer Research Centre Research Report
Claudia Wellbrock
Heterogeneity in Melanoma
The genetic and phenotypic heterogeneity found within melanomas is an
important feature to consider in the development of melanoma therapies,
because the communication between melanoma cell subpopulations can
impact on the efficacy of cancer drugs. Dr ClaudiaWellbrock’s laboratory is
investigating how intra-tumour signalling amongst different cell populations
controls melanoma growth and therapy response.
Our work has revealed that within a heterogeneous tumour there are distinct
subpopulations of melanoma cells that respond differently to the currently
applied MAP-kinase pathway targeting therapies. These subpopulations can be
distinguishedaccordingtotheirMITFexpressionstate.MITF,alineagespecific
transcription factor, is a crucial phenotype determinant and its expression is
distinctively heterogeneous throughout tumours. We have shown that MITF
confers resistance to MAP-kinase pathway therapy in melanoma. We found that
MITF is up-regulated in patients’ melanomas during treatment with MAP-kinase
pathway inhibitors. We discovered that this up-regulation is based on both cell
autonomous processes and signalling from the immune microenvironment.
Figure A heterogeneous tumour consisting of cells with a differentMITF expression state (high:red; low:green).
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 4 25
We have discovered that immune cells, in particular macrophages, can help
melanoma cells to survive and bypass MAP-kinase pathway targeted drug
action by producing the soluble factor TNFα. Moreover, when patients are
receiving treatment with the novel targeted drugs, macrophages produce
more TNFα, which confers resistance and makes treatment less effective.
Combining standard treatment with drugs that block TNFα action at the same
timecouldpotentially providemore long-lasting andeffective treatments to
increase survival.
One of the reasons why melanoma is such an aggressive cancer is its early
tendency to invade surrounding tissue. It is therefore important to understand
whatcontrolsmelanomainvasion.Usingazebrafishxenograftmodelallowed
us to make the striking discovery that in a heterogeneous setting, inherently
invasive cells co-invade with subpopulations of poorly invasive cells, a
phenomenon termed ‘co-operative invasion’. During co-operative invasion, the
invasive cells provide protease activity and deposit extracellular matrix (ECM).
Moreover, thepoorly invasivecellsnotonly ‘benefit’ fromtheheterogeneous
situation, but also alter the mode that the invasive cells use for invasion.
Ourdataidentifiedasofarneglectedpropertyofmelanoma,whichistheability
of melanoma cell subpopulations to cooperate within a heterogeneous tumour,
and this drives melanoma progression while at the same time preserving the
heterogeneity seen throughout tumour progression. We are currently assessing
the consequences of co-operativity in a broader sense and are investigating
how co-operative behaviour impacts on the response of melanoma cells to
MAP-kinase pathway targeted therapy.
Heterogeneity in Melanoma
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 426
Manchester Cancer Research Centre Research Report
Translating Cancer Biology into Novel Therapeutics in the Manchester Cancer Research Centre
The Drug Discovery Unit at the Manchester Cancer Research Centre (MCRC)
was established in 2009 to translate the fundamental biological research
conducted within the MCRC into novel therapeutic opportunities. Recent
advances have seen two key projects make substantial progress toward
clinical evaluation.
In late 2009, scientists at the Cancer Research UK Manchester Institute began
to unravel the biological role of a little-studied enzyme known as poly-(ADP-
ribosyl) glycohydrolase, or PARG. This enzyme functions in concert with the
PARP family of enzymes to alert cells to DNA single strand breaks, before
these lesions lead to genetic instability and damage. In recent years, the poly
(ADP ribose) polymerase (PARP) family of enzymes have been the subject of a
number of clinical trials, evaluating agents such as Olaparib from AstraZeneca
and Rucaparib from Clovis Oncology. Indeed, imminent European approval
of such agents highlights their clear therapeutic benefit in patients with
ovarian cancer.
DNA single strand breaks (SSBs) are the most common type of damage that
arises in cells. Upon damage, PARP binds to the SSB and auto-ribosylates itself
using NAD+ as a substrate. This generates chains of poly ADP ribose (PAR),
which provide a recruitment signal for repair complexes to accumulate at the
damage site and repair the lesion. Removal of the PAR chains, an essential step
to facilitiate access of this repair machinery, is accomplished by PARG, the only
enzyme known to efficiently catalyse the hydrolysis ofO-glycosidic linkages
of ADP-ribose polymers and thereby reversing the effects of PARPs. Total
PARGdeficiencyleadstocelldeathwhilstPARGdepletion,usingRNAi,leadsto
pleiotropiceffectssuchasPARchainpersistence,progressionofsingle-strand
to double-strand DNA lesions and NAD+ depletion (Figure 1). Ultimately, these
effectsshouldleadtocelldeathintumours,whereconsiderableDNAdamage
exists and alternate DNA repair pathways may be compromised.
As a therapeutic target, PARG has been largely unexplored to date. Suggestions
that the enzyme is ‘undruggable’ and scant literature information dissecting
its biological function have held back this area of research. However, work
Allan Jordan, Ian Waddell and Donald Ogilvie
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 4 27
within the Instituteand furtherafieldhassuggested thatPARGmayofferan
alternate therapeutic strategy to the more widely-investigated PARP inhibitors
in certain tumour types with increased genetic instability and compromised
DNA repair pathways.
TheDDUhasbeenactivelyinvestigatingthisnoveltargetsince2010butefforts
were initially hampered by difficulties in finding novel chemical compounds
which could form the basis of a drug discovery programme. In 2011 discussions
with AstraZeneca revealed that the company also had an interest in the target
and had potential startpoints which may be available to the DDU. Having
conducted a high throughput screen of 1.7M compounds, the AstraZeneca
teamhadidentifiedjustasingle‘hit’compound,highlightinghowcomplexthis
target was to prosecute. Crucially, the high-resolution crystal structure of this
compoundboundtohumanPARGhasbeenobtained,detailingspecificallyhow
the compound and PARG interacted on the atomic level. This information was
transferred to the DDU and work on the project began in earnest.
The team soon revealed that this early molecule could inhibit the action of
PARGinacellularcontextandwebelievethatthiswasthefirsttimethatrobust
cellular anti-PARG activity had been demonstrated. However, the compound
was also cytotoxic and suffered from poor physicochemical properties
that would preclude further development. Through expert computational
and medicinal chemistry design, strategies were enacted to ‘scaffold-hop’
away from this early hit into novel chemical matter and this approach yielded
several new chemical series for the group to investigate in a series of detailed
biological protocols.
Over the past two years, these chemical series have been refined and
optimised to deliver over a thousand-fold improvement in cellular potency
compared to the original hit and have delivered a much more drug-
like overall profile. These derivatives ablate the clonogenic potential of
certain tumour cell lines and we believe that we now understand at least
some of the factors which sensitise particular cell lines to PARG inhibition.
This information will be of critical importance in determining which
Translating Cancer Biology into Novel Therapeutics in the Manchester Cancer Research Centre
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 428
Manchester Cancer Research Centre Research Report
patients may respond best to these agents and will influence the design of clinical trials once we have
identified suitable candidatemolecules. Moreover, these compounds are well tolerated in vivo and show
promising pharmacokinetics which now allow the therapeutic role of PARG inhibition to be more
comprehensively investigated.
Critical in vivo studies are now underway within the Unit. Having demonstrated that our compounds can
engage with, and inhibit, PARG in tumours in vivo, leading to persistence of PAR chains, we are now determining
how this translates to inhibition of tumour growth in vivo.Theseexperimentswillhelpdefineourpathtoward
the clinical evaluation of these exciting and unique inhibitors.
WhilstPARGoffersanopportunitytounderstandandexploitnovelbiology,oureffortsontheREToncogenic
kinase recognises a therapeutic need for improved agents against a drug target which is already clinically
validated. The RET (Re-arranged during transfection) receptor tyrosine kinase is a known oncogene
which is implicated in certain subsets of medullary thyroid cancer (MTC) and 1-2% of non-small cell lung
cancers (NSCLC).
Whilst agents such as Vandetanib and Cabozantanib have recently been approved for use as RET inhibitors
inMTC,theiruseisnotwithoutcomplication.NeitheragentwasdesignedasaspecificRETinhibitorandthis
serendipitoussecondarypharmacologyistemperedbyconsiderableside-effectscausedbyinhibitionofother
kinases. These additional pharmacologies are dose-limiting, reducing the therapeutic utility of the agents in
the chronic clinical setting and have led to FDA ‘black box’ usage warnings for both compounds. We believe that
amoreselectiveRETinhibitorwouldbeofconsiderablebenefittopatients,allowingstrongertargetinhibition
and,therefore,increasedclinicalutilitywithreducedsideeffects.
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 4 29
Translating Cancer Biology into Novel Therapeutics in the Manchester Cancer Research Centre
Theteamhasspentconsiderabletimeandeffortdevelopingadetailedbiologicaltestingcascadethatallows
ustohighlightanysuperiorefficacyofourexperimentalagentsoverthosealreadyemployedclinically.Given
that the field of kinase inhibitors is heavily exploited in drug discovery, we have also developed innovative
approaches to reveal novel startpoints which we have now shown to be potent and selective inhibitors of RET at
both the enzyme and cell level. These diverse chemical derivatives show a range of physicochemical properties
and we are investigating their pharmacokinetics in more detail in order to prioritise those most likely to be
appropriate for further development. Our primary objective at the present time is to deliver molecules with
improvedpharmacokinetics,whilstmaintainingtheexcellentpotencyandselectivityprofileswehaveobtained.
These tool compounds will then facilitate early proof of concept experiments to demonstrate unequivocally
thatspecificandselectiveinhibitionoftheREToncogenewilldelivertherapeuticbenefitinRET-driventumour
xenograft models.
Resistance to targeted kinase inhibition is often observed through mutation of the enzyme, leading to
insensitivity of the tumour to the drug. This anticipated mechanism of resistance often arises through
alteration of a key area of the protein known as the ‘gatekeeper region’ of the kinase. Through computational
andbioinformaticsanalysis,webelievethatwehaveidentifiedthelikelyclinicalresistancemechanismand,to
circumvent the clinical implications of this issue, we have already begun the hunt for inhibitors of the resistant
enzyme. Having a second-line therapeutic approach in place in readiness for emergent resistant disease
should prolong the time over which we can control RET-driven disease in patients with NSCLC and, in doing so,
prolong overall survival.
To accelerate these studies, in November 2014 the Cancer Research Technology Pioneer Fund and Sixth
Element Capital announced that it had agreed to fund this project through to Phase I clinical trials. The fund,
a joint venture between Cancer Research Technology and the European Investment Fund, was established in
2012tohelpfillthefundinggapbetweenearlystageresearchandearlyclinicalevaluation.Thisfundingisthe
fourth investment made by the fund and provides us with access to increased capabilities and resources for
the project, in order to deliver a candidate molecule for pre-clinical evaluation before the end of 2016.
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 430
Manchester Cancer Research Centre Research Report
Further delivery of novel therapeutics will depend heavily on identifying emerging new therapeutic targets from
across the MCRC and a key activity in the team is to provide chemical tool compounds to help understand
emerging biology. Through the application of medicinal chemistry, we work closely with group leaders from
across the University to help identify, source and synthesise molecules which are not commercially available
but which may help to better understand certain biological processes. Through such interactions, we have
recently helped Cathy Tournier’s group uncover more details about the role of the ERK5 kinase in epithelial
inflammation and the role thismay play in tumourigenesis, and provided Janni Peterson’s teamwith tools
to dissect the regulation of the mitotic regulator Wee1 by TOR signalling. Ultimately, these advances in the
understandingoftumourigenesisandcellcyclecontrolmayleadtotheidentificationofnewdrugtargetswhich
can be progressed by the team. More dramatically, our collaborative work with Tim Somervaille’s group on
LSD1 has not only helped his team deliver a more comprehensive understanding of the role of this exciting
epigenetic target in acute myeloid leukaemia (AML) but has highlighted the MCRC as a key centre for the
investigation of this disease. We were delighted that this expertise was recently recognised by the decision by
Oryzon Genomics to bring their Phase I clinical trial of irreversible LSD1 inhibitors to The Christie, with Tim as
leadinvestigatoronthetrial.WefeelthisembodiesthebenefitsofcollaborativeresearchacrosstheMCRC,
deliveringearlyaccesstoemerging,first-in-classtreatmentstopatientsintheNorthWestandwehopethisis
thefirstofmanysuchtrialsweareabletohelp,insomeway,todeliverintoTheChristie.
Moving forwards, we expect that this will not be our only involvement in bringing new clinical trials to Manchester.
Over the coming months we will be further enhancing our internal target validation capabilities, increasing
oureffortsalongsideMCRCgroup leaders tostrengthentheclinical linkage foremerging targetsandbuild
confidenceintheirpotentialtractabilityasnoveldrugdiscoveryprogrammes.Thiswillallowustomaintainan
activeanddynamicportfolio,ensuringaflowofqualitytargetsintotheteam.AsourinvestmentsinPARGand
RETmovetheseprojectstowardclinicaldevelopment,thisinfluxofnewertargetswillbeimportanttomaintain
our delivery ambitions.
With the increased funding for RET now available to us, we anticipate that the next 18 months will be critical for
the project and our ambition is to accelerate the pace of our research to deliver novel candidate molecules for
pre-clinical evaluation at the end of this timeframe. Along similar lines, we anticipate that over the next year we
will secure a development partner for PARG. This partnership will draw in additional resources and greatly assist
inoureffortstonarrowdownourinitialpatientpopulationforclinicalevaluation.Thesewiderexperimentswill
thenbeusedtodefineourpaththroughpre-clinicalevaluationand,ultimately,ourearlyclinicaltrialformat.
The past two years have seen considerable progression of our internal and collaborative research portfolio;
our ambition over the next two years is to translate these dramatic advances into small molecules with the
potential for evaluation in a Phase I patient cohort.
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 4 31
Translating Cancer Biology into Novel Therapeutics in the Manchester Cancer Research Centre
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 432
Manchester Cancer Research Centre Research Report
Collaboration pays dividends for leukaemia research
Touched upon in the report from the Drug Discovery Unit was their collaboration
with Leukaemia Biology on LSD1 inhibitors – this success embodies the bench-
to-bedside approach of the MCRC.
In 2010 William Harris, a graduate student in the Leukaemia Biology group,
performed a small-scale screen of candidate epigenetic regulators in leukaemia.
ThesehadbeenidentifiedinamicroarrayexperimentwehadpublishedinCell Stem Cell in 2009 as expressed highly in murine Mixed Lineage Leukaemia
stemcells(MLLLSC)anddownregulatedwiththeirdifferentiationandlossof
stem cell potential. Bill noticed that knockdown of one of these genes, LSD1,
led toLSCdifferentiation. LSD1 is a histonedemethylasewhich is known to
remove methylation marks from histone tails, potentially thereby regulating
gene expression. He searched the literature for compounds known to inhibit
LSD1 and identified themonoamine oxidase inhibitor tranylcypromine. This
drug has been in routine clinical practice for several decades in the treatment of
depression and irreversibly inactivates LSD1 with an IC50of5-20μM.Remarkably,
whenappliedtoMLLLSCitpromotedcelldifferentiationinasimilarmannerto
LSD1 knockdown.
To identify compound classes that might serve a useful role in the clinic, we then
initiated a collaboration with the Cancer Research UK Manchester Institute’s
DrugDiscoveryUnit.AllanJordanandJamesHitchinidentifiedarecentlyfiled
patent reporting substantially more potent and selective inhibitors of LSD1.
These inhibitors are chemicallymodified versions of tranylcypromine, and it
is thechemicalmodificationof tranylcyprominewhichconfers theenhanced
potency and selectivity. James Hitchin synthesised one of these compounds
for us and we were able to demonstrate that, in the nanomolar range, we were
abletopromoteLSCdifferentiationin in vitro and in vivo assays using murine
and human cell lines and also patient cells.
Our work, which was published in Cancer Cell in 2012, has led to a fruitful
collaboration with Oryzon Genomics, based in Barcelona, Spain. It was this
companythathadfiledthepatentforthenewermorepotentLSD1inhibitorswe
were able to synthesise for our experiments. Our work enabled Oryzon to push
Tim Somervaille
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 4 33
ahead with a clinical trial focus for their advanced lead compound ORY1001,
giventheselectivesignalforLSD1inhibitorefficacyinleukaemiaversusother
sub-types of malignancy. It further enabled Oryzon to acquire orphan drug
status for ORY1001 for the treatment of acute myeloid leukaemia from the
European Medicines Agency in 2013, a designation designed to facilitate the
development of new medications for rare diseases.
The phase I trial of ORY1001 opened in July 2014 at the Hospital Universitari Vall
d’Hebron in Barcelona and The Christie NHS Foundation Trust in Manchester,
withTimSomervailleastheUKChiefInvestigator.Thisisafirst-in-class,first-
into-man early phase trial of LSD1 inhibition in acute leukaemia, with a particular
focus on uncovering the pharmacokinetic and pharmacodynamic properties
of this completely new medication. Patients receive cycles of oral therapy as
in-patients, and later as out-patients, and their disease parameters are very
carefully monitored. Recruitment is proceeding well and preliminary results of
the trial should become available during the course of 2015.
Collaboration pays dividends for leukaemia research
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 434
Manchester Cancer Research Centre Research Report
Ovarian cancer and angiogenesis
MCRC researchers have a particular interest in understanding angiogenesis in
ovarian cancer.
Angiogenesis has emerged as a novel target for anti-cancer therapies
through randomised clinical trials that have tested the benefit of adding
vascular endothelial growth factor (VEGF) inhibitors, such as bevacizumab, to
conventional cytotoxic therapies. Despite improvements in progression-free
survival,thebenefitinoverallsurvivalhasbeenmodest.Tumourangiogenesis
is regulated by a number of angiogenic cytokines. Thus innate or acquired
resistance to VEGF inhibitors can be caused, at least in part, through expression
of other angiogenic cytokines, including fibroblast growth factor 2 (FGF2),
interleukin 8 (IL-8) and stromal-cell-derived factor 1α (SDF-1α), which make
tumours insensitive to VEGF signalling pathway inhibition.
The activity of most angiogenesis-related growth factors is regulated by
heparan sulphate (HS), which is essential for the formation of FGF2/FGF
receptor (FGFR) and VEGF(165)/VEGF receptor signaling complexes. HS is a
component of cell surface and extracellular matrix proteoglycans that regulates
numerous signaling pathways by binding and activating multiple growth factors
and chemokines. The structural characteristics of HS that determine activation
or inhibition of such signaling complexes are only partially defined, but the
amount and pattern of HS sulphation are key determinants for the assembly of
the trimolecular, HS-growth factor-receptor, signalling complex.
The Translational Angiogenesis group, led by Professor Gordon Jayson,
showed that ovarian tumour endothelium displays high levels of HS sequences
that harbour glucosamine 6-O-sulphates when compared with normal
ovarian vasculature. Reduced HS 6-O-sulphotransferase 1 (HS6ST-1) or
6-O-sulphotransferase 2 (HS6ST-2) expression in endothelial cells impacted
upon the prevalence of HS 6-O-sulphate moieties in HS sequences, which
consist of repeating short, highly sulphated S domains interspersed by
transitionalN-acetylated/N-sulphateddomains.Wesawthata≤40%reduction
in 6-O-sulphates significantly compromised FGF2- and VEGF(165)-induced
endothelial cell sprouting and tube formation in vitro and FGF2-dependent
Gordon Jayson
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 4 35
angiogenesis in vivo. Moreover, HS on wild-type neighbouring endothelial
or smooth muscle cells failed to restore endothelial cell sprouting and tube
formation. The affinity of FGF2 for HS with reduced 6-O-sulphation was
preserved, although FGFR1 activation was inhibited, correlating with reduced
receptor internalisation. We concluded that 6-O-sulphate moieties in
endothelial HS are of major importance in regulating FGF2- and VEGF(165)-
dependent endothelial cell functions in vitro and in vivo and highlighted HS6ST-1
and HS6ST-2 as potential targets of novel antiangiogenic agents.
We then demonstrated that HS6ST-1 and HS6ST-2 play a direct role in
ovarian cancer angiogenesis. Down-regulation of HS6ST-1 or HS6ST-2 in
human ovarian cancer cell lines resulted in 30-50% reduction in glucosamine
6-O-sulphate levels in HS, thus impairing HB-EGF-dependent EGFR signaling
and diminishing FGF2, IL-6, and IL-8 mRNA and protein levels in cancer cells.
These cancer cell-related changes again reduced endothelial cell signaling
and tubule formation in vitro. In vivo, the development of subcutaneous
tumournoduleswithreduced6-O-sulphationwassignificantlydelayedatthe
initial stages of tumour establishment with further reduction in angiogenesis
occurring throughout tumour growth. Our results showed that in addition
to the critical role that 6-O-sulphate moieties play in angiogenic cytokine
activation, HS 6-O-sulphation level, determined by the expression of HS6ST
isoforms in ovarian cancer cells, is a major regulator of angiogenic program in
ovarian cancer cells impacting HB-EGF signaling and subsequent expression of
angiogenic cytokines by cancer cells.
Ovarian cancer and angiogenesis
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 436
Manchester Cancer Research Centre Research Report
Targeting multiple angiogenic cytokines with HS mimetics may represent an opportunity to inhibit tumour
angiogenesismoreefficiently.Ourpublishedstudiesandunpublishedworkhavedemonstratedthefeasibility
of generating synthetic HS fragments of defined structure with biological activity against a number of
angiogenic cytokines.
Working in conjunction with the Clinical and Experimental Pharmacology group, the Translational Angiogenesis
group has also been exploring potential predictive biomarkers for existing anti-angiogenic therapies. The
modest improvement in survival seen in trials of such agents has sparked interest in developing measures that
couldallowclinicianstoselectwhichpatientsaremostlikelytobenefit,whileminimisingtoxicity.Weanalysed
patient blood samples from the international ICON7 trial, which investigated the addition of bevacizumab to
conventional cytotoxic therapy in ovarian cancer.
Using multiplex ELISAs previously developed and validated to Good Clinical Practice standards, we determined
the pre-treatment plasma concentrations of 15 angiogenesis–related factors implicated in VEGF biology
(VEGFA,-C,and-D;andVEGFreceptors,VEGFR1,andVEGFR2),angiogenicfactorsinovariancancer(fibroblast
growth factor, FGF2; interleukin, IL8; angiopoietin, Ang1 and Ang2; and Tunica internal endothelial cell kinase
2, Tie2), or potential mediators of resistance to VEGF (placental growth factor, PlGF; FGF2; platelet-derived
growth factor, PDGFbb; granulocyte colony–stimulating factor, GCSF; or hepatocyte growth factor, HGF) and
investigated their predictive significance.We fitted aCoxmodel of PFS for each distinct combinationof a
putativebiomarkerandtreatmentandusedaKaplan-MeierestimatortovisualisethePFSforeachidentified
putative biomarker.
Two striking observations emerged: (i) for Ang1, as a linear model on the nontransformed continuous scale,
therewasaclearsuggestionofaninteraction,and(ii)forTie2,althoughtherewasnosignificantinteraction
term, there was clear evidence of heterogeneity in the relation between high levels of Tie 2 and treatment. For
women with high Ang1/low Tie2 values, treatment with bevacizumab had an expected HR of 0.21 for PFS when
comparedwithstandardtreatment.ThebiologicimplicationofthesefindingisthattheAng1–Tie2axismay
play a pivotal role in mediating resistance to VEGF pathway inhibitors.
The teamhas identified that the angiopoietin signalling systemprovides predictive information that could
optimise the use of VEGF pathway inhibitors in the treatment of ovarian cancer. Further as yet unpublished
data have shown that the angiopoietin family of molecules also provides information on the development of
resistancetoVEGFinhibitors.Whileconfirmatorystudiesareplanned,iftheresultsarevalidatedtheworkwill
allow the investigators to identify whom to treat with VEGF inhibitors and alert the treating team to the onset
of vascular resistance to these drugs so that new agents can be instituted.
Inthe laboratory,theteamhasdeterminedthestructuresofsyntheticoligosaccharidesthat inhibitdefined
angiogeniccytokines;providing thefirstevidence forstructural specificity in thisfieldand therebyopening
the door to a new family of saccharide-based anti-angiogenic therapeutics that would address the resistance
identifiedthroughthebiomarkerprogrammedescribedabove.
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 4 37
Ovarian cancer and angiogenesis
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 438
Manchester Cancer Research Centre Research Report
Key role for Radiotherapy in personalised approach to cancer treatment
Highprofilerecruitmentandconfirmedfundingfornewcutting-edgefacilities
combine with a strong research output to result in a successful two years for
Radiotherapy-Related Research at the Manchester Cancer Research Centre.
International expertise in physics has been strengthened by two Chair-level
appointments. Professor Marcel van Herk, a world-renown radiotherapy
physicist currently based at the Netherlands Cancer Institute in Amsterdam, will
move to Manchester in April 2015. Professor van Herk has carried out pioneering
workinthefieldofimaginginradiotherapyandearlyinhiscareerdevelopedan
electronic portal imaging system that has since been commercialised and used
worldwide. More recent work has continued to pioneer innovative approaches
to the application of image-guided intensity-modulated radiotherapy.
A second appointment will drive forward research in proton therapy. Manchester
has recruited Professor Karen Kirkby, who has a particular interest in ion beam
therapyanditsbiologicaleffects,asChairinProtonTherapyPhysics.Professor
Kirkby was previously Director of Science for the Surrey Ion Beam Centre and
played a key role in getting proton therapy on the political agenda.
The last two years have also seen progress towards the establishment of both
proton therapy and novel image-guided radiotherapy facilities in Manchester.
FollowingconfirmationoffundingfromtheDepartmentofHealthforaProton
Therapy Centre to be located at The Christie, research groups have received
approval to include an additional gantry in order to establish a proton research
facility. Within the University’s School of Physics, Dr Hywel Owen is leading
research into accelerator technology, the development of Monte Carlo codes
and methods for particle tracking, and the design of novel storage rings for
synchrotron radiation.
Plans to replace the research radiotherapy linac in the Wade Centre at The
Christie were boosted by the announcement that Manchester had joined
Elekta’s international collaboration to develop a magnetic resonance (MR)
image-guided radiotherapy system. A prototype MR Linac has been developed
in Utrecht and now we have the opportunity to contribute to the clinical
implementation of this cutting-edge technology.
Tim Illidge, Catharine West and Nick Slevin
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 4 39
The Translational Radiobiology group investigates potential prognostic and
predictive biomarkers, and in particular is interested in assessing radiosensitivity,
predicting late radiation toxicity effects and exploring the utility of hypoxia-
modifying agents. Two collaborative publications have made use of several
sets of radiogenomic study data: RAPPER (Radiogenomics: Assessment of
Polymorphisms for Predicting the Effects of Radiotherapy), RADIOGEN and
Gene-PARE(GeneticPredictorsofAdverseRadiotherapyEffects).Agenome-
wide association study in prostate cancer patients, in conjunction with groups in
Santiago de Compostela, Spain, University of Cambridge and Mount Sinai New
York,identifiedasusceptibilitylocusforlateradiotherapytoxicityat2q24.1.This
locus comprises TANC1, which is known to play a role in regenerating damaged
muscle. A larger study, exploring association of common SNPs with toxicity two
years after radiotherapy in 1850 breast and prostate cancer patients, provided
evidence of a relationship. 2014 also saw the start of the REQUITE project, which
aims to validate predictive models of radiotherapy toxicity alongside biomarkers
of radiosensitivity. The international study will collect patient-reported outcome
and quality of life data from 5,300 radiotherapy patients. Such a patient-centred
approach to assessing toxicity has been demonstrated by the Radiotherapy in
LungCancergrouptobeeffectiveinalungcancerpatientcohort.
Duetothekeyroleplayedbyoxygenationindefiningradiosensitivity,Professor
Catharine West’s group has also been developing gene-based hypoxia
signatures and investigating the potential of such signatures and other hypoxia
measures to predict benefit from hypoxia-modifying therapy. In a cohort of
high-riskbladdercancerpatients,necrosiswasshowntopredictbenefitfrom
hypoxia modification. Within the same cohort of patients from the BCON
Key role for Radiotherapy in personalised approach to cancer treatment
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 440
Manchester Cancer Research Centre Research Report
(bladder carbogen nicotinamide) trial, we found that those with high HIF-1αexpressiongainedbenefitfromthe
additionofcarbogenandnicotinamidetostandardradiotherapy,whereasnobenefitwasseenwithlowHIF-1α.
A study looking at the BCON cohort alongside laryngeal cancer patients enrolled on the ARCON (accelerated
radiotherapy with carbogen and nicotinamide) trial demonstrated that a 26-gene hypoxia signature predicted
benefitfromthehypoxiamodifyingtherapyinlaryngealbutnotbladdercancer.
Within the Targeted Therapy and Experimental Oncology groups, there is a shared interest in using novel
immunomodulatory agents to potentiate the effect of ionising radiation. In 2014,we reported pre-clinical
experiments using a systemically administered Toll-like receptor 7 agonist and showed that administration
of this agent in combination with ionising radiation primes an antitumour CD8+ T-cell response leading
to improved survival in colorectal carcinoma and fibrosarcoma models. In addition, efficacy extended
outside the irradiationfield, reducingmetastatic load.Asecondstudyevaluatedanapproach toovercome
acquired resistance to fractionated radiotherapy. We demonstrated that radiotherapy leads to an adaptive
upregulation of PD-L1 expression in tumour cells that is dependent on CD8+ T-cell production of IFNγ and
mayattenuate theefficacyof the anticancer immune response.By deliveringmAb targeted againstPD-1
andPD-L1,wewereabletoenhancetheefficacyoflowdosesoffractionatedradiotherapy.Dosescheduling
for the anti PD-L1 agent was critical, with effective anti-tumour effects only seen with concurrent not
sequential administration.
Professor Tim Illidge’s group are also involved in trials of radioimmunotherapy agents. At the beginning of 2014,
resultswere reported froman internationalmulti-centrephase II trial evaluating theefficacy andpotential
toxicity of fractionated 90Y-ibritumomab tiuxetan (90Y-IT) as an initial therapy for follicular lymphoma. The study
demonstrated that fractionated radioimmunotherapy using this agent was effective and well-tolerated in
patients with advanced stage disease.
Investigators at The Christie have led several trials looking at methods to improve outcome in head and neck
cancer.NIMRAD is a phase III trial investigating the use of hypoxiamodification in the formof nimorazole
alongsideIMRT,withaparticularaimtoassessthebenefit inpatientswhoarenotsuitableforsynchronous
chemotherapyorcetuximab.Itishopedthatthisstudywillleadtomoreeffectivetreatmentoptionsforsuch
patients,whoaretypicallyelderlyorinfirm.
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 4 41
Key role for Radiotherapy in personalised approach to cancer treatment
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 442
Manchester Cancer Research Centre Research Report
Imaging Science research at the MCRC
Advanced imaging methods play a core role in cancer research, particularly in
thefieldsofradiotherapyrelatedresearch,drugdevelopmentandpersonalised
medicine. Developing and translating validated imaging biomarkers is a
significantchallenge,butonethatiscentraltotheaimsoftheMCRC.
Imaging research within the MCRC has been bolstered substantially by nearly
£9M funding from Cancer Research UK (CRUK) and the Engineering and
Physical Sciences Research Council (EPSRC) to establish a joint cancer imaging
centre between The University of Manchester and the University of Cambridge.
Thisawardbegan inDecember2013andwill run forfiveyears.Thestrategic
importance of the funding was summed up by Dr Iain Foulkes, Cancer Research
UK’s executive director of strategy and research funding, who said: “Imaging is
aninvaluabletoolinthefightagainstcancer.Beingabletoseewhat’shappening
inside a patient is vitally important in understanding how treatments are working
and the best ways to improve them”.
As part of the CRUK-EPSRC Cancer Imaging Centre, scientists within the
MCRC undertake world-class research using a variety of techniques, such as
bioluminescence and optical microscopy, MRI (Magnetic Resonance Imaging)
and PET (Positron Emission Tomography). This latest funding has helped to
recruit PhD students, clinical fellows and post-doctoral researchers who are
helping a number of PIs to develop new imaging techniques and applications.
Our portfolio covers a range of projects centred on tumour angiogenesis,
hypoxia, cell signalling and death that link into the three research themes
identifiedpreviouslyintheMCRC,namely(1)thedevelopmentandvalidationof
imaging biomarkers; (2) development of preclinical imaging and (3) translational
imaging research. Particular focus is made on two cancer types – lung and brain
–thatareidentifiedbyCRUKascancersofunmetneed.
Imagingbiomarkervalidationandqualification
All biomarkersmust be validated and then qualified for clinical use. Imaging
biomarkers are no exception and the series of steps required to validate
an imaging biomarker differ from biospecimen biomarkers. CRUK and the
European Organisation for Research and Treatment of Cancer (EORTC) have
recognisedthesekeydifferencesandinvestigators inManchesterare leading
JamesO’Connor,AdamMcMahon,GeoffParker,KayeWilliamsand Alan Jackson
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 4 43
Imaging Science research at the MCRC
aninternationalconsensusefforttoproducearoadmapforimagingbiomarker
studies in oncology. This work has been led by CRUK Clinician Scientist Dr
James O’Connor, Professor John Waterton and Professor Alan Jackson and
has over 75 collaborators from major European and North American institutions
including the NIH/NCI.
The philosophy espoused in this initiative has potential to transform the way in
which imaging biomarker science is carried out internationally. The initial output
has already been adopted by CRUK as criteria for assessing imaging biomarker
applications and a landmark paper will be published in early 2016 in Nature Reviews Clinical Oncology. This has already helped to consolidate the MCRC’s
position as a world leader in both biospecimen and imaging biomarker science.
Tissue microstructure evaluation with MRI
Imaging tissue microstructure in vivo may provide a clinical tool for detecting
tumour cell proliferation and death and tumour infiltration into neighbouring
healthy tissues. Tissue microstructure can be assessed using diffusion
magneticresonanceimaging(dMRI)whichissensitivetothediffusionoftissue
water over distances of a few microns. However, clinical use of dMRI has two
significantlimitations:Firstly,itisimpossibletorelatechangesintheapparent
diffusioncoefficient(ADC)ofwatertochangesincellsize,packingdensityor
membranepermeabilityunambiguouslytocancercell infiltration,proliferation
and therapy response. Secondly, current strategies for reducing the corruption
of dMRI data due to motion are unsatisfactory and can blur tumour features or
substantially extend scan durations.
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 444
Manchester Cancer Research Centre Research Report
Work ledbyProfessorGeoffParkerhasdemonstrated thatdMRIcanquantify tissuemicrostructure in the
brain and other neural tissues, by measuring axon diameter in the whole brain. This principle has been extended
to measure tumour cell packing, density and cell membrane permeability. Further work has used an analytical
expression to create an interactive tool for exploring the signal changes arising for tissues with different
properties. This has shown that sensitivity to change in intracellular volume fraction is lower for larger cells than
smaller cells, and that the sensitivity varies with imaging sequence parameters. It has also allowed a comparison
of the signal changes predicted when cells shrink but density either changes or remains stable. This novel work
highlightsthecompetingeffectsofchangesincellsizeandvolumefractionondiffusionsignals(Figure1).
Figure 1: Interactive tool for evaluating tissue cell size and density Screenshotofthetoolshowssignalchanges(colours)asafunctionofpulsesequenceparameters(G,Δ,∂)andtissueproperties(R,Di,De,fi).Images courtesy of Dr Damien McHugh and Dr Fenglei Zhou.
These biomarkers are being validated using a number of strategies including novel tissue mimetic phantoms,
comparing MRI measurements with scanning electron microscopy and with micro-CT data performed at the
Henry Moseley X-ray Imaging Facility in Manchester. Once validated, the dMRI measurements will be tested in
clinical cohorts against other MRI biomarkers of the tissue microstructure including 23Na MRI, in collaboration
with Dr Ferdia Gallagher and colleagues in Cambridge.
ImagingoxidativedamageandinflammationwithnovelPETtracers
Positron emission tomography is an exquisitely sensitive imaging modality capable of yielding both images and
quantitative data regarding tracer uptake kinetics, distribution and excretion, using sub-microgram quantities
of tracer. Dr Adam McMahon is leading work to combine this sensitivity with the selectivity of antibodies
towardsspecificbiomolecules,whereantibodiesarechemically radiolabelledwithaPET isotope.Theteam
at the Wolfson Molecular Imaging Centre (WMIC) have already demonstrated ability to perform such labelling
experiments binding 89Zr to the two monoclonal antibodies: cetuximab and bevacizumab. 89Zr is used because
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 4 45
Imaging Science research at the MCRC
Figure 2: Mass spectroscopy imaging of tumour lipid metabolism A-H&EimageshowingtwoNSCLCtumours.B:MSimagesofthesametissue,showingthetumourspecificdistributionof a lipid species.
its half-life (3.27 days) allows the antibody to be imaged over the 2 – 3 day period required for the antibody to
reach its target. Future work will aim to improve this approach through the use of nanobodies (small fragments
of antibodies that retain the desired property of selective target recognition but due to their small size can
reach their targets within a few hours). Radiolabelling methods that chemically bond these nanobodies to the
PET isotope 18F have been developed, which give better quality PET images than 89Zr with less exposure to
radiation. Thefirstnanobodytobelabelledtargetsthemacrophagemannosereceptorallowingtheroleof
activatedmacrophagestobeimagedintumour-associatedinflammatoryprocesses.
To image oxidative damage, PET tracers are under development that will bind to radical sites in tissues
generated by exposure to ionising radiation (as in radiotherapy) or oxidative chemical insults (as experienced in
some types of chemotherapy).
Lipid Metabolism in Tumours
Tumourlipidmetabolismdiffersfromlipidmetabolismseeninhealthytissues.BothPETandmassspectrometry
(MS) imaging methods are being developed to image these differences, led by Dr Adam McMahon and
Professor Kaye Williams. PET powerfully allows metabolic processes to be imaged in real time but is sensitive
to only one substance (the PET tracer) at a time. To allow PET imaging of lipid metabolism we have radiolabelled
a fatty analogue ([18F]FTHA). MS imaging requires sections of surgically removed tissue but can potentially
image hundreds if not thousands of biomolecules or pharmaceuticals in that tissue section.
ThisisillustratedintheimagesinFigure2.EachMSimageshowsadifferentlipidcomponentofthetumours
anditisclearlyseenhowthetumoursandhealthysurroundingtissuesdifferintherelativeconcentrations
of the three lipids imaged. The non small cell lung cancer (NSCLC) tumour bearing tissues were supplied by
Dr David Lewis and Professor Kevin Brindle, our collaborative partners at The University of Cambridge.
MR Imaging of tumour hypoxia
Hypoxia is critical in promoting genomic instability in tumours and determining therapeutic response, so
can drive the complex genetic heterogeneity observed in all human solid cancers. Mapping tumour oxygen
tensionandhypoxiacanhavesignificantimpactonpatientmanagementbethatthroughselectingappropriate
treatments or monitoring response. Current hypoxia assays involve biopsy and histological sampling, but these
are invasive and do not provide comprehensive tissue coverage. Imaging approaches are attractive in that
they are non-invasive, enable multiple measurements and allow whole tumour mapping. Oxygen-enhanced
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 446
magnetic resonance imaging (OE-MRI) is a promising technique for quantifying tissue oxygenation status and
tumour hypoxia, which has been developed as a cancer biomarker in Manchester.
Work led by Professor Kaye Williams and Dr James O’Connor is focussing on the biological validation of
quantitative OE-MRI measurements in multiple tumour models, including 786-O (Figure 3), SW620, U87 and
Calu6 xenografts. Initial studies have shown that OE-MRI measurements detect chronic hypoxia and when
combined with dynamic contrast enhanced MRI (DCE-MRI) can track evolution of tumour hypoxia and response
to hypoxia targeted agents and radiotherapy. Part of this work has been in collaboration with Dr Simon Robinson
at the Institute of Cancer Research. Ongoing work is looking at the relationship of MRI measurements to PET
tracers of hypoxia (18F-FAZA) and photoacoustic methods of imaging hypoxia, the latter in collaboration with
Dr Sarah Bohndiek in Cambridge. Evaluation of therapy induced responses and test-retest reproducibility has
begun in clinical studies in patients with rectal cancer and NSCLC.
Mapping and quantifying tumour heterogeneity
Most tumours demonstrate biological heterogeneity with variation in genomic features and local environmental
factors such as hypoxia, which can stimulate local genetic mutation and the evolution of treatment resistant
tumour cells. Regional variations in proliferation, cell death, metabolic activity, vascular structure and other
factorsarecommon. Imagingoffersawide rangeof tools toassessbothclonalandcellularheterogeneity
but extensive development and optimisation is required. Although imaging biomarkers of heterogeneity
can be associated with disease progression, therapeutic response and malignancy, information concerning
heterogeneity is rarely utilised and there is no consensus concerning the choice of descriptive metrics.
Manchester has developed heterogeneity metrics that clearly outperform conventional summary metrics in a
number of areas involving diagnosis, tumour grading and treatment response. Current work, led by Professor
Alan Jackson, focuses on developing the potential of imaging-based heterogeneity measures using both MRI
and PET tracers. Initial studies to statistically combine data are focussing on existing glioblastoma data sets with
anatomical,diffusionweightedanddynamiccontrastMRI.Theapproachwillthenbeextendedtoencompass
PETdata,evaluatedintissuesaffectedbyphysiologicalmotion.Neuro-oncologicalMRdatawillenableusto
validate and test our methods. Next, studies will integrate serial clinical imaging and biopsies (including liquid
biopsies) with the aim of detecting and unravelling response/resistance induced changes to both novel agents
and repurposed drugs. Immunohistochemical and genetic analysis - whole genome sequencing, whole exome
Manchester Cancer Research Centre Research Report
Figure 3: Combined OE-MRI and DCE-MRI tumour segmentation and relationship to pathology In panel a, OE-MRI and DCE-MRI data from a renal cancer 786-O xenograft are combined. Blue voxels have a ‘hypoxia’ signature and map spatially to the tumour region shown to be hypoxic on IHC staining of pimonidazole adduct formation (panel b). Yellow voxels (in panel a) map spatially to well perfused tumour regions as revealed by Hoechst 33342 staining.
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 4 47
Imaging Science research at the MCRC
sequencingandRNAsequencing-ofbiopsieswillbeusedtoqualifyimagingbiomarkerbasedclassifications,
in collaboration with Professor Richard Marais. The multi-dimensional data (genomic, automated microscopy-
image analysis and radiological images) will be integrated by developing a robust integrated database and
analysis pipeline with proper visualisation tools to give a systems view of cancer at diagnosis and as it evolves
in response to treatment.
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 448
Manchester Cancer Research Centre Research Report
Biobank building on success
The Manchester Cancer Research Centre Biobank was established in 2007
with the ultimate aim of making quality tissue collection and retrieval easier
for the researcher. There are a number of elements which are vital in building a
successful Biobank and the MCRC Biobank has approached each one carefully
to ensure that a robust infrastructure and service is provided to stakeholders
who wish to use biological samples for high quality cancer research.
The four key considerations when building a successful Biobank (Figure
1) include; compliance with the regulatory framework, which includes The
Human Tissue Act and Ethics Approval, ensuring adequate logistics for timely
procurement of specimens of a variety of types, quality sample collection by
following standard operating procedures and a robust informatics infrastructure
to ensure samples can be linked to clinical data and can be tracked and
traced appropriately.
Jane Rogan
Informatics Quality
RegulatoryFramework
Logistics
Figure 1: Key Elements of a Successful Biobank
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 4 49
The MCRC Biobank community consists of the core work team, which is
established under The Christie Division of Research, the Histology Core Facility
within the Cancer Research UK Manchester Institute and a cross-cutting team
ofclinicalstaff,suchassurgeons,pathologistsandoncologists,withoutwhom
the Biobank would not be able to function.
Since inception, the Biobank has proven its success in supporting research
within the MCRC and beyond. Samples have been collected from over 6500
patients and the Biobank has approved almost 100 Biobank applications to
use collected samples. To serve the Manchester cancer research community
as a whole, samples are collected across many solid tumour types. Figure
2 demonstrates the spread of solid tumour sample collection over the life
of the Biobank.
In addition to the solid tumour bank, the MCRC Biobank also has a haematological
malignancy arm, which collects blood and bone marrow from patients with
leukaemia and other blood disorders (see Figure 3). Although separately run
and managed through the CRUK MI histology core facility, it falls under the
MCRC Biobank research tissue bank ethics approval and access to samples is
facilitatedthroughaunifiedaccesspolicyforbothsolidandliquidtumours.
Evolutionofaflexiblemodel
In the seven years since the MCRC Biobank was established, it has responded
to research requirements by developing a flexible model to ensure that
researchers’ needs can be met. Originally set up with the premise that a six-
packof fixed and frozen tumour, normal andmatchedblood andurine from
patients undergoing surgery would be banked for future use, it soon became
Biobank building on success
Figure 2: Solid tumours Figure 3: Liquid tumours
Collection By Disease Group - Solid Tumours
Brain Breast Skin Colorectal
Upper GI Gynae Urology Lung Other
Collection By Disease Type - Haematological Malignancy
AML B-ALL MDS Other
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 450
Manchester Cancer Research Centre Research Report
apparent that this would only satisfy a proportion of research need. The Biobank was approached to facilitate
sample collection beyond this initial aim and it also became important to ensure that collected samples and
sample types were useful and being used.
To facilitate bespoke requests, the Biobank has had to respond rapidly to manage the logistical and ethical
challengesthiscanbring.Thefirstsignificantchangewastheshift frompurely retrospectivebiobankingto
more prospective sample collection, driven by both the requirement for samples which can only be collected
prospectivelyandtheneedtoensurefiniteresourceisusedasefficientlyaspossible.
Specific requests for samplesnow facilitated through theBiobank includecollectionof fresh tissue,which
requirescoordinationofstafftoensuresamplescanbedeliveredimmediatelytotheresearcherandthequality
of the sample can be maintained. This is often coupled with the need for consent to be taken to allow tissue
to be used for the creation of animal models of disease, a requirement under The Human Tissue Act. This
requires detailed planning on both the Biobank and research side where tumour size, type of media used and
cold ischaemia time all impact on the viability of the sample for its intended use.
To further meet research requirements, an update to the Biobank’s research tissue bank ethics approval now
allows a single consent to enable serial collection of samples, something which has facilitated research studies
involving the collection of blood from patients throughout the course of their treatment. The new forms also
allow the patient to be consented for collection of alternative, non-invasive sample types, such as ascites,
pleuralfluidandpluckedhair.
For each tailored requirement, the Biobank has to consider various elements which may impact the collection
and each condition can bring several different considerations. Table 1 demonstrates the various flexible
requirements that the MCRC Biobank now facilitates and the various considerations for each.
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 4 51
Biobank building on success
Requirement Considerations
Prospective Fresh Tissue• Availability of tissue• Staffavailableforsampledelivery• Media for keeping samples viable
Use for Animal Models• Specificconsentforanimalmodels• Size of tumour tissue
AlternativeSampleTypes(fluids,pluckedhair) • Routes / logistics of sample collection
Serial Blood Collection (including CTCs)• Biobank Technician presence in clinic • Oncologists willing to take patient consent
Past samples (e.g. pre-treatment/primary tumours)• Specificconsent• Requesting blocks from local hospitals
Samplingtumourindifferentareas • Further input from pathology
Clinical Trial Sample Collection• Space for sample storage• Differentprotocolsforeachtrial
Inadditiontocreatingaflexiblemodelforsamplecollection,theBiobankalsoappliesthesameethostodata
collection. This has been facilitated by obtaining a permission called Section 251 support from the Health
Research Authority, which allows linkage of large archival tissue sample cohorts, falling outside of the consent
provisions of The Human Tissue Act, to be married to clinical datasets. This enables follow-up and outcome
data to be linked to the tissue samples so that research looking at markers for survival and disease progression
can be carried out.
Added Value
In addition to its core work, in recent years the Biobank has also acted as a vehicle to deliver a number of
nationalresearchinitiatives,whichdemonstratetheeffectivenessoftheBiobanktodeliverlarge-scalesample
collection within the Manchester Cancer Research Centre.
Thisbegan inJuly2011withPhase1oftheCancerResearchUKStratifiedMedicineProgramme(SMP1),a
pilot study to demonstrate on a small scale how the NHS can provide molecular diagnosis for all cancer types
routinely. The MCRC Biobank was selected to act as one of seven Clinical Hubs to collect samples and data
frompatientsacrossfourtumourtypes.SectionsfromeachsamplecollectedweresenttoCardiffTechnical
Hubwhereaspecificpanelofuptosixgenespertumourtypewastested.Manchesterwasaconsistentlyhigh
recruitertoSMP1andwastheonlyClinicalHubtosignificantlyoverachieve;collectingsamplesfromnearly
1000 patients in less than two years.
TheBiobankisnowinvolvedinPhase2oftheStratifiedMedicineProgramme(SMP2),whichhasalungfocus,
and the Biobank are working closely with MCRC lung cancer researchers to deliver this. SMP2 will feed directly
into the National Lung Matrix trial which will be opening in 2015 and this will allow patients to access targeted
treatments specifically linked toanygenetic aberrations identifiedaspartof thegeneticpanel carriedout
through the programme.
Due to Manchester’s success in delivering SMP1, Cancer Research UK selected the Biobank to act as a centre
for the 100,000 Genomes pilot, a precursor to the Genomics England (GEL) 100,000 Genomes Project, a
government funded initiative aiming to sequence the genome of 100,000 NHS patients and their families in
cancer and rare diseases.
Table 1: Facilitating sample collection for a range of requirements
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 452
Manchester Cancer Research Centre Research Report
Being part of the pilot has enabled a strong working relationship to develop between the MCRC Biobank and
the Manchester Regional Genetics Service at St Mary’s Hospital, who have been responsible for the DNA
extraction of blood and tissue collected from patients and who have been running a parallel rare diseases pilot.
At the end of 2014, Central Manchester University Hospitals NHS Foundation Trust (CMFT) and The Christie
NHS Foundation Trust submitted a joint bid to become a Genomics Medicine Centre (GMC) for the 100,000
Genomes Project and were successfully selected in the first wave. CMFT will act as the overall lead for
Manchester and for rare diseases, with The Christie leading on the cancer element. It is anticipated that the
cancer sample collection will begin in May 2015.
In addition to the 100,000 Genomes Project and SMP, the Biobank is also facilitating sample collection
for a number of lung specific studies, including TRACERx, a mesothelioma bank called MesoBank and is
collaborating with the Manchester Respiratory and Allergy Biobank (MANRAB) to collect normal lung tissue for
non-cancer lung-related research.
Future Developments
The growth of the MCRC and the new MCRC building present a real opportunity for the Biobank to also grow and
develop further to meet the needs of researchers. 2015 will see the development of a new, more streamlined
AccessPolicy tocopemore readilywith thegrowingdemand for samplesand further staff recruitment to
ensure that the MCRC’s research objectives are not hindered by lack of resource for tissue collection in key
disease areas.
The Biobank will continue to work hard to its mission statement:
Facilitating high quality cancer research by bringing a flexible and committed approach to ethical sample and
data collection
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 4 53
Biobank building on success
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 454
Manchester Cancer Research Centre Research Report
Author Biographies
Caroline Dive is a Senior Group Leader at the CRUK Manchester Institute and
Professor of Pharmacology at The University of Manchester. After completing
her PhD studies in Cambridge, Caroline moved to Aston University’s School
of Pharmaceutical Sciences in Birmingham where she started her own group
studying mechanisms of drug induced tumour cell death. She then moved to
what became the Faculty of Life Sciences at The University of Manchester to
continue this research. Caroline was awarded a Lister Institute of Preventative
Medicine Research Fellowship before moving to the CRUK Manchester Institute
in 2003. Here she set up the Clinical and Experimental Pharmacology group
interfacing with the early clinical trials unit at The Christie.
Andrew Hughes is Chair in Cancer Experimental Therapeutics and Strategic
Director of the Experimental Therapeutics Unit at The Christie NHS Foundation
Trust. He graduated with a double first in Medical Sciences at Cambridge,
spending 3 years as a tutor in Physiology and Bye-fellow of Downing College,
Cambridge, whilst completing a PhD in Behavioural Neuroendocrinology
in the Department of Anatomy. Andrew subsequently practised General
Clinical Medicine in Manchester’s teaching hospitals in the UK, before joining
AstraZeneca in 1994. Until 2015 he held a dual appointment as Professor of
Translational Medicine at The University of Manchester and Head of Early
Clinical Oncology Development at AstraZeneca.
Jonathan Tugwood is Senior Translational Scientist in the Clinical and
Experimental Pharmacology (CEP) group, CRUK Manchester Institute.
Following a BSc in Biological Sciences at Birmingham University and a PhD in
Molecular Virology at Manchester University, he undertook post-doctoral
studies in embryonal genetics at Columbia University, New York. This was
followed by a post-doctoral position in Molecular Toxicology with ICI (now
AstraZeneca) at the Alderley Park site, which became a permanent team leader
role in 1991. Jonathan moved to AstraZeneca Pharmaceuticals in 1997 to join
the Global Safety Assessment function as a team leader, carrying out original
research in drug toxicology, and providing support for drug discovery and
development programmes. He joined CEP in 2012, and has responsibility for
theCEPbiomarkertrialsportfolio, includingoversightofthestaffdeveloping
new circulating biomarker assays and delivering biomarker data for CEP’s trials
and experimental medicine portfolio.
Andrew Hughes, Jonathan Tugwood, Caroline Dive, Matt Krebs, Emma Dean - Ambitious plans for Experimental Therapeutics - Page 8
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 4 55
Emma Dean is a clinical senior lecturer within the Institute of Cancer Sciences
at The University of Manchester and Honorary Consultant in Medical
OncologyatTheChristieNHSFoundationTrust.Shequalified inMedicineat
The University of Nottingham, graduating with a Bachelor degree in Medical
Sciences, Bachelor of Medicine and Bachelor of Surgery. After completing
general medical training, she took up a specialist post in Medical Oncology at
The Christie, Manchester. She was awarded a PhD in 2009 for research into
novel anti-cancer therapies that promote cancer cell death through a Cancer
Research UK Fellowship. Postgraduate research as a National Institute for
Health Research Clinical Lecturer focussed on early phase clinical trials using
novel targeted small molecule inhibitors, next generation chemotherapeutics,
and anti-angiogenics, either alone or in combination with licensed therapies
or radiotherapy. Her current research interest is first-in-human therapies in
medicaloncologywithaspecificinterestinmoleculartargetedapproaches.
Matthew Krebs is a clinical senior lecturer within the Institute of Cancer
Sciences at The University of Manchester. He completed his degree in Medicine
at The University of Leicester in 2001 and took opportunity to spend an
undergraduate elective period at the London Regional Cancer Centre, Ontario,
Canada. Following general medical training in Manchester he commenced
specialist training in Medical Oncology at The Christie NHS Foundation Trust in
2005. Matthew subsequently joined the Cancer Research UK/AstraZeneca PhD
Clinical Fellowship programme in November 2007 and was a clinical research
fellow within the Clinical and Experimental Pharmacology group at the Cancer
Research UK Manchester Institute. His research focuses on the isolation and
characterisation of circulating tumour cells from patients with lung cancer with
a view to developing these as predictive and/or pharmacodynamic biomarkers
in early phase clinical trials.
Tony Howell is Professor of Medical Oncology at The University of Manchester
and, along with Gareth Evans and Nigel Bundred, he leads the Nightingale Centre
and Genesis Prevention Centre at University Hospital of South Manchester
NHS Foundation Trust. He is the former Director of the Breakthrough Breast
Cancer Research Unit, and the Manchester Breast Centre. He trained at
Charing Cross Hospital, London and, after a period with the Medical Research
Council, moved to Birmingham and then took a post to lead Breast Medical
Oncology at The Christie NHS Foundation Trust in Manchester. Formerly
he was Chairman of the UK Breast Trials Organisation (UKCCCR), the British
Breast Group and the ATAC trial and, up until the end of 2007, was the Research
&DevelopmentDirectorofTheChristieNHSFoundationTrustandResearch
DirectoroftheGreaterManchester&CheshireCancerResearchNetwork.His
interests are the endocrine therapy and biology of the breast and breast cancer
with a particular interest in prevention. He has published over 480 papers mainly
in these areas.
Author Biographies
Tony Howell, Rob Clarke, Gareth Evans - Breast Cancer research at the Manchester Breast Centre - Page 14
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 456
Rob Clarke carried out undergraduate BSc studies in Biology at the University
of Sussex and the Université de Grenoble in France, graduating in 1987.
Following two and half years as a Research Assistant with Professor Potten at
the Paterson Institute for Cancer Research, his PhD studies investigated the
control of proliferation in the normal and neoplastic human mammary gland.
Subsequent post-doctoral training with Dr Liz Anderson was in the Clinical
Research Department of The Christie, Manchester. In 2001, he returned to The
University of Manchester as a Cancer Research UK Research Fellow, becoming
a Group Leader. Currently, he is Senior Lecturer and Deputy Director of the
Breakthrough Breast Cancer Research Unit in the University of Manchester’s
Institute of Cancer Sciences based at the Paterson Building.
D Gareth Evans has a national and international reputation in clinical and
research aspects of cancer genetics, particularly in neurofibromatosis and
breast cancer, and is Chairman of the NICE (National Institute for Clinical
Excellence) Familial Breast Cancer Guideline Development Group. He has
developed a clinical service for cancer genetics in the North West Region and
lectures within the UK and internationally on hereditary breast cancer and
cancer syndromes. He has also developed a regional training programme
for clinicians, nurses and genetic associates in breast cancer genetics, and
established a system for risk assessment and counselling for breast cancer in
Calman breast units.
Fiona Blackhall trained in Manchester and for two years in Toronto with Dr
Frances Shepherd. She was appointed Consultant Medical Oncologist at The
Christie NHS Foundation Trust in 2005, joining the Manchester Lung Group led
by Professor Nick Thatcher. She was awarded an Honorary Senior Lecturer role
at The University of Manchester in 2007. She is an active clinical trialist with a
focus on development of mechanism based therapies and has a major role in
biomarker research. She is Chair of the translational subgroup of the recently
established European Thoracic Oncology Platform, a member of the NCRI
Lung Clinical Studies Group and leads the MCRC lung cancer research team.
Manchester Cancer Research Centre Research Report
Caroline Dive, Fiona Blackhall, Phil Crosbie, Rajesh Shah - Circulating tumour cell (CTC) research in early stage, resectable non-small cell lung cancer (NSCLC) - Page 18
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 4 57
Phil Crosbie is a Consultant Respiratory Physician based at the University
Hospital of South Manchester with a special interest in lung cancer. He started
his training in Edinburgh before moving to Manchester where he completed
a PhD exploring the molecular epidemiology of lung cancer at The University
of Manchester, prior to taking up an NIHR Clinical Lectureship. He delivers
interventional bronchoscopy services both at UHSM and The Christie. His
current research interests include the early detection/staging of lung cancer
and the study of circulating biomarkers and biology of circulating tumour cells
in early stage disease. His laboratory work is based in Professor Caroline Dive’s
CEP group at the CRUK Manchester Institute.
Claudia Wellbrock is a Reader in the Faculty of Life Sciences at The University
of Manchester. She received an MSc in Chemistry from the University of
Wuerzburg, Germany and during her PhD project at the Max Planck Institute
of Biochemistry in Wuerzburg studied the oncogenic function of receptor
tyrosine kinases. She continued with postdoctoral studies at the University of
Wuerzburg focusing on the signal transduction pathways involved in pigment-
cell transformation and melanoma development. In 2002, she joined the
laboratory of Richard Marais at the Institute of Cancer Research in London, UK,
where she investigated the role of BRAF in the initiation and development of
melanoma. She moved to The University of Manchester in September 2007
and her current work aims to understand the crosstalk between particular
cellular signalling cascades in melanoma initiation and progression in order to
identify new targets for cancer therapy.
Donald Ogilvie heads the Drug Discovery Unit at the MCRC. He joined the
Cancer Research UK Manchester Institute as a senior group leader in February
2009 after a twenty year career in the pharmaceutical industry. Donald obtained
anMAinBiochemistryatOxfordin1980beforeworkingattheJohnRadcliffe
Hospital for eight years on the role of proteases in breast cancer then inherited
connective tissue disorders. The latter was the basis of his D.Phil degree. In
1988 he joined ICI, which subsequently became Zeneca then AstraZeneca.
For most of his industrial career, Donald worked on cancer drug discovery and
early clinical development and he was directly responsible for the delivery of ten
novel cancer development compounds, several of which have progressed to
PhaseII&IIIclinicaltrials.
Author Biographies
Claudia Wellbrock - Heterogeneity in Melanoma - Page 24
Allan Jordan, Ian Waddell and Donald Ogilvie - Translating Cancer Biology into Novel Therapeutics in the Manchester Cancer Research Centre - Page 26
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 458
Allan Jordan joined the Drug Discovery Unit in July 2009 as Head of Chemistry.
After gaining a BSc in Chemistry from UMIST in 1993 and a short spell as a
teaching assistant in Arizona, he returned to UMIST to conduct post-graduate
research into anticancer natural products. After post-doctoral work at the
University of Reading, he joined RiboTargets in Cambridge (now Vernalis) where
he worked on a number of therapeutic areas at various stages of the research
pipeline. Alongside involvement in a number of oncology programmes,
ultimately leading to the clinical evaluation of Hsp90 inhibitors in conjunction
with Novartis, he became involved in CNS research programmes where he was
a project leader on a GPCR drug discovery programme and was also involved in
the management of Vernalis’ clinical programme for Parkinson’s disease.
Ian Waddell joined the Drug Discovery Unit in June 2011 as Head of Biology.
He gained a BSc and PhD in Biochemistry at the University of Dundee. After a
short spell as a post-doctoral research associate, he spent 5 years as a lecturer
in Molecular Medicine in the Department of Child Health at Ninewells Hospital,
before joining Zeneca in 1994. His interest in Oncology began when he led the
Cachexia team looking at preventing the skeletal muscle wasting associated
with pancreatic cancer.
Tim Somervaille trained in Medicine at St Mary’s Hospital Medical School (now
part of Imperial College London) and University College London. Following
postgraduate training in General Medicine in London, he underwent specialist
haematology training at UCL where he also studied for a PhD as a Medical
Research Council Clinical Training Fellow in Professor Asim Khwaja’s laboratory.
He then spent four years undertaking postdoctoral studies in leukaemia in
Michael Cleary’s laboratory at Stanford University as a Leukaemia Research
Fund Senior Clinical Fellow. He now leads the Leukaemia Biology group at the
Cancer Research UK Manchester Institute and is also Honorary Consultant in
Haematology at The Christie NHS Foundation Trust.
Manchester Cancer Research Centre Research Report
Tim Somervaille - Collaboration pays dividends for leukaemia research - Page 32
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 4 59
Gordon Jayson is Professor of Medical Oncology at The University of
Manchester and leads the glycoangiogenesis research within the Institute of
CancerSciences.Hequalified inMedicineat theUniversityofOxfordbefore
undertaking medical and oncology training in Manchester at The Christie.
Following a PhD in heparan sulfate biology, Gordon conducts research that
aims to translate new data and understanding of heparan sulfate biology gained
from fundamental research into the clinic.
Tim Illidge is Professor of Targeted Therapy and Oncology at The University
of Manchester. He is also Honorary Consultant in Oncology at The Christie,
with a clinical interest in the management of lymphoma. Tim completed his
undergraduate degree at London University, before studying medicine at
Guy’s Hospital Medical School and then completing a PhD at the University
of Southampton. Following this, he completed research fellowships in
Southampton and at Stanford University in California. Currently Tim is chair
of the National Cancer Research Institute (NCRI) Clinical and Translational
Radiotherapy (CTRad) group and serves within national and international
Lymphoma groups.
Nick Slevin is Director of Networked Services at The Christie and Honorary
ProfessorofClinicalOncology atTheUniversityofManchester.Hequalified
in medicine from Birmingham in 1978 and completed postgraduate training
in General Medicine in South Warwickshire and New Zealand. Nick trained
in Oncology in Nottingham and Manchester and has specialised in the non-
surgicalmanagementofheadandneckcancer since1988.Hewas thefirst
Chair of the National Cancer Research Institute Head and Neck Group and the
RoyalCollegeofRadiologists’firstleadoftheheadandneckelectronicnetwork.
Author Biographies
Gordon Jayson - Ovarian cancer and angiogenesis - Page 34
Tim Illidge, Catharine West and Nick Slevin - Key role for Radiotherapy in personalised approach to cancer treatment- Page 38
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 460
Catharine West leads the Translational Radiobiology group at The University
of Manchester. She studied Biology as an undergraduate at York University and
Radiobiology as a postgraduate at the Institute of Cancer Research, Sutton.
After postdoctoral work at the University of Rochester Cancer Centre, New
York, she joined the Paterson Institute for Cancer Research in 1986, and in
2002, joined The University of Manchester.
James O’Connor read Medicine at Magdalene College, University of
Cambridge and at the Royal Free Hospital, London, graduating with an MA in
History and Philosophy of Science and with the MB, BS degrees. After MRCS,
he began specialist training on the Manchester Radiology Training Scheme
in September 2004. He joined the Imaging Sciences department at The
University of Manchester in 2005 initially studying for a PhD, developing MR
and CT imaging biomarkers of tumour microvascular heterogeneity. This was
funded by a Cancer Research UK Clinical Research Training Fellowship. Post-
doctoral research was as an NIHR and Wellcome Trust funded Clinical Lecturer
in Radiology, from 2010. He was appointed Senior Lecturer with an Honorary
Consultant Radiologist post at The Christie in January 2012 and the following
year began a four-year intermediate Fellowship with a Cancer Research UK
Clinician Scientist Award.
Adam McMahon studied at The University of Oxford, The University of Bristol
and Dalhousie University in Canada, where he gained a BA (Hons) in Chemistry,
an MSc in Analytical Chemistry and a PhD in Chemistry. In 1988 he became
Harwell-Wolfson Research Fellow, based at the UKAEA’s Harwell Laboratory and
Wolfson College, Oxford. Then in 1991 he became Manager for Research and
Development in the AEA Technology’s Analytical Sciences Centre at Harwell.
Before joining the Wolfson Molecular Imaging Centre in January 2005, he
worked for 12 years as a Senior Lecturer in Chemistry and Analytical Chemistry
Section Leader at Manchester Metropolitan University.
Manchester Cancer Research Centre Research Report
James O’Connor, Adam McMahon, Geoff Parker, Kaye Williams and Alan Jackson - Imaging Science research at the MCRC - Page 42
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 4 61
Geoff Parker leads the QBI Lab and the Unit of Neuroimaging and Tractography
(UNIT) at The University of Manchester. He is also Director of a University of
Manchester spin out company, BiOxyDyn Limited.
Alan Jackson joined The University of Manchester in 1993 and was appointed
Professor of Neuroradiology in the Imaging Sciences Group within the Faculty
of Human and Medical Sciences at The University of Manchester in 1995. Prior
to this, he was a full-time NHS clinical neuroradiologist at the Manchester Royal
Infirmary. Alan holds honorary chairs inNeuroimaging at theUniversities of
Liverpool and Bangor and is also an honorary clinical consultant at the Salford
HospitalsNHSTrustandtheManchesterRoyalInfirmary.
Kaye Williams received a BSc in Molecular Biology from The University of
Manchester and a PhD from the Paterson Institute for Cancer Research.
Following Research Associate and Research Fellow positions within the
Experimental Oncology Group of the School of Pharmacy and Pharmaceutical
Sciences, headed by Professor Ian Stratford, she was awarded the British
Association for Cancer Research/AstraZeneca Frank Rose Young Scientist
Award in 2005. In January 2006 Kaye was appointed Senior Lecturer within
the School of Pharmacy, became Reader in 2010, and in 2012 was promoted
to Chair in Experimental Therapeutics and Imaging. She currently heads the
Hypoxia and Therapeutics group and is the MCRC lead for Preclinical Imaging.
Author Biographies
M a n c h e s t e r C a n c e r R e s e a r c h C e n t r e R e s e a r c h R e p o r t 2 0 1 3 / 1 462
Manchester Cancer Research Centre Research Report
Jane Rogan is Business Manager of the MCRC Biobank and has a wider role at
The Christie NHS Foundation Trust as HTA Designated Individual for research.
Jane has extensive experience of human tissue research and its accompanying
regulation and has been involved in tissue banking and its related governance
for ten years. Jane has a Biological Sciences undergraduate degree from The
UniversityofBirminghamandanMBAfromHuddersfieldUniversityBusiness
School. She is currently studying for an MSc in Healthcare Leadership as part of
the NHS Leadership Academy Elizabeth Garrett Anderson Programme.
Noel Clarke is a Consultant Urological Surgeon at The Christie and Salford
Royal Hospitals in Manchester and Professor of Urological Oncology at The
UniversityofManchester. HequalifiedinmedicineatCharingCrossHospital,
London, in 1981 and gained full accreditation as a Urological Surgeon in 1993.
Noel is Chairman of the National Cancer Research Institute (NCRI) Prostate
Clinical Studies Group, Chairs the European Organisation for Research and
Treatment of Cancer (EORTC) Prostate Disease Orientated Group and is Chair
of the Greater Manchester and Cheshire Urology Network Clinical Studies
Group. He directs the Genito-Urinary (GU) research group and is the Principal
Investigator of the Manchester arm of the Medical Research Council (MRC)
Northern Prostate Cancer Collaborative. Noel’s research has focused on
cancer stem cell biology and the pathophysiology of metastatic behaviour, with
translational studies directed towards analysis of new biomarkers, advanced
cancer imaging and the evaluation of novel therapies.
Jane Rogan - Biobank building on success - Page 48
Manchester Cancer Research CentreThe University of Manchester
Wilmslow Road
Manchester
M20 4BX
tel: +44 (0) 161 446 3156
fax: +44 (0) 161 446 3109
www.manchester.ac.uk/mcrc