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PhD Studentship in the Leeds Institute of Cancer and Pathology
Faculty of Medicine and Health
University of Leeds
ORGANOID AND SINGLE CELL MODELS OF BLADDER CANCER
Supervisors: Professor Maggie Knowles and Professor Steve Evans
A full time NC3Rs-funded PhD studentship is available. We welcome applications from both
UK and EU rated applicants, as defined in the Research Council UK criteria on residence
eligibility.
The studentship will attract an annual tax-free stipend at Research Council rate (currently
£14,296 per annum) for up to 3 years, subject to satisfactory progress and will cover the
UK/EU tuition fees.
You should have or expect to obtain a minimum of an upper second class degree in either a
biological science, physics or biophysics, a strong motivation to develop multidisciplinary
skills relevant to translational research and a commitment to the principles of the 3Rs.
Candidate whose first language is not English must provide evidence that their English
language is sufficient to meet the specific demands of their study, the Faculty minimum
requirements are:
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British Council IELTS - score of 6.5 overall, with no element less than 6.0
TOEFL iBT - overall score of 92 with the listening and reading element no less than
21, writing element no less than 22 and the speaking element no less than 23.
This multidisciplinary project aims to develop a range of in vitro organoid models from fresh
bladder tumour tissues and use these to develop drug screening assays.
Research Project:
More than 10,000 patients in the UK and 74,000 patients in the US are diagnosed with
bladder cancer each year. There are two patient groups with distinct prognoses and
molecular profiles. Non-muscle invasive bladder cancer frequently recurs and requires longterm disease monitoring and repeated localized therapy. Muscle-invasive bladder cancer is
life-threatening and requires radical treatment. Treatments have not advanced for decades
and targeted approaches to both systemic and localized therapy are needed.
Genome sequencing and expression profiling has recently defined multiple molecular
subtypes and identified a range of potential therapeutic targets, generating optimism that
novel therapeutic approaches can now be developed. This will require both functional
investigations and preclinical drug assessment, for which relevant disease models are
essential. Preclinical drug evaluation commonly uses cell line xenografts and patient-
derived xenograft models. This project aims to develop in vitro models for evaluation that
can replace a major proportion of such animal use.
Studies of other tissue types demonstrate that normal and tumour-derived human cells can
be maintained for long periods as three-dimensional organoids that are suitable for rapid in
vitro drug screening and can be modified to provide paired isogenic lines for functional
analyses.
Cellular progenitors of the two major bladder cancer clinical subgroups have been described
and specific cell surface markers that allow their isolation have been identified and used to
flow sort these cells. It has also been reported that such cells can generate 3-dimensional
sphaeroids in culture, but culture conditions have not been optimized for long-term organoid
maintenance and banking.
Project objectives:
 Optimization of conditions for tumour cell isolation, organoid culture and long-term
maintenance.
Fresh bladder tumour tissues will be used. More than 250 new bladder cancers are
diagnosed in Leeds each year and we collect 4-5 fresh tumour samples each week with
patient consent. In addition to material for culture, a portion of each tissue is snap-frozen for
molecular characterisation and a blood sample collected to provide germline DNA. A large
panel of extensively-characterised bladder cancer cell lines is also available for optimisation
experiments.
 Establishment of a frozen bank of organoids representing the range of bladder
cancer molecular subtypes.
Parallel stored tissues from successful cultures will be characterised as part of other work
within the group. This will include targeted mutation screening using a next generation
sequencing assay and genome-wide expression profiling. This will assign samples to
previously defined molecular subtypes and will identify targetable mutations.
 Development of microfluidics platform-based drug sensitivity analysis for rapid
evaluation of patient samples.
A microfluidics platform (already available) that allows trapping of individual cells and
exposure to drug profiles will be used to determine drug sensitivity to cytotoxic and cytostatic
agents. Initial experiments will use a tumour cell line with known sensitivity to FGFR
inhibitors and to cisplatin. Surface Acoustic Wave Dielectrophoresis (SAW-DEP) will be
evaluated for separation of viable from non-viable cells, prior to the sensitivity testing. Both
flow-sorted single cell populations from tumours and cells derived from organoids will be
assessed. Recent advances in the integration of spheroid / organoid formation with
microfluidic devices offer potential advantages in terms of improved screening capability and
will be tested.
The project will use a wide range of methods, including tissue dissociation methods,
organoid culture from tissue fragments and single cells, flow sorting, cell staining
(immunohistochemistry and immunofluorescence), light and fluorescence microscopy
(including confocal) and drug sensitivity testing in 2D and 3D cultures and on the
microfluidics platform. The student will be able participate in the molecular characterisation
of the originating tumour tissues depending on their interests and/or focus more extensively
on the biophysical aspects of the project.
You will be based in the University of Leeds in the Section of Molecular Oncology, Leeds
Institute of Cancer and Pathology, University of Leeds and will also work closely with
Professor Evans group in the Faculty of Mathematics and Physical Sciences. Training will
be provided in all relevant techniques.
This is a collaborative project between the groups of Professor Maggie Knowles (Leeds
Institute of Cancer and Pathology)
http://medhealth.leeds.ac.uk/profile/900/854/margaret_anne_knowles
and Professor Steve Evans (Physics)
http://www.mnp.leeds.ac.uk/steve-evans.html
References:
Knowles, M.A. & Hurst, C.D. Molecular biology of bladder cancer: new insights into
pathogenesis and clinical diversity. Nature Reviews Cancer 15, 25-41 (2015).
Aine, M., Eriksson, P., Liedberg, F., Hoglund, M. & Sjodahl, G. On Molecular Classification
of Bladder Cancer: Out of One, Many. Eur Urol 68, 921-923 (2015).
Clevers, H. Modeling Development and Disease with Organoids. Cell 165, 1586-1597
(2016).
van de Wetering, M. et al. Prospective derivation of a living organoid biobank of colorectal
cancer patients. Cell 161, 933-945 (2015).
1Chan, K.S. et al. Identification, molecular characterization, clinical prognosis, and
therapeutic targeting of human bladder tumor-initiating cells. Proc Natl Acad Sci USA
106, 14016-14021 (2009).
Volkmer, J.P. et al. Three differentiation states risk-stratify bladder cancer into distinct
subtypes. Proc Natl Acad Sci USA 109, 2078-2083 (2012).
Kwapiszewska, K., Michalczuk, A., Rybka, M., Kwapiszewski, R. & Brzózka, Z. A
microfluidic-based platform for tumour spheroid culture, monitoring and drug
screening. Lab Chip 14, 2096–2104 (2014).
How to apply:
To apply for this scholarship applicants should complete a Faculty Scholarship Application
form and send this alongside a full academic CV, degree transcripts (or marks so far if still
studying) and degree certificates to the Faculty Graduate School [email protected]
We also require 2 academic references to support your application. Please ask your referees
to send these references on your behalf, directly to [email protected] by no later than
Monday 20 February 2017
If you have already applied for other scholarships using the Faculty Scholarship Application
form you do not need to complete this form again. Instead you should email
[email protected] to inform us you would like to be considered for this scholarship project.
Any queries regarding the application process should be directed to [email protected]
Closing date for this studentship is Monday 20 February 2017