Lineage tracing and imaging of brain tumour stem cells

Applications are invited for a PhD studentship at the Division of Neuropathology and Department of Neurodegenerative Disease at the UCL Institute of Neurology under the supervision of Prof. Sebastian Brandner and in collaboration with Prof Marino, Brain Tumour Research Centre of Excellence, Queen Mary University London. The anticipated start date is 1st May, 2017.

Project
To study the initiation, clonal expansion and progression of intrinsic brain tumours by lineage tracing and three-dimensional imaging.

Summary: We will use established three-dimensional tissue models for lineage tracing. This will generate fluorescently tagged brain tumour cells which will be imaged and analysed three-dimensionally.

Brain tumour generation: brain tumour cell lines, generated from previous projects, will be used as described previously [1, 2]. Tumours generated from these cell lines are very similar to those in humans. They express classical glial and stemness markers.
Lineage tracing reveals in vivo stem-cell behaviour in its physiological context and identifies actual stemness using a bi-cistronic knock-in with fluorescent marker to allow isolation of these cell populations. In a second approach, lentiviral vectors will be generated to transduce human tumour initiating cells in vitro. The system makes use of two components: one construct will express, under the control of a stemness promoter of choice (e.g. Nestin, Gli, GFAP, Sox), the tamoxifen (TAM)-inducible CreER(T2) (in this context termed “switch”), following growth in a three-dimensional matrix, these cells will in the beginning express no label until the host receives a pulse of tamoxifen (TAM), after TAM-mediated cre expression cells are labelled in one of the four colours, GFP (green), YFP (yellow), CFP (blue), and RFP (red).

3D visualisation of cells: conventionally immunolabelled or engineered fluorescent cells are evaluated on two-dimensional histological sections. A restricted three-dimensional cellular relationship can be assessed on thick tissue sections. In order to achieve a truly three-dimensional visualisation, on deeper lying structures without mechanical disruption we will adopt a tissue clearing technology. This allows for the three-dimensional visualisation of immune-labelled or genetically engineered, fluorescent cells.
Environment

The student will be tightly integrated in our laboratory at the division of neuropathology, primarily supervised by Prof Brandner. The student will have the opportunity to attend UCL courses in relevant subject areas, as well as to interact with students and postdoctoral researchers from other departments.

Person specification

A good degree in a biology/neuroscience subject (or equivalent EU/overseas degree), but ideally an MSc in a related area.
Excellent experimental skills, preferably in cell or molecular biology
Good analytical/mathematical skills, preferably with some knowledge of bioinformatics approaches.
A strong interest in stem cell biology and cellular imaging
Good communication skills - especially in written English.
A strong work ethic, with the ability to think creatively and work independently.
Duties and Responsibilities
Develop the model system for lineage tracing. This includes setting up tissue culture conditions, transfecting cells, and
Develop the image acquisition methodology, in particular multicolour confocal microscopy
Work in collaboration with our image analyst to develop an algorithm to analyse three-dimensional relationships of colour-labelled stem and progenitor cells. A specialist software is available for this purpose
Work in collaboration with other researchers, in particular collaborators at Queen Mary University to establish and refine the tissue clearing methodology
Prepare progress reports on research as required.
Prepare manuscripts for submission to peer-reviewed journals.
Prepare presentations, including text and images, for delivery by self and others.
Travel for training, collaboration and other meetings or conferences.
Contribute to the overall activities of the research team and department as required.
Maintain an awareness and observation of Fire and Health and Safety Regulations at UCL and other facilities to be visited. Actively follow UCL policies including Equal Opportunities and Race Equality policies.

Informal enquiries
Please email Prof. Sebastian Brandner for further information about the project ([Email Address Removed]).

Application procedures
Application is by CV and covering letter (including motivation for applying) emailed to [Email Address Removed]. Candidates short-listed for interview will be required to give a short research presentation. Please put “BTR PhD Studentship” in the subject line.

Closing Date: 7th April 2017