Elucidating cellular behaviours and potential oncogenicity utilising in vitro 3D organoid culture system

Supervisors: Primary: Dr Joo-Hyeon Lee (University of Cambridge) and Dr Mick Fellows (AstraZeneca)

Assessment of cell transformation (i.e. the acquisition of malignant characteristics in morphology, growth control or function) has proven problematic to model in vitro. The soft agar cell transformation assay has been validated, but only in a limited number of immortalised or mixed population embryonic cells and the specificity of the assay has been questioned. Furthermore, the genetic basis of cell transformation in this model has not been fully delineated (Harvey et al 2015, Creton et al 2012). The use of genetically and phenotypically stable organoids derived from single human stem cells (Schwank et al 2013) should provide a better model for cell proliferation and transformation.

Organoids have the advantage of 3D morphology, being able to be derived from several major organs and can be engineered to demonstrate proliferative effects by introducing a cell type specific ki67 fluorescent protein to tag. Organoid stability is also dependent on several growth factors and inhibitors e.g. EGF, Wnt, BMP inhibitor, TGFb inhibitor. Growth following withdrawal of these factors, which are known pathways implicated in cancer genesis, will be indicative of cell phenotypic and/or transformative changes. It is also proposed that this model could be used to investigate mechanisms of pro-oncogenicity and also the potential off-target effect of genome editing, for which there is currently an unmet need for in vitro assays to assess concerns around inappropriate editing in oncogenes.

The project will develop organoids from a variety of tissue sources, including organoids with the ki67 tag, and assessment of proliferative and morphological changes following withdrawal of growth factors after treatment with reference carcinogens and following transfection with specific and promiscuous gene editing nucleases. Comparative data will be generated from alternative in vitro methodologies to analyse cell transformation e.g. the soft agar assay. Reference carcinogens will be analysed in this assay along with the potential of gene editing nucleases to induce cell transformation (which has not been previously assessed). The student will also use next generation sequencing technologies to identify specific genes involved in morphological changes in both cell and organoid cultures. Success will provide new and more relevant model for understanding and assessment of morphological changes and carcinogenesis.

Applications
How to Apply
1. Please complete our departmental Application Form process here* (http://study.stemcells.cam.ac.uk/study/otheropportunities/bbsrc-lee-fellows-application-page), which will include submitting your CV, providing the details of two referees, and your Degree transcripts.
2. Please ask your referees to submit references directly to the SCI Graduate Administrator: [Email Address Removed], using "BBSRCiCASE (Lee-Fellows) student reference" in the subject header. They must be signed and on headed paper but can be scanned and emailed to us. It is important for referees to mention 'Lee-Fellows' in the subject header or body of the email, as we have more than one BBSRC iCASE studentship at the moment, with the same closing date.
Application Deadline: Midnight UK time on Friday 31st March 2017.

Academic Enquiries to: Dr. Joo-Hyeon Lee [Email Address Removed]. Dr Lee is happy to receive informal enquiries about this studentship.
Application Process Enquiries: Graduate Administrator [Email Address Removed]. Please mention that your enquiry relates to 'BBSRC iCASE (Smith)' in the subject header, as we have more than one BBSRC iCASE studentship.
For further details about our group and the institute, please visit:http://www.stemcells.cam.ac.uk/research/pis/lee
*Please note that for this project you DO NOT APPLY DIRECTLY TO BBSRC, but apply to us at the Wellcome Trust MRC Cambridge Stem Cell Institute, as in 'How to Apply' above