Cell Biology/Genetics: Identifying the mechanisms controlling HPGD expression and the functional consequences of HPGD increase

Prostaglandin E2 (PGE2) is a small lipid molecule frequently elevated during inflammation and cancer through increased synthesis by cyclo-oxygenase 2 (COX-2). This promotes the progression of disease. Although NSAIDs inhibit COX and have been beneficial for treatment of a variety of conditions, they can have substantial side effects (1). Evidence suggests this is because COX is situated at the start of the prostaglandin metabolic pathway, so its inhibition impacts on the much wider lipid signalling pathway components. The HPGD gene codes for the 15-hydroxyprostaglandin dehydrogenase protein, sometimes referred to as PGDH, and acts further downstream of COX. It is the key enzyme in removal of prostaglandins, and its loss causes debilitation due to the underlying proliferative changes (2). It offers an alternative therapeutic target for the reduction in PGE2 levels found in cancer and inflammation through its increased expression, and subsequent rapid PGE2 removal. By acting downstream of COX it is less likely to likely to alter the prostaglandin and wider lipid pathways.

<p>We have generated clonal cell lines that overexpress HPGD and caused a functional change: reduced cell migration. We have generated HPGD promoter constructs and identified potential DNA sequences that control expression.&nbsp;</p> <p>The project aims to:</p> <ul> <li>investigate the mechanisms by which HPGD reduces cell migration, identify if additional genes are altered in the lipid metabolism pathways. Determine which other signalling pathways are affected through increased HPGD expression&nbsp;</li> <li>identify mechanisms to increase HPGD expression.&nbsp;</li> </ul> <p>The PhD will identify and validate the genes/proteins/DNA sequences involved in HPGD regulation and consequences of overexpression so generating an understanding of the molecular mechanisms involved.&nbsp;<br /> &nbsp;</p> <h6>Techniques associated with project</h6> <p>A range of biochemical, molecular biology, cell biology, and bioinformatic techniques will be used. Further transcriptome analysis is required to generate a high-throughput screen of the clonal cell lines to identify gene expression changes due to HPGD over-expression. This will involve RNA-Seq library production and bioinformatic analysis. qPCR and western blotting, for validation. Additional functional studies may require si-knockdown, manipulation and cloning of sequences for transient over-expression analysis. HPGD promoter reporter constructs will be transfected and activity quantified. Further validation will require mutagenesis/deletion of DNA sequences and functional studies of transcription factor activity.</p> <h6>References</h6> <ol> <li>Fanaroff AC, Roe MT. Contemporary Reflections on the Safety of Long-Term Aspirin Treatment for the Secondary Prevention of Cardiovascular Disease. Drug Saf. 2016;39(8):715-27. doi: 10.1007/s40264-016-0421-1.</li> <li>Uppal S, Diggle CP, Carr IM, Fishwick CW, Ahmed M, Ibrahim GH, Helliwell PS, Latos-Biele?ska A, Phillips SE, Markham AF, Bennett CP, Bonthron DT. Mutations in 15-hydroxyprostaglandin dehydrogenase cause primary hypertrophic osteoarthropathy. Nat Genet. 2008 Jun;40(6):789-93. doi: 10.1038/ng.153. Epub 2008 May 25. Erratum in: Nat Genet. 2008 Jul;40(7):927. PMID: 18500342.</li> </ol> <p>&nbsp;This project is part of the&nbsp;<a href="https://medicinehealth.leeds.ac.uk/leeds-institute-research-st-james/doc/international-phd-academy-medical-research">International PhD Academy: Medical Research</a></p> <p><strong>In line with the bespoke nature of our International PhD Academy a modified PhD project can be proposed dependent on students interests and background.</strong></p>

<p>Please note these are not standalone projects and applicants must apply to the PhD academy directly.</p> <p>Applications can be made at any time. You should complete an <a href="https://medicinehealth.leeds.ac.uk/faculty-graduate-school/doc/apply-2">online application form</a> and attach the following documentation to support your application.&nbsp;</p> <ul> <li>a full academic CV</li> <li>degree certificate and transcripts of marks (or marks so far if still studying)</li> <li>Evidence that you meet the programme&rsquo;s minimum English language requirements (if applicable, see requirement below)</li> <li>Evidence of funding to support your studies</li> </ul> <p>To help us identify that you are applying for this project please ensure you provide the following information on your application form;</p> <ul> <li>Select PhD in Medical Research as your programme of study</li> <li>Give the full project title and name the supervisors listed in this advert</li> </ul>

A degree in biological sciences, dentistry, medicine, midwifery, nursing, psychology or a good honours degree in a subject relevant to the research topic. A Masters degree in a relevant subject may also be required in some areas of the Faculty. For entry requirements for all other research degrees we offer, please contact us.

Applicants whose first language is not English must provide evidence that their English language is sufficient to meet the specific demands of their study. The minimum requirements for this programme in IELTS and TOEFL tests are: &bull; British Council IELTS - score of 7.0 overall, with no element less than 6.5 &bull; TOEFL iBT - overall score of 100 with the listening and reading element no less than 22, writing element no less than 23 and the speaking element no less than 24.

<p>Informal enquires about regarding the bespoke taught first year of the PhD programme and research projects can be made by contacting LIMRPhD@leeds.ac.uk.</p> <p>Enquiries regarding the application process should be directed to the Faculty of Medicine and Health Graduate School e: <a href="mailto:fmhpgradmissions@leeds.ac.uk">fmhpgradmissions@leeds.ac.uk</a></p>