PhD Study : Development of gene-editing technologies to aid diagnosis of type 1 diabetes

Summary

Type 1 diabetes (T1D) is an autoimmune disease characterized by apoptosis of insulin secreting pancreatic beta-cells. As a result, people with T1D are unable to produce insulin in response to physiological demands and treatment therefore focuses on insulin replacement strategies. T1D accounts for ~10% of all cases of diabetes equating to a global prevalence of approximately 45M people. TNFAIP3 is a negative regulator of inflammation and apoptosis and protects against beta-cell apoptosis in experimental models. Variants in TNFAIP3 are associated with type 1 diabetes (T1D) risk and poor residual beta-cell function. Furthermore, TNFAIP3 protects against beta-cell apoptosis, the primary driver of T1D development. It is unclear whether the association between variants in TNFAIP3 and T1D risk/beta-cell function result from a direct effect on beta-cell survival, whether these variants create a heightened inflammatory environment, which indirectly impacts on the beta-cell, or whether both mechanisms may have a role to play in T1D development.

AIM: The aim of this pilot study is to determine how variants in TNFAIP3 affect beta-cell function and survival and whether these variants have diagnostic and/or prognostic value for T1D.

OBJECTIVES:

1. To recruit a cohort of people with T1D from the Western Health and Social Care Trust (WHSCT) and to study the prevalence of variants in TNFAIP3 and their association with age of onset, glycaemic control (HbA1c) and residual beta cell function (C-peptide). Recruitment protocols are already in place and we have established good working relationships with clinical staff in the WHSCT. Findings from this study will be validated in data from secondary cohorts from Exeter University.  The top three ranked variants will be carried forward for further analysis (objectives 2 and 3).

2.To conduct an assessment of the direct effect of variants on the beta-cell, the top three ranked variants in TNFAIP3 will be introduced into the MIN6 beta cell line using CRISPR/Cas9 technologies. The effect on cell survival (TUNEL assay, PCR for markers of apoptosis) and function (glucose-induced insulin secretion) will be determined.

3. To conduct an assessment of the indirect effects of these variants on the beta-cell, inflammatory cells (CD4+ T cells and monocytes) isolated from individuals carrying each of the top three ranked variants will be isolated from blood samples collected during construction of our cohort (objective 1). Following stimulation (PMA activation of monocytes to macrophages; cytokine challenge of T cells) of these cells, the conditioned media will be added to MIN6 cells and primary islets, and the effects on cell survival and function assessed (as outlined in Objective 2). Many of the TNFAIP3 variants associated with T1D risk are common variants (>20% of the population) located in intergenic regions. Therefore, it is difficult to predict function. However, this is an important question to address since these variants have been reported in multiple independent studies as being predictive/associated with T1D development. To turn these variants into useful clinical biomarkers, we must understand function. Via gene-editing technologies, this project will determine the diagnostic/prognostic value of these variants in T1D.

Essential criteria

Applicants should hold, or expect to obtain, a First or Upper Second Class Honours Degree in a subject relevant to the proposed area of study.

We may also consider applications from those who hold equivalent qualifications, for example, a Lower Second Class Honours Degree plus a Master’s Degree with Distinction.

In exceptional circumstances, the University may consider a portfolio of evidence from applicants who have appropriate professional experience which is equivalent to the learning outcomes of an Honours degree in lieu of academic qualifications.

• Sound understanding of subject area as evidenced by a comprehensive research proposal
• A comprehensive and articulate personal statement

Desirable Criteria

If the University receives a large number of applicants for the project, the following desirable criteria may be applied to shortlist applicants for interview.

• First Class Honours (1st) Degree
• Masters at 65%
• Completion of Masters at a level equivalent to commendation or distinction at Ulster
• Research project completion within taught Masters degree or MRES
• Experience using research methods or other approaches relevant to the subject domain
• Work experience relevant to the proposed project
• Publications - peer-reviewed
• Experience of presentation of research findings

Funding and eligibility

The University offers the following levels of support:

Vice Chancellors Research Studentship (VCRS)

The following scholarship options are available to applicants worldwide:

• Full Award: (full-time tuition fees + £19,000 (tbc))
• Part Award: (full-time tuition fees + £9,500)
• Fees Only Award: (full-time tuition fees)

These scholarships will cover full-time PhD tuition fees for three years (subject to satisfactory academic performance) and will provide a £900 per annum research training support grant (RTSG) to help support the PhD researcher.

Applicants who already hold a doctoral degree or who have been registered on a programme of research leading to the award of a doctoral degree on a full-time basis for more than one year (or part-time equivalent) are NOT eligible to apply for an award.

Please note: you will automatically be entered into the competition for the Full Award, unless you state otherwise in your application.

Department for the Economy (DFE)

The scholarship will cover tuition fees at the Home rate and a maintenance allowance of £19,000 (tbc) per annum for three years (subject to satisfactory academic performance).

This scholarship also comes with £900 per annum for three years as a research training support grant (RTSG) allocation to help support the PhD researcher.

• Candidates with pre-settled or settled status under the EU Settlement Scheme, who also satisfy a three year residency requirement in the UK prior to the start of the course for which a Studentship is held MAY receive a Studentship covering fees and maintenance.
• Republic of Ireland (ROI) nationals who satisfy three years’ residency in the UK prior to the start of the course MAY receive a Studentship covering fees and maintenance (ROI nationals don’t need to have pre-settled or settled status under the EU Settlement Scheme to qualify).
• Other non-ROI EU applicants are ‘International’ are not eligible for this source of funding.
• Applicants who already hold a doctoral degree or who have been registered on a programme of research leading to the award of a doctoral degree on a full-time basis for more than one year (or part-time equivalent) are NOT eligible to apply for an award.

Due consideration should be given to financing your studies. Further information on cost of living