Unravelling the Complex Pathogenic Mechanisms of RYR2 Exon-3 Deletion Using an Inducible Stem Cell Model

About the Project

Catecholaminergic polymorphic ventricular tachycardia (CPVT) is a genetic arrhythmia syndrome characterized by the onset of life-threatening arrhythmias during exertion or emotional stress. CPVT is caused by mutations of the cardiac ryanodine receptor gene (RYR2). Mutations of RYR2 predispose to the onset calcium waves delayed afterdepolarization that lead to arrhythmias especially following adrenergic stimulation. Most of these mutations are missense mutations (substitution of one amino acid). Deletion of the third exon of RYR2 (exon-3 deletion) causes a more complex syndrome characterized by a combination of typical CPVT, dilated cardiomyopathy and sinus node dysfunction. The mechanisms responsible for this complex syndrome are not understood. The main objective of this project will be to elucidate these mechanisms and identify potential treatment strategies. An inducible pluripotent stem cell (iPSC) line with exon-3 deletion will be generated from the control line using well-established genome editing techniques. The iPSC will be differentiated into cardiac ventricular myocytes and sinus node myocytes using standardized differentiation protocols. A detailed transcriptomic study will be performed to understand whether exon -3 deletion causes any alterations in the cardiac transcriptome that can explain the onset of cardiomyopathy. Fluorescent indicators (both biological and non-biological) will be utilized to characterize Calcium transient and action potential characteristics both at baseline and following adrenergic stimulation. Ca waves, delayed afterdepolarizations and arrhythmias will be induced using adrenergic stimulation and/or cardiac glycosides. Finally we will test the efficacy of various anti-arrhythmic agents in preventing calcium waves, delayed afterdepolarizations and arrhythmias.

Candidates are expected to hold (or be about to obtain) a minimum 2:1 Bachelors Degree with Honours (or equivalent) in a related area/subject.

Eligibility

Applicants must have obtained or be about to obtain a minimum Upper Second class UK honours degree, or the equivalent qualifications gained outside the UK, in a relevant discipline.

Before you Apply

Applicants must make direct contact with preferred supervisors before applying. It is your responsibility to make arrangements to meet with potential supervisors, prior to submitting a formal online application.

How to Apply

To be considered for this project you MUST submit a formal online application form – on the application form select PhD Cardiovascular Sciences Programme. Full details on how to apply can be found on the Website: How to apply for postgraduate research at The University of Manchester

If you have any queries regarding making an application please contact our admissions team FBMH.doctoralacademy.admissions@manchester.ac.uk

Funding Notes

Applications are invited from self-funded students. This project has a Band 3 (high) fee. Details of our different fee bands can be found on our website View Website

References

Bhuiyan ZA, van den Berg MP, van Tintelen JP et al. Expanding spectrum of human RYR2-related disease: new electrocardiographic, structural, and genetic features Circulation. 2007 Oct 2;116(14):1569-76
Steinberg C, Roston TM, van der Werf C et al. RYR2-ryanodinopathies: from calcium overload to calcium deficiency Europace. 2023 Jun 2;25(6):euad156
Toth N, Zhang XH, Zamaro A, Morad M. Calcium Signaling Consequences of RyR2-S4938F Mutation Expressed in Human iPSC-Derived Cardiomyocytes Int J Mol Sci. 2023 Oct 18;24(20):15307