(MRC DTP) STORM and CLEM Imaging to Understand the Pathophysiology of the Failing Heart

This project will use state-of-the art imaging techniques to define how the organisation of cardiac cellular calcium handling proteins is perturbed in the setting of cardiovascular disease. These imaging techniques include Stochastic Optical Reconstruction Microscopy (STORM) and Correlative Light and Electron Microscopy (CLEM) techniques [1]. The project builds upon the established track record of the supervisory teams laboratory in defining how intracellular calcium handling controls cardiac function and how perturbations in this system lead to contractile dysfunction, arrhythmias and impaired responsiveness to catecholamines [e.g. 2-4] and application of electron microscopy approaches to define alterations to cardiac cellular structure in heart failure [5]. The main question that the project seeks to address is that of the role that altered positioning of the components of the cardiac dyad relative to the transverse tubule membrane plays in the contractile dysfunction, increased propensity for arrhythmias and, impaired responsiveness to adrenergic (catecholamine) stimulation in heart failure. To achieve this aims the student will join a large and vibrant group and receive training in the in vivo methodologies required to deliver a translationally relevant model of heart failure [e.g. 2-5]. Tissues and single viable isolated cardiac myocytes will be collected from this model and cellular calcium regulation assessed using calcium sensitive fluorescent dyes and confocal microscopy. These samples will then be processed for immunohistological and electron microscopic assessment of cardiac dyadic organisation and composition. An example of the type of outcomes this project will deliver is a definitive assessment of the size, distribution and homogeneity of ryanodine receptor clusters in heart failure. This structural information will help, for example, elucidate how changes in the nano-organisation of the ryanodine receptor cluster size; i) aspects of contractile dysfunction such as impaired release of calcium and reduced contraction and, ii) identify likely triggers for arrhythmias in the form of the probability of and location of calcium sparks (as measured by confocal microscopy with calcium sensitive dyes). Ultimately, by identifying critical aspects of maladaptive structural remodelling in heart failure it is anticipated that novel treatment regimes can be devised.
Personal Pages

https://www.research.manchester.ac.uk/portal/Andrew.W.Trafford.html

https://www.research.manchester.ac.uk/portal/Katharine.Dibb.html

Our Work highlighted by British Heart Foundation

https://www.bhf.org.uk/heart-matters-magazine/research/research-projects-on-heart-failure