Reilly Group: Adverse myocardial remodelling in cardiac fibrosis and atrial fibrillation.

Our main focus is to understand the mechanisms of two major often linked heart conditions cardiac fibrosis and atrial fibrillation.

Confocal microscopy image showing human atrial fibroblast co-stained for actin-binding protein filamin A and GPCR calcitonin receptor.

About the Research

The main goal of our research is to uncover new important mechanisms underlying and causing these challenging conditions in order to identify new drug-targets. To achieve our research goal we utilise a combination of extensive cell culture, molecular, RNA/DNA/microRNA biology techniques, functional cellular assays and advanced high throughput approaches performed in human cardiac and blood samples, iPSC-derived cardiomyocytes, and, to a lesser extent, animal models (e.g., mice and guinea pigs). The work involving clinical human cardiac and blood samples may be carried out by a student with or without GMC registration. Animals licence is not required for these studies.

Cardiac fibrosis is a hallmark histological feature of structural changes in the myocardium associated with virtually all cardiac diseases (e.g., heart failure, hypertension, and atrial fibrillation). To date, there is no effective treatment for cardiac fibrosis, as we do not understand the mechanisms contributing, or causing it. Our group is very interested in uncovering new potentially important pathways accountable for this condition. Specifically, we explore the role of G-protein coupled receptors (GPCRs), e.g., calcitonin, amylin and CGRP receptors, and their downstream signalling pathways involved in cardiac fibrogenesis [1]. As part of this theme we are also looking at the role of RAMPs (Receptor activity modifying proteins) in regulation of GPCR trafficking. In this project, we also aim to comprehensively study disease-specific transcriptional and proteome signatures of cardiac tissue with a focus on fibroblasts subtypes and origine using single-cell RNA-sequencing and spatical proteome approaches [1]. These studies will help us to uncover new important mechanisms causing and contributing to the development of cardiac fibrosis, a progressive and currently incurrable condition.

Atrial fibrillation (AF) is the most common cardiac rhythm disorder, called arrhythmia, in humans. Changes in structural and electrical properties of the heart have been long implicated in this arrhythmia. However, the upstream mechanisms underlying changes in both structural (cardiac fibrosis) and electrical myocardial changes in atrial fibrillation are still unclear. Thus, we are interested in elucidating functional and structural responses of human (to a lesser extent, murine) myocardium and cardiac cells (i.e., cardiomyocytes and fibroblasts), to a number of singalining molecules, including but not limited to calcitonin, CGRP [1], amylin, interleukin-11 and interleukin-6. Our lab also explores changes in structural proteins such as filamin A and their therapeutic potenial in AF.

Clinical work relevant to this project will be performed in patients’ blood and cardiac tissue/cell samples collected by the student in close collaboration with NHS teams of clinicians, cardiothoracic surgeons and EP teams based at Oxford Heart Centre (John Radcliffe hospital).

Animal studies would be limited in this pojrect but will use sophisticated genetically modified mouse models of AF [2] with a gene deletion or overexpression targeted to a specific heart chamber (i.e., atria or ventricle) and cell type (i.e., fibroblasts and myocytes) [1]. This part of projects will be performed in collaboration with the top international labs (in the USA and Canada). Some animal work (guinea pigs) is done in collaboration with the local CVMed collaborators (e.g., Prof Charles Redwood and Dr Paul Robinson).

Biomarkers and Drug-discovery studies (libraries screens) will be done in collaboration with TDI, Oxford, and industry.

High throughput work complements all above studies and are carried out in collaboration with Dusseldorf Institute of Immunology, Azenta (Germany) and BIogenity (Denmark) labs.

All our work is faciliateted by a number of internal and external collaborations including TDI (Oxford), CVMed (Oxford), Imperial College London, Montreal Heart Institute (Canada), Baylor College of Medicine (USA), Essen Institute of Pharmacology (Germany), and Dusseldorf Institute of Immunology (Germany).

Potential student would have an opportunity to work with a team of enthusiastic, hard working and friendly scientists on the outlined above themes with access to a wide range of RNA/molecular and cellular biology techniques (see section “Training opporunities”). There will be a great opportunity to work in collaborating UK-based (e.g., Oxford University, King’s College, and Manchester University) and international labs (Montreal Heart Institute and Baylor College of Medicine) and acquire some fundamental techniques in fibroblast and cardiomyocyte function, sn/sc/bulk-RNA sequencing, spacial proteomics, and designing new animal models. The student would also have an opportunity to learn how to recruit and consent patients for clinical studies, clinical data mining and how to process human blood samples, human heart biopsies and/or (when required for the project) work with animal models (e.g., mice and guinea pigs).

Training Opportunities 

We offer training in the following techniques relevant to the ongoing projects:

• Molecular biology, including (but not limited to) immunoblotting, immunostaining, ELISA, cloning, RNA/DNA extraction, qPCR, PCR, Trichrome Masson’s staining.
• Extensive cell culture techniques in primary human and rodent fibroblasts and myocytes, or in cell lines (e.g., HEK293 and 3T3).
• Cellular functional studies (including assessment of cell viability, proliferation, migration and wound healing; loss-of- and gain-of-function studies using lipo/electroporation transfection protocols with siRNA/vectors.
• Clinical studies will involve patients’’ recruitment and consenting, clinical datamining, collection of human blood sample and cardiac biopsies for a subsequent measurement of biomarkers in atrial fibrillation and studies into cardiac fibrosis.
• Assessment of calcium handling (e.g., contractility, cell relaxation, calcium transients) and electrophysiology of cardiomyocytes as a part of internal and international ongoing collaborations.
• Animal work – only when required by the project - (mice and guinea pigs) including breeding, colony maintenance, assessment of cardiac fibrosis and arrhythmogenesis in vivo and in vitro.
• Some training in RNA-sequencing (including single cell) and spatial proteomics maybe offered dependent on the project.
• iPSC work (in collaboration with local experts).

Students are encouraged to attend the MRC Weatherall Institute of Molecular Medicine DPhil Course, which takes place in the autumn of their first year. Running over several days, this course helps students to develop basic research and presentation skills, as well as introducing them to a wide range of scientific techniques and principles, ensuring that students have the opportunity to build a broad-based understanding of differing research methodologies.

Generic skills training is offered through the Medical Sciences Division's Skills Training Programme. This programme offers a comprehensive range of courses covering many important areas of researcher development: knowledge and intellectual abilities, personal effectiveness, research governance and organisation, and engagement, influence, and impact. Students are actively encouraged to take advantage of the training opportunities available to them.

As well as the specific training detailed above, students will have access to a wide range of seminars and training opportunities through the many research institutes and centres based in Oxford.

The Department has a successful mentoring scheme, open to graduate students, which provides an additional possible channel for personal and professional development outside the regular supervisory framework. We hold an Athena SWAN Silver Award in recognition of our efforts to build a happy and rewarding environment where all staff and students are supported to achieve their full potential.

Publications

Publications: