H2020 MSCA-IF 2021-2023


Marie Skłodowska-Curie Individual Fellowship to Dr Monika Biniecka under supervision of Professor Józef Dulak


„Mechanistic links between rheumatoid arthritis and cardiovascular complications: investigation on inflammation induced alterations in induced pluripotent stem cell-derived cardiomyocytes”, acronym CVD in RA, ID no: 841627

 

Rheumatoid arthritis (RA) is a chronic and progressive inflammatory disease of the joints, characterized by synovial inflammation and destruction of cartilage and bone. Nowadays, RA is considered as a syndrome that also includes systemic inflammation, extra-articular manifestations and several comorbidities. In particular, the risk of cardiovascular disease (CVD) in RA patients is 1.5–2-fold higher compared the general population due to the persistent systemic inflammation leading to increased damage to cardiac and endothelial cells. Although this recognition has been described by clinicians, more studies are needed to identify CVD risk factors which may be specific to RA patients, including medications used to manage RA. For this reason, this project aims to generate a novel cellular tool that may help to uncover the molecular mechanisms by which RA patients may have an elevated risk of CV complications.

Human heart cells are difficult to access from the heart of RA patients. Therefore, in this project, we will implement the technology of induced pluripotent stem cells (iPSCs), pioneered by Shinya Yamanaka’s lab in Kyoto, Japan, who was awarded the 2012 Nobel Prize. iPSCs are a type of stem cell that can be generated directly from adult cells like fibroblasts or blood cells. iPSCs have the same properties as embryonic stem cells, i.e., self-renewing and pluripotent differentiation giving rise to many other cell types, such as neurons, heart, pancreatic, and liver cells. In this project we will employ a truly translational, bench-to-bedside approach build on the strength of the human RA model. In collaboration with Rheumatologists, human samples of joint fibroblasts (FLS) and peripheral blood mononuclear cells (PBMC) will be collected from RA patients. A reprogramming method with Yamanaka cocktail will allow us to generate FLS-derived iPSCs and PBMC-derived iPSCs. Next, using the novel differentiation strategy, iPSCs will be converted into the cardiomyocytes (CMs). To imitate the impact of the joint inflammation and medications on cardiomyocytes properties and functions, cells will be stimulated with key RA pro-inflammatory cytokines and anti-RA drugs. Later, the state-of-the-art technologies will be applied, such immunofluorescence microscopy, flow cytometry, patch-clamp technology, cell metabolism analyzer Seahorse and polymerase chain reaction (PCR).

Applying the above techniques, we will firstly confirm if established in this project iPSCs and CMs are functional and possess the desire cell-specific markers and properties. Secondly, we will investigate if RA pro-inflammatory cytokines and anti-RA drugs affect the cardiac cells functions including electrophysiology and bioenergetics.

Overall, we expect that the results of project will allow to better understand the molecular mechanism leading to the CV complications in RA and whether the application of iPSC-derived cardiac cells can be a useful platform for evaluation drug toxicity.