Scientific basis: Heart failure (HF) affects 64 million people globally and causes sudden death. Cardiac fibrosis (CF) drives HF creating a need to image CF and therapeutically target it. Current imaging technologies do not directly measure CF and no specific antifibrotic drugs are available. Secreted matricellular proteins have been shown to inhibit CF and preserve cardiac contractility in vivo by inhibiting TGFβ signalling. An engineered effector T cell immunotherapy targeting myofibroblasts was reported to treat cardiac fibrosis in mice. Moreover, regulatory T cell (Treg) therapy has been proposed as a candidate to counteract cardiac inflammation- induced CF. But, cell therapy homing, survival and function of the cells at the target sites remain elusive. To develop advanced methods to diagnose CF and quantify cell therapy distribution in the body, this project will employ non-invasive in vivo imaging for cell tracking (Fruhwirth) alongside direct quantification of fibrosis (Phinikaridou) to monitor the effectiveness of novel anti-CF therapies.
Aims & objectives for each year
(1) Establish an in vivo animal model of CF and use fibrosis-specific MRI-probes to quantify the therapeutic window (Year 1).
(2) Engineer and in vitro characterise Tregs to release a protein with proven activity against CF (AF-Tregs) (Year 1).
(3) Administer AF-Tregs in vivo and quantify homing and survival in healthy and diseased hearts by molecular PET to detect the cell therapy (Year 2).
(4) Employ PET/MRI imaging to quantify AF-Treg efficacy alongside monitoring of disease status/treatment outcome using the fibrosis-specific MRI-probes (Year 2.5 to 3.5)
o Immune cell isolation, purification, and culture (focus on Tregs)
o Treg engineering (to secret an anti-fibrotic protein and be traceable in vivo)
o Characterise Tregs (flow cytometry and imaging reporter)
o Mouse husbandry, handling, and surgical techniques.
o In vivo PET and cardiac MRI.
o Ex vivo histology, proteomics, western blotting, ELISA.